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Sample records for induced fluorescence method

  1. A Method to Reconstruct the Solar-Induced Canopy Fluorescence Spectrum from Hyperspectral Measurements

    Directory of Open Access Journals (Sweden)

    Feng Zhao

    2014-10-01

    Full Text Available A method for canopy Fluorescence Spectrum Reconstruction (FSR is proposed in this study, which can be used to retrieve the solar-induced canopy fluorescence spectrum over the whole chlorophyll fluorescence emission region from 640–850 nm. Firstly, the radiance of the solar-induced chlorophyll fluorescence (Fs at five absorption lines of the solar spectrum was retrieved by a Spectral Fitting Method (SFM. The Singular Vector Decomposition (SVD technique was then used to extract three basis spectra from a training dataset simulated by the model SCOPE (Soil Canopy Observation, Photochemistry and Energy fluxes. Finally, these basis spectra were linearly combined to reconstruct the Fs spectrum, and the coefficients of them were determined by Weighted Linear Least Squares (WLLS fitting with the five retrieved Fs values. Results for simulated datasets indicate that the FSR method could accurately reconstruct the Fs spectra from hyperspectral measurements acquired by instruments of high Spectral Resolution (SR and Signal to Noise Ratio (SNR. The FSR method was also applied to an experimental dataset acquired in a diurnal experiment. The diurnal change of the reconstructed Fs spectra shows that the Fs radiance around noon was higher than that in the morning and afternoon, which is consistent with former studies. Finally, the potential and limitations of this method are discussed.

  2. Laser-induced fluorescence as a method of early caries diagnosis

    Science.gov (United States)

    Mielczarek, Agnieszka; Wiewior, Piotr

    2001-07-01

    Use of lasers in dentistry dates back 20 years but is still not fully exploited, especially when concerning the hard dental tissues. Over the past many efforts and actions have been involved in testing and developing new methods for caries diagnosis. The implementation of these methods in general dental practice is unfortunately still limited because too little scientific evidence exists to support them. One of the age-old concerns for dentists is that decay is often discovered too late. Dentists commonly use x-ray imaging for early caries detection, but this method cannot reveal decay at a sufficiently early stage to avoid restorative methods. Generally, if a caries lesion si detected by x-ray, the mineral loss within the tooth is normally very high and will need invasive treatment. Several laser based techniques, as also other optical methods of detecting caries lesions at an early stage seem to be very promising. Fluorescence of tooth structure is observed when hard tissues are illuminated using laser light. Decayed areas appear dark and provide a contrast against the healthy background surrounding them, so discriminating sound and carious tissues. The aim of this study was to present the possibilities of using laser induced fluorescence in the diagnosis of early caries lesions. The current state-of-the-art is presented, as well as results of our investigations. In our studies an argon ion laser was used to illuminate the teeth and the fluorescence pictures were captured with a CCD camera and then analyzed. Results confirmed that laser induced fluorescence can be used as a sensitive method of caries diagnosis.

  3. Diagnostics of Susabi-nori (Porphyra Yezoensis) by Laser-Induced Fluorescence Method

    Science.gov (United States)

    Okamoto, Tamotsu; Nakamura, Yuki; Takahashi, Kunio; Kaneko, Shohei; Shimada, Yuji

    Susabi-nori (Porphyra yezoensis) was diagnosed by means of laser-induced fluorescence (LIF) method. Fluorescence peaks located at approximately 580, 660, 685 and 720 nm were observed in the LIF spectra of Susabi-nori. In the spectrum of the sample infected with the red rot disease, the intensity of 580 nm peak was relatively high as compared with that of the control sample. On the other hand, the intensities of 580 nm and 660 nm peaks drastically decreased by the influence of the chytrid disease. Furthermore, the intensity of the 580 nm peak increased by dipping into fresh water. These results indicate that LIF spectra of Susabi-nori are affected by the diseases and the stress of fresh water and that the diseases and the stress of Susabi-nori can be diagnosed by the LIF method.

  4. Field Observations with Laser-Induced Fluorescence Transient (LIFT Method in Barley and Sugar Beet

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    Anna R. Raesch

    2014-05-01

    Full Text Available The laser-induced fluorescence transient (LIFT method is a non-invasive remote sensing technique for measurement of photosynthetic performance of plants under laboratory and field conditions. We report here a long-term comparative study to monitor the performance of different cultivars of barley and sugar beet during the growth season of these crops. The LIFT measurements provided useful results about photosynthetic light use efficiency on selected leaves in the canopy of the studied crops. The different canopy architectures, with different optical properties, influenced the LIFT measurements.

  5. Experimental Research of Reliability of Plant Stress State Detection by Laser-Induced Fluorescence Method

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    Yury Fedotov

    2016-01-01

    Full Text Available Experimental laboratory investigations of the laser-induced fluorescence spectra of watercress and lawn grass were conducted. The fluorescence spectra were excited by YAG:Nd laser emitting at 532 nm. It was established that the influence of stress caused by mechanical damage, overwatering, and soil pollution is manifested in changes of the spectra shapes. The mean values and confidence intervals for the ratio of two fluorescence maxima near 685 and 740 nm were estimated. It is presented that the fluorescence ratio could be considered a reliable characteristic of plant stress state.

  6. Experimentally studied laser fluorescence method for remote sensing of plant stress situation induced by improper plants watering

    Directory of Open Access Journals (Sweden)

    Yu. V. Fedotov

    2014-01-01

    Full Text Available Stressful situations of plants can be caused by a lack of nutrients; mechanical damages; diseases; low or high temperatures; lack of illumination; insufficient or excess humidity of the soil; soil salinization; soil pollution by oil products or heavy metals; the increased acidity of the soil; use of pesticides, herbicides, insecticides, etc.At early stages it is often difficult to detect seemingly that the plants are in stressful situations caused by adverse external factors. However, the fluorescent analysis potentially allows detection of the stressful situations of plants by deformation of laser-induced fluorescence spectra. The paper conducts experimental investigations to learn the capabilities of the laser fluorescent method to monitor plant situations at 532nm wavelength of fluorescence excitation in the stressful situations induced by improper watering (at excess of moisture in the soil and at a lack of moisture.Researches of fluorescence spectra have been conducted using a created laboratory installation. As a source to excite fluorescence radiation the second harmonica of YAG:Nd laser is used. The subsystem to record fluorescence radiation is designed using a polychromator and a highly sensitive matrix detector with the amplifier of brightness.Experimental investigations have been conducted for fast-growing and unpretentious species of plants, namely different sorts of salad.Experimental studies of laser-induced fluorescence spectra of plants for 532nm excitement wavelength show that the impact of stressful factors on a plant due to the improper watering, significantly distorts a fluorescence spectrum of plants. Influence of a stressful factor can be shown as a changing profile of a fluorescence spectrum (an identifying factor, here, is a relationship of fluorescence intensities at two wavelengths, namely 685 nm and 740 nm or (and as a changing level of fluorescence that can be the basis for the laser method for monitoring the plant

  7. A method for the measurement of in line pistachio aflatoxin concentration based on the laser induced fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Paghaleh, Soodeh Jamali [Vali-e-Asr University of Rafsanjan, Rafsanjan (Iran, Islamic Republic of); Askari, Hassan Ranjbar; Marashi, Seyed Mohammad Bagher [Department of Physics, Faculty of Science, Vali-e-Asr University of Rafsanjan, Rafsanjan (Iran, Islamic Republic of); Rahimi, Mojtaba, E-mail: m_rahimi@vru.ac.ir [Department of Physics, Faculty of Science, Vali-e-Asr University of Rafsanjan, Rafsanjan (Iran, Islamic Republic of); Bahrampour, Ali Reza [Physics Department of Sharif University of Technology, Tehran (Iran, Islamic Republic of)

    2015-05-15

    Contamination of pistachio nuts with aflatoxin is one of the most significant issues related to pistachio health and expert. A fast pistachio aflatoxin concentration measurement method based on the laser induced fluorescence spectroscopy (LIFS) is proposed. The proposed method from theoretical and experimental points of view is analyzed. In our experiments XeCl Excimer laser is employed as an Ultra Violet (UV) source (λ=308 nm) and a UV–visible (UV–vis) spectrometer is used for fluorescent emission detection. Our setup is employed to measure the concentration of different type of Aflatoxins in pistachio nuts. Measurements results obtained by the LIFS method are compared with those are measured by the standard HPLC method. Aflatoxins concentrations are in good agreement with those are obtained by the HPLC method. The proposed laser induced fluorescence spectroscopy can be used as an in line aflatoxins concentrations measurement instrument for industrial applications. - Highlights: • XeCl Excimer laser is employed as an UV source for measurement of AFs in pistachio nuts. • Results are compared with those are measured by the standard HPLC method. • LIFS is an online AFs concentration measurement method for industrial applications.

  8. Fluorescence-based methods for the detection of pressure-induced spore germination and inactivation

    Science.gov (United States)

    Baier, Daniel; Reineke, Kai; Doehner, Isabel; Mathys, Alexander; Knorr, Dietrich

    2011-03-01

    The application of high pressure (HP) provides an opportunity for the non-thermal preservation of high-quality foods, whereas highly resistant bacterial endospores play an important role. It is known that the germination of spores can be initiated by the application of HP. Moreover, the resistance properties of spores are highly dependent on their physiological states, which are passed through during the germination. To distinguish between different physiological states and to detect the amount of germinated spores after HP treatments, two fluorescence-based methods were applied. A flow cytometric method using a double staining with SYTO 16 as an indicator for germination and propidium iodide as an indicator for membrane damage was used to detect different physiological states of the spores. During the first step of germination, the spore-specific dipicolinic acid (DPA) is released [P. Setlow, Spore germination, Curr. Opin. Microbiol. 6 (2003), pp. 550-556]. DPA reacts with added terbium to form a distinctive fluorescent complex. After measuring the fluorescence intensity at 270 nm excitation wavelength in a fluorescence spectrophotometer, the amount of germinated spores can be determined. Spores of Bacillus subtilis were treated at pressures from 150 to 600 MPa and temperatures from 37 °C to 60 °C in 0.05 M ACES buffer solution (pH 7) for dwell times of up to 2 h. During the HP treatments, inactivation up to 2log 10 cycles and thermal sensitive populations up to 4log 10 cycles could be detected by plate counts. With an increasing number of thermal sensitive spores, an increased proportion of spores in germinated states was detected by flow cytometry. Also the released amount of DPA increased during the dwell times. Moreover, a clear pressure-temperature-time-dependency was shown by screening different conditions. The fluorescence-based measurement of the released DPA can provide the opportunity of an online monitoring of the germination of spores under HP inside

  9. Remote monitoring of 129I and 127I isotopes in the atmosphere using the laser-induced fluorescence method

    Science.gov (United States)

    Kireev, S. V.; Shnyrev, S. L.; Suganeev, S. V.

    2016-09-01

    This paper reports the experimental and calculation research of the development of the remote laser-induced fluorescence method for the detection of 129I and 127I molecular iodine isotopologues in atmospheric air in real time. As an excitation source we used a frequency-doubled neodymium laser (~532 nm). We estimated the sensitivity of 127I129I and 129I2 detection in the atmosphere. Detection sensitivity of molecular iodine is 4 · 1013 cm-3 for a sensing distance of 6 km.

  10. Fluorescence nanoscopy. Methods and applications

    OpenAIRE

    Requejo-Isidro, Jose

    2013-01-01

    Fluorescence nanoscopy refers to the experimental techniques and analytical methods used for fluorescence imaging at a resolution higher than conventional, diffraction-limited, microscopy. This review explains the concepts behind fluorescence nanoscopy and focuses on the latest and promising developments in acquisition techniques, labelling strategies to obtain highly detailed super-resolved images and in the quantitative methods to extract meaningful information from them.

  11. Comparison of light-induced and laser-induced fluorescence methods for the detection and quantification of enamel demineralization

    Science.gov (United States)

    Ando, Masatoshi; Analoui, Mostafa; Schemehorn, Bruce R.; Stookey, George K.

    1999-05-01

    The Quantitative Laser-Induced Fluorescence (QLF) technique has been sued for diagnosis of early caries in permanent teeth (PT). The objective of this study was to determine the caries quantification ability of QLF in deciduous teeth (DT). Sixty sound teeth, thirty DT and thirty PT, were used. All teeth were cleaned to remove debris and equally divided into three groups. Lesions were created in small windows (0.8x2.0 mm2) on buccal or labial surface for 48, 72, and 96 hr. Lesion images were made with a 488 nm argon laser (QLF I) and then with a 370 +/- 80 nm violet-blue light (QLF II). Both images were analyzed to determine the mean percent change in fluorescence radiance (ΔF). A center section from the lesions was taken for analysis with microradiography. The lesion depth and loss of mineral content were determined. The correlations between ΔF and lesion depth as well as ΔZ in DT were 0.76 and 0.84 with QLF I, 0.81 and 0.88 with QLF II, respectively. It can be concluded the ability of QLF to quantify white-spots in DT is better than in PT.

  12. A conformation-induced fluorescence method for microRNA detection

    DEFF Research Database (Denmark)

    Aw, Sherry S; Tang, Melissa Xm; Teo, Yin Nah

    2016-01-01

    MicroRNAs play important roles in a large variety of biological systems and processes through their regulation of target mRNA expression, and show promise as clinical biomarkers. However, their small size presents challenges for tagging or direct detection. Innovation in techniques to sense...... and quantify microRNAs may aid research into novel aspects of microRNA biology and contribute to the development of diagnostics. By introducing an additional stem loop into the fluorescent RNA Spinach and altering its 3' and 5' ends, we have generated a new RNA, Pandan, that functions as the basis for a microRNA...... sensor. Pandan contains two sequence-variable stem loops that encode complementary sequence for a target microRNA of interest. In its sensor form, it requires the binding of a target microRNA in order to reconstitute the RNA scaffold for fluorophore binding and fluorescence. Binding of the target microRNA...

  13. A sensitive fluorescence anisotropy method for detection of lead (II) ion by a G-quadruplex-inducible DNA aptamer

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Dapeng [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085 (China); Yin, Lei; Meng, Zihui [School of Chemical Engineering and Environment, Beijing Institute of Technology, Beijing, 100081 (China); Yu, Anchi [Department of Chemistry, Renmin University of China, Beijing, 100872 (China); Guo, Lianghong [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085 (China); Wang, Hailin, E-mail: hlwang@rcees.ac.cn [State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences, Beijing, 100085 (China)

    2014-02-17

    Graphical abstract: -- Highlights: •A fluorescence anisotropy approach for detection of Pb{sup 2+} was developed. •The strategy was based on binding-induced allosteric conformational change of aptamer probe. •The sensing mechanism was established by testing the photoinduced electron transfer interaction. -- Abstract: Sensitive and selective detection of Pb{sup 2+} is of great importance to both human health and environmental protection. Here we propose a novel fluorescence anisotropy (FA) approach for sensing Pb{sup 2+} in homogeneous solution by a G-rich thrombin binding aptamer (TBA). The TBA labeled with 6-carboxytetramethylrhodamine (TMR) at the seventh thymine nucleotide was used as a fluorescent probe for signaling Pb{sup 2+}. It was found that the aptamer probe had a high FA in the absence of Pb{sup 2+}. This is because the rotation of TMR is restricted by intramolecular interaction with the adjacent guanine bases, which results in photoinduced electron transfer (PET). When the aptamer probe binds to Pb{sup 2+} to form G-quadruplex, the intramolecular interaction should be eliminated, resulting in faster rotation of the fluorophore TMR in solution. Therefore, FA of aptamer probe is expected to decrease significantly upon binding to Pb{sup 2+}. Indeed, we observed a decrease in FA of aptamer probe upon Pb{sup 2+} binding. Circular dichroism, fluorescence spectra, and fluorescence lifetime measurement were used to verify the reliability and reasonability of the sensing mechanism. By monitoring the FA change of the aptamer probe, we were able to real-time detect binding between the TBA probe and Pb{sup 2+}. Moreover, the aptamer probe was exploited as a recognition element for quantification of Pb{sup 2+} in homogeneous solution. The change in FA showed a linear response to Pb{sup 2+} from 10 nM to 2.0 μM, with 1.0 nM limit of detection. In addition, this sensing system exhibited good selectivity for Pb{sup 2+} over other metal ions. The method is simple

  14. The evaluation of a novel method comparing quantitative light-induced fluorescence (QLF) with spectrophotometry to assess staining and bleaching of teeth

    NARCIS (Netherlands)

    Adeyemi, A.A.; Jarad, F.D.; de Josselin de Jong, E.; Pender, N.; Higham, S.M.

    2010-01-01

    This study reports the development and evaluation of a novel method using quantitative light-induced fluorescence (QLF), which enables its use for quantifying and assessing whole tooth surface staining and tooth whitening. The method was compared with a spectrophotometer to assess reliability. Two

  15. The evaluation of a novel method comparing quantitative light-induced fluorescence (QLF) with spectrophotometry to assess staining and bleaching of teeth

    NARCIS (Netherlands)

    Adeyemi, A.A.; Jarad, F.D.; de Josselin de Jong, E.; Pender, N.; Higham, S.M.

    2010-01-01

    This study reports the development and evaluation of a novel method using quantitative light-induced fluorescence (QLF), which enables its use for quantifying and assessing whole tooth surface staining and tooth whitening. The method was compared with a spectrophotometer to assess reliability. Two e

  16. The evaluation of a novel method comparing quantitative light-induced fluorescence (QLF) with spectrophotometry to assess staining and bleaching of teeth

    NARCIS (Netherlands)

    Adeyemi, A.A.; Jarad, F.D.; de Josselin de Jong, E.; Pender, N.; Higham, S.M.

    2010-01-01

    This study reports the development and evaluation of a novel method using quantitative light-induced fluorescence (QLF), which enables its use for quantifying and assessing whole tooth surface staining and tooth whitening. The method was compared with a spectrophotometer to assess reliability. Two e

  17. Laser-induced fluorescence imaging of bacteria

    Science.gov (United States)

    Hilton, Peter J.

    1998-12-01

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

  18. Protein Profile study of clinical samples using Laser Induced Fluorescence as the detection method

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Raja, Sujatha N.; Rai, Lavanya;

    2009-01-01

      Protein profiles of tissue homogenates were recorded using HPLC separation and LIF detection method. The samples were collected from volunteers with clinically normal or cervical cancer conditions. It is shown that the protein profile can be classified as belonging to malignant or normal state ...

  19. New Spectral Fitting Method for Full-Spectrum Solar-Induced Chlorophyll Fluorescence Retrieval Based on Principal Components Analysis

    Directory of Open Access Journals (Sweden)

    Xinjie Liu

    2015-08-01

    Full Text Available The full-spectrum Solar-Induced chlorophyll Fluorescence (SIF within the 650-800 nm spectral region can provide important information regarding physiological and biochemical activities in vegetation. This paper proposes a new Full-spectrum Spectral Fitting Method (F-SFM for the retrieval of SIF spectra based on Principal Components Analysis (PCA. Using F-SFM, both the full-spectrum reflectance and SIF within the 650-800 nm region were modeled by PCA based on a training dataset simulated by the Soil Canopy Observation, Photochemistry and Energy fluxes (SCOPE model, and the weighting coefficients of the principal components were estimated by the least-squares fitting method. An iterative process was employed to improve the accuracy of the estimation of the reflectance. In each iteration, the SIF spectra retrieved from the last run were removed from the total upwelling radiance to minimize the small contribution of the SIF to the apparent reflectance outside the absorption bands. Then, the F-SFM algorithm was tested using both simulated and field-measured data with different Spectral Resolutions (SRs and Signal-to-Noise Ratios (SNRs. For data with an SR of 0.3 nm and without noise, the Relative Root Mean Square Error (RRMSE was less than 14% within the spectral region that was studied, and the peak-value ratio (SIF735/SIF685 was accurately estimated with an RRMSE of 3.56%. In addition, the F-SFM algorithm proved less sensitive to the SR than the three-band Fraunhofer Line Discrimination (3 FLD and improved FLD (iFLD methods. In the case of the field spectral data with SRs of 3 nm and 0.3 nm, the double-peak shape and the diurnal variation trend of the SIF spectra could be reasonably reconstructed by F-SFM, and the retrieved SIF values at the O2-A and O2-B bands were consistent with those retrieved by 3FLD from data with a high SR (0.3 nm and SNR (1000. Therefore, the F-SFM method can provide full-spectrum SIF information with high accuracy even at

  20. Detection of acrylamide in potato chips using a fluorescent sensing method based on acrylamide polymerization-induced distance increase between quantum dots.

    Science.gov (United States)

    Hu, Qinqin; Xu, Xiahong; Li, Zhanming; Zhang, Ying; Wang, Jianping; Fu, Yingchun; Li, Yanbin

    2014-04-15

    Acrylamide is a neurotoxin and potential carcinogen, but is found in various thermally processed foods such as potato chips, biscuits, and coffee. Simple and sensitive methods for on-line detection of acrylamide are needed to ensure food safety. In this paper, a novel fluorescent sensing method based on acrylamide polymerization-induced distance increase between quantum dots (QDs) was proposed for detecting acrylamide in potato chips. The functional QDs were prepared by their binding with N-acryloxysuccinimide (NAS), which was characterized by Fourier transform infrared (FR-IR) spectra. The carbon-carbon double bonds of NAS modified QDs polymerized with assistance of photo initiator under UV irradiation, leading to QDs getting closer along with fluorescence intensity decreasing. Acrylamide in the sample participated in the polymerization and induced an increase of fluorescence intensity. This method possessed a linear range from 3.5×10(-5) to 3.5 g L(-1) (r(2)=0.94) and a limit of detection of 3.5×10(-5) g L(-1). Although the sensitivity and specificity cannot be compared with standard LC-MS/MS analysis, this new method requires much less time and cost, which is promising for on-line rapid detection of acrylamide in food processing.

  1. A new direct laser photo-induced fluorescence method coupled on-line with liquid chromatographic separation for the simultaneous determination of anilides pesticides.

    Science.gov (United States)

    Mbaye, O M A; Maroto, A; Gaye-Seye, M D; Stephan, L; Deschamps, L; Aaron, J J; Giamarchi, P

    2015-01-01

    A new direct laser photo-induced fluorescence high performance liquid chromatography (DL-PIF-HPLC) method is developed for the simultaneous determination of three anilide pesticides, namely carboxin, monalide and propanil. DL-PIF-HPLC uses a tunable Nd:YAG-OPO laser to obtain fluorescent photoproduct(s) and to simultaneously analyze their fluorescence in a short acquisition time with an intensified CCD camera, which improves the selectivity (by choosing the suitable excitation wavelength), increases the sensitivity (due to the high energy of the laser beam) and reduces the time of analysis, relative to the classical PIF methods. However, one of the main drawbacks of PIF methods is the presence of interferences with other compounds, such as other pesticides from the same group yielding similar fluorescent photoproducts, which reduces their selectivity. The analytical interest of DL-PIF-HPLC to avoid these interferences is demonstrated. The DL-PIF spectra, chromatographic conditions and analytical performances of DL-PIF-HPLC are presented for the simultaneous determination of three anilide pesticides. The calibration curves are linear over one order of magnitude and the limits of detection are in the ng mL(-1) range. The new DL-PIF-HPLC system has the advantage to combine the performances of both techniques, DL-PIF and liquid chromatography, and to improve the analysis selectivity.

  2. Fluorescence calibration method for single-particle aerosol fluorescence instruments

    Science.gov (United States)

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

    2017-05-01

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

  3. Direct methods for dynamic monitoring of secretions from single cells by capillary electrophoresis and microscopy with laser-induced native fluorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Tong, W.

    1997-10-08

    Microscale separation and detection methods for real-time monitoring of dynamic cellular processes (e.g., secretion) by capillary electrophoresis (CE) and microscopic imaging were developed. Ultraviolet laser-induced native fluorescence (LINF) provides simple, sensitive and direct detection of neurotransmitters and proteins without any derivatization. An on-column CE-LINF protocol for quantification of the release from single cell was demonstrated. Quantitative measurements of both the amount of insulin released from and the amount remaining in the cell ({beta}TC3) were achieved simultaneously. Secretion of catecholamines (norepinephrine (NE) and epinephrine (E)) from individual bovine adrenal chromaffin cells was determined using the on-column CE-LINF. Direct visualization of the secretion process of individual bovine adrenal chromaffin cells was achieved by LINF imaging microscopy with high temporal and spatial resolution. The secretion of serotonin from individual leech Retzius neurons was directly characterized by LINF microscopy with high spatial resolution.

  4. Direct methods for dynamic monitoring of secretions from single cells by capillary electrophoresis and microscopy with laser-induced native fluorescence detection

    Energy Technology Data Exchange (ETDEWEB)

    Tong, Wei [Iowa State Univ., Ames, IA (United States)

    1997-10-08

    Microscale separation and detection methods for real-time monitoring of dynamic cellular processes (e.g., secretion) by capillary electrophoresis (CE) and microscopic imaging were developed. Ultraviolet laser-induced native fluorescence (LINF) provides simple, sensitive and direct detection of neurotransmitters and proteins without any derivatization. An on-column CE-LINF protocol for quantification of the release from single cell was demonstrated. Quantitative measurements of both the amount of insulin released from and the amount remaining in the cell (βTC3) were achieved simultaneously. Secretion of catecholamines (norepinephrine (NE) and epinephrine (E)) from individual bovine adrenal chromaffin cells was determined using the on-column CE-LINF. Direct visualization of the secretion process of individual bovine adrenal chromaffin cells was achieved by LINF imaging microscopy with high temporal and spatial resolution. The secretion of serotonin from individual leech Retzius neurons was directly characterized by LINF microscopy with high spatial resolution.

  5. Welcome to Methods and Applications in Fluorescence

    Science.gov (United States)

    Birch, David; Mély, Yves; Wolfbeis, Otto S.

    2013-03-01

    On behalf of the Editorial Board of Methods and Applications in Fluorescence and IOP Publishing we are delighted to invite you to read the first articles in our new journal. Methods and Applications in Fluorescence is forged out of the renowned MAF conference series of the same name and we fully expect the natural synergy between the two to provide the ideal platform for moving the field of fluorescence forward. Our aim is for this new journal to reflect the truly global and diverse impact fluorescence is having across many disciplines and help fluorescence achieve its full potential. Just as MAF is the leading conference in fluorescence we are confident of the high impact of this new journal. Methods and Applications in Fluorescence has a distinguished Editorial Board that is drawn from the MAF conference Permanent Steering Committee. Together with the Editorial Board and the rest of the community, the journal will closely track the very latest developments in fluorescence while delivering a fair and constructive review process. We are very pleased that this journal is backed by the Institute of Physics, one of the world's premier learned societies. IOP Publishing has a wealth of experience in science publishing that dates back to 1874. It is a not-for-profit organization that publishes over 60 journals, many on multidisciplinary topics and many including seminal contributions from Nobel Laureates. Any funding surplus generated by IOP Publishing goes directly back into science through the Institute of Physics, thus helping to nurture science for future generations. We invite submissions as regular articles, review articles and technical notes within the scope of the journal, which includes all the major aspects of fluorescence. This covers both theory and experiment across spectroscopy, imaging, materials, labels, probes and sensors. The applications of fluorescence to emerging areas in bionanotechnology, nanotechnology and medicine are very much part of the

  6. Influence of the gaseous mixture composition on accuracy of molecular iodine on-line detection by laser-induced fluorescence method

    Science.gov (United States)

    Kireev, S. V.; Shnyrev, S. L.

    2016-07-01

    This paper informs on research into the influence of the composition of gaseous mixtures analyzed on the accuracy of on-line molecular iodine detection by laser-induced fluorescence in various gaseous media—in atmospheric air and in technological mixtures formed during reprocessing of spent nuclear fuel. The paper shows that by considering the composition of buffer media and parts of its components, the accuracy of iodine content measurement may be increased in several times.

  7. Remote Sensing and Modelling of Solar Induced Fluorescence

    Science.gov (United States)

    Maier, S. W.

    . For synoptic, remote measurements of the physiological status of green plants under environmental conditions, the fluorescence LIDAR technique was introduced. The first approaches were oriented towards the laser induced fluorescence spectroscopy in order to derive from spectral information parameters related to the physiology of plants. Due to the limited spatial coverage and the complexity of an airborne fluorescence LIDAR these systems were not widely accepted for operational purposes. The objective of this paper is to present a new remote sensing technique that allows to monitor the sun induced chlorophyll fluorescence of green vegetation superimposed to the reflectance spectra. In contrast to the method using the Fraunhofer line in the red spectral region for monitoring the chlorophyll fluorescence the new method uses the spectral absorption feature of atmospheric oxygen at 762nm which better fits to the wavelength of chlorophyll fluorescence than the Fraunhofer absorption line at 656nm. In addition the requirements for spectral resolution and system stability are not so severe for the new method than for the Fraunhofer Lines Discriminator method. It is shown how the sun induced chlorophyll fluorescence can be derived from reflectance spectra measured on ground and from aircraft. The new method can even be applied to medium spectral resolution imaging spectrometers like the airborne sensor ROSIS and AVIRIS or the satellite sensors like MOS, MERIS and CHRIS with a spectral resolution better than 10nm in the oxygen absorption band. Our findings show that sun induced chlorophyll fluorescence can be observed under natural conditions in the NIR spectral region using the absorption band of oxygen on the leaf and the canopy level from ground based and airborne radiance measurements. The results demonstrate that sun induced chlorophyll fluorescence gives a different information than commonly used vegetation indices.

  8. Inositol phosphates induce DAPI fluorescence shift.

    Science.gov (United States)

    Kolozsvari, Bernadett; Parisi, Federica; Saiardi, Adolfo

    2014-06-15

    The polymer inorganic polyP (polyphosphate) and inositol phosphates, such as IP6 (inositol hexakisphosphate; also known as phytic acid), share many biophysical features. These similarities must be attributed to the phosphate groups present in these molecules. Given the ability of polyP to modify the excitation-emission spectra of DAPI we decided to investigate whether inositol phosphates possess the same property. We discovered that DAPI-IP6 complexes emit at approximately 550 nm when excited with light of wavelength 410-420 nm. IP5 (inositol pentakisphosphate) is also able to induce a similar shift in DAPI fluorescence. Conversely, IP3 (inositol trisphosphate) and IP4 (inositol tetrakisphosphate) are unable to shift DAPI fluorescence. We have employed this newly discovered feature of DAPI to study the enzymatic activity of the inositol polyphosphate multikinase and to monitor phytase phosphatase reactions. Finally, we used DAPI-IP6 fluorescence to determine the amount of IP6 in plant seeds. Using an IP6 standard curve this straight-forward analysis revealed that among the samples tested, borlotti beans possess the highest level of IP6 (9.4 mg/g of dry mass), whereas the Indian urad bean the lowest (3.2 mg/g of dry mass). The newly identified fluorescence properties of the DAPI-IP5 and DAPI-IP6 complexes allow the levels and enzymatic conversion of these two important messengers to be rapidly and reliably monitored.

  9. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  10. Laser induced fluorescence of dental caries

    Science.gov (United States)

    Albin, S.; Byvik, C. E.; Buoncristiani, A. M.

    1988-01-01

    Significant differences between the optical spectra taken from sound regions of teeth and carious regions have been observed. These differences appear both in absorption and in laser induced fluorescence spectra. Excitation by the 488 nm line of an argon ion laser beam showed a peak in the emission intensity around 553 nm for the sound dental material while the emission peak from the carious region was red-shifted by approximately 40 nm. The relative absorption of carious region was significantly higher at 488 nm; however its fluorescence intensity peak was lower by an order of magnitude compared to the sound tooth. Implications of these results for a safe, reliable and early detection of dental caries are discussed.

  11. The study of blue LED to induce fluorescence spectroscopy and fluorescence imaging for oral carcinoma detection

    Science.gov (United States)

    Zheng, Longjiang; Hu, Yuanting

    2009-07-01

    Fluorescence spectroscopy and fluorescence imaging diagnosis of malignant lesions provides us with a new method to diagnose diseases in precancerous stage. Early diagnosis of disease has significant importance in cancer treatment, because most cancers can be cured well in precancerous, especially when the diffusion of cancer is limited in a restricted region. In this study, Golden hamster models were applied to 5% 9, 10 dimethyl-1, 2-benzanthracene (DMBA) to induce hamster buccal cheek pouch carcinoma three times a week. Rose Bengal, which has been used in clinican for years and avoids visible side-effect to human was chosen as photosensitizer. 405 nm blue LED was used to induce the fluorescence of photosensitizer. After topical application of photosensitizer, characteristic red emission fluorescence peak was observed around 600nm. Similar, normal oral cavity has special luminescence around 480nm. Fluorescence spectroscopy technology is based on analysing emission peaks of photosensitizer in the areas of oral carcinoma, moreover, red-to-green (IR/IG) intensity ratio is also applied as a diagnostic algorithm. A CCD which is connected with a computer is used to take pictures at carcinoma areas through different filters. Fluorescence images from normal hamster buccal cheek pouch are compared with those from carcinogen-induced models of carcinoma, and morphological differences between normal and lesion tissue can be distinguished. The pictures are analyzed by Matlab and shown on the screen of computer. This paper demonstrates that Rose Bengal could be used as photosensitizer to detect oral carcinoma, and blue LED as excitation source could not only have a good effect to diagnose oral carcinoma, but also decrease cost greatly.

  12. Fluorescent multiplex cell flow systems and methods

    KAUST Repository

    Merzaban, Jasmeen

    2017-06-01

    Systems and methods are provided for simultaneously assaying cell adhesion or cell rolling for multiple cell specimens. One embodiment provides a system for assaying adhesion or cell rolling of multiple cell specimens that includes a confocal imaging system containing a parallel plate flow chamber, a pump in fluid communication with the parallel plate flow chamber via a flow chamber inlet line and a cell suspension in fluid communication with the parallel plate flow chamber via a flow chamber outlet line. The system also includes a laser scanning system in electronic communication with the confocal imaging system, and a computer in communication with the confocal imaging system and laser scanning system. In certain embodiments, the laser scanning system emits multiple electromagnetic wavelengths simultaneously it cause multiple fluorescent labels having different excitation wavelength maximums to fluoresce. The system can simultaneously capture real-time fluorescence images from at least seven cell specimens in the parallel plate flow chamber.

  13. Novel fluorescence molecular imaging of chemotherapy-induced intestinal apoptosis

    Science.gov (United States)

    Levin, Galit; Shirvan, Anat; Grimberg, Hagit; Reshef, Ayelet; Yogev-Falach, Merav; Cohen, Avi; Ziv, Ilan

    2009-09-01

    Chemotherapy-induced enteropathy (CIE) is one of the most serious complications of anticancer therapy, and tools for its early detection and monitoring are highly needed. We report on a novel fluorescence method for detection of CIE, based on molecular imaging of the related apoptotic process. The method comprises systemic intravenous administration of the ApoSense fluorescent biomarker (N,N'-didansyl-L-cystine DDC) in vivo and subsequent fluorescence imaging of the intestinal mucosa. In the reported proof-of-concept studies, mice were treated with either taxol+cyclophosphamide or doxil. DDC was administered in vivo at various time points after drug administration, and tracer uptake by ileum tissue was subsequently evaluated by ex vivo fluorescent microscopy. Chemotherapy caused marked and selective uptake of DDC in ileal epithelial cells, in correlation with other hallmarks of apoptosis (i.e., DNA fragmentation and Annexin-V binding). Induction of DDC uptake occurred early after chemotherapy, and its temporal profile was parallel to that of the apoptotic process, as assessed histologically. DDC may therefore serve as a useful tool for detection of CIE. Future potential integration of this method with fluorescent endoscopic techniques, or development of radio-labeled derivatives of DDC for emission tomography, may advance early diagnosis and monitoring of this severe adverse effect of chemotherapy.

  14. Laser induced fluorescence technique for environmental applications

    Science.gov (United States)

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

    2014-08-01

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

  15. Designing the method for optical in vitro monitoring of the cell-mediated scaffold technology for bone regeneration based on laser-induced fluorescence spectroscopy

    Science.gov (United States)

    Larionov, P. M.; Maslov, N. A.; Papaeva, E. O.; Tereshchenko, V. P.; Khlestkin, V. K.; Bogachev, S. S.; Proskurina, A. S.; Titov, A. T.; Filipenko, M. L.; Pavlov, V. V.; Kudrov, G. A.; Orishich, A. M.

    2016-08-01

    One of the main unsolved problems in traumatology and orthopedics is reconstruction of critical-sized segmental bone defects. We believe that implementation of noninvasive monitoring of the bioengineering stages for cell-mediated bone scaffold by laser-induced fluorescence (LIF) can become a positive aspect in mastering this technique. An electrospun scaffold model (parameters: 10 wt. % polycaprolactone; 5% wt type A gelatin; mean fiber diameter 877.1 ± 169.1, and contact angle 45.3°) seeded with BHK IR cell culture (182 ± 38 cells/mm2) was used to show the principal possibility of differentiating between the scaffold seeded and unseeded with cells. First of all, the fluorescence spectra of the cell-seeded scaffold contain a peak at 305 nm for the excitation range of 230-290 nm, which can be used to differentiate between the samples. An increase in fluorescence intensity of the cell-seeded scaffold in the range of 400- 580 nm upon excitation at 230-340 nm is also noticeable. The wavelength of 250 nm is characterized by high signal intensity and is most suitable for differentiation between the samples.

  16. The Lantibiotic Nisin Induces Transmembrane Movement of a Fluorescent Phospholipid

    NARCIS (Netherlands)

    Moll, Gert N.; Konings, Wil N.; Driessen, Arnold J.M.

    1998-01-01

    Nisin is a pore-forming antimicrobial peptide. The capacity of nisin to induce transmembrane movement of a fluorescent phospholipid in lipid vesicles was investigated. Unilamellar phospholipid vesicles that contained a fluorescent phospholipid

  17. Direct probing of chromatography columns by laser-induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    McGuffin, V.L.

    1992-12-07

    This report summarizes the progress and accomplishments of this research project from September 1, 1989 to February 28, 1993. During this period, we have accomplished all of the primary scientific objectives of the research proposal: (1) constructed and evaluated a laser-induced fluorescence detection system that allows direct examination of the chromatographic column, (2) examined nonequilibrium processes that occur upon solute injection and elution, (3) examined solute retention in liquid chromatography as a function of temperature and pressure, (4) examined solute zone dispersion in liquid chromatography as a function of temperature and pressure, and (5) developed appropriate theoretical models to describe these phenomena. In each of these studies, substantial knowledge has been gained of the fundamental processes that are responsible for chromatographic separations. In addition to these primary research objectives, we have made significant progress in three related areas: (1) examined pyrene as a fluorescent polarity probe insupercritical fluids and liquids as a function of temperature and pressure, (2) developed methods for the class-selective identification of polynuclear aromatic hydrocarbons in coal-derived fluids by microcolumn liquid chromatography with fluorescence quenching detection, and (3) developed methods for the determination of saturated and unsaturated (including omega-3) fatty acids in fish oil extracts by microcolumn liquid chromatography with laser-induced fluorescence detection. In these studies, the advanced separation and detection techniques developed in our laboratory are applied to practical problems of environmental and biomedical significance.

  18. Direct probing of chromatography columns by laser-induced fluorescence

    Science.gov (United States)

    McGuffin, V. L.

    1992-12-01

    This report summarizes the progress and accomplishments of this research project from 1 Sep. 1989 to 28 Feb. 1993. During this period, we have accomplished all of the primary scientific objectives of the research proposal: (1) constructed and evaluated a laser-induced fluorescence detection system that allows direct examination of the chromatographic column, (2) examined nonequilibrium processes that occur upon solute injection and elution, (3) examined solute retention in liquid chromatography as a function of temperature and pressure, (4) examined solute zone dispersion in liquid chromatography as a function of temperature and pressure, and (5) developed appropriate theoretical models to describe these phenomena. In each of these studies, substantial knowledge has been gained of the fundamental processes that are responsible for chromatographic separations. In addition to these primary research objectives, we have made significant progress in three related areas: (1) examined pyrene as a fluorescent polarity probe in supercritical fluids and liquids as a function of temperature and pressure, (2) developed methods for the class-selective identification of polynuclear aromatic hydrocarbons in coal-derived fluids by microcolumn liquid chromatography with fluorescence quenching detection, and (3) developed methods for the determination of saturated and unsaturated (including omega-3) fatty acids in fish oil extracts by microcolumn liquid chromatography with laser-induced fluorescence detection. In these studies, the advanced separation and detection techniques developed in our laboratory are applied to practical problems of environmental and biomedical significance.

  19. Characterisation of estuarine intertidal macroalgae by laser-induced fluorescence

    Science.gov (United States)

    Gameiro, Carla; Utkin, Andrei B.; Cartaxana, Paulo

    2015-12-01

    The article reports the application of laser-induced fluorescence (LIF) for the assessment of macroalgae communities of estuarine intertidal areas. The method was applied for the characterisation of fifteen intertidal macroalgae species of the Tagus estuary, Portugal, and adjacent coastal area. Three bands characterised the LIF spectra of red macroalgae with emission maxima in the ranges 577-583 nm, 621-642 nm and 705-731 nm. Green and brown macroalgae showed one emission maximum in the red region (687-690 nm) and/or one in the far-red region (726-732 nm). Characteristics of LIF emission spectra were determined by differences in the main fluorescing pigments: phycoerythrin, phycocyanin and chlorophyll a (Chl a). In the green and brown macroalgae groups, the relative significance of the two emission maxima seems to be related to the thickness of the photosynthetic layer. In thick macroalgae, like Codium tomentosum or Fucus vesiculosus, the contribution of the far-red emission fluorescence peak was more significant, most probably due to re-absorption of the emitted red Chl a fluorescence within the dense photosynthetic layer. Similarly, an increase in the number of layers of the thin-blade green macroalgae Ulva rigida caused a shift to longer wavelengths of the red emission maximum and the development of a fluorescence peak at the far-red region. Water loss from Ulva's algal tissue also led to a decrease in the red/far-red Chl fluorescence ratio (F685/F735), indicating an increase in the density of chloroplasts in the shrinking macroalgal tissue during low tide exposure.

  20. Characterization of Cr(V)-induced genotoxicity using CdTe nanocrystals as fluorescent probes.

    Science.gov (United States)

    Zhang, Wen-Hao; Sui, Chao-Xia; Wang, Xie; Yin, Gong-Ju; Liu, Ying-Fan; Zhang, Ding

    2014-12-21

    CdTe nanocrystals capped by cysteamine were synthesized to study Cr(V)-induced genotoxicity. On the surface of TiO2 thin films, the stepwise process of DNA breakage induced by Cr(V)-GSH complexes was vividly observed by using CdTe-DNA self-assembled fluorescent probes; in acetate buffer solution, an analytical method was developed to detect Cr(V)-induced genotoxicity with CdTe fluorescent probes.

  1. Ultrafast laser-collision-induced fluorescence in atmospheric pressure plasma

    Science.gov (United States)

    Barnat, E. V.; Fierro, A.

    2017-04-01

    The implementation and demonstration of laser-collision-induced fluorescence (LCIF) generated in atmospheric pressure helium environments is presented in this communication. As collision times are observed to be fast (~10 ns), ultrashort pulse laser excitation (<100 fs) of the 23S to 33P (388.9 nm) is utilized to initiate the LCIF process. Both neutral-induced and electron-induced components of the LCIF are observed in the helium afterglow plasma as the reduced electric field (E/N) is tuned from  <0.1 Td to over 5 Td. Under the discharge conditions presented in this study (640 Torr He), the lower limit of electron density detection is ~1012 e cm‑3. The spatial profiles of the 23S helium metastable and electrons are presented as functions of E/N to demonstrate the spatial resolving capabilities of the LCIF method.

  2. Proton-induced x-ray fluorescence CT imaging

    Energy Technology Data Exchange (ETDEWEB)

    Bazalova-Carter, Magdalena, E-mail: bazalova@stanford.edu; Xing, Lei [Department of Radiation Oncology, Stanford University, Stanford, California 94305-5847 and Global Station for Quantum Medical Science and Engineering, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo 060-8648 (Japan); Ahmad, Moiz [Department of Radiation Oncology, Stanford University, Stanford, California 94305-5847 (United States); Matsuura, Taeko; Takao, Seishin; Shirato, Hiroki; Umegaki, Kikuo [Department of Medical Physics, Proton Beam Therapy Center, Hokkaido University Hospital, Sapporo 060-8648, Japan and Global Station for Quantum Medical Science and Engineering, Global Institution for Collaborative Research and Education (GI-CoRE), Hokkaido University, Sapporo 060-8648 (Japan); Matsuo, Yuto [Department of Medical Physics, Proton Beam Therapy Center, Hokkaido University Hospital, Sapporo 060-8648 (Japan); Fahrig, Rebecca [Department of Radiology, Stanford University, Stanford, California 94305 (United States)

    2015-02-15

    Purpose: To demonstrate the feasibility of proton-induced x-ray fluorescence CT (pXFCT) imaging of gold in a small animal sized object by means of experiments and Monte Carlo (MC) simulations. Methods: First, proton-induced gold x-ray fluorescence (pXRF) was measured as a function of gold concentration. Vials of 2.2 cm in diameter filled with 0%–5% Au solutions were irradiated with a 220 MeV proton beam and x-ray fluorescence induced by the interaction of protons, and Au was detected with a 3 × 3 mm{sup 2} CdTe detector placed at 90° with respect to the incident proton beam at a distance of 45 cm from the vials. Second, a 7-cm diameter water phantom containing three 2.2-diameter vials with 3%–5% Au solutions was imaged with a 7-mm FWHM 220 MeV proton beam in a first generation CT scanning geometry. X-rays scattered perpendicular to the incident proton beam were acquired with the CdTe detector placed at 45 cm from the phantom positioned on a translation/rotation stage. Twenty one translational steps spaced by 3 mm at each of 36 projection angles spaced by 10° were acquired, and pXFCT images of the phantom were reconstructed with filtered back projection. A simplified geometry of the experimental data acquisition setup was modeled with the MC TOPAS code, and simulation results were compared to the experimental data. Results: A linear relationship between gold pXRF and gold concentration was observed in both experimental and MC simulation data (R{sup 2} > 0.99). All Au vials were apparent in the experimental and simulated pXFCT images. Specifically, the 3% Au vial was detectable in the experimental [contrast-to-noise ratio (CNR) = 5.8] and simulated (CNR = 11.5) pXFCT image. Due to fluorescence x-ray attenuation in the higher concentration vials, the 4% and 5% Au contrast were underestimated by 10% and 15%, respectively, in both the experimental and simulated pXFCT images. Conclusions: Proton-induced x-ray fluorescence CT imaging of 3%–5% gold solutions in a

  3. Binding-Induced Fluorescence of Serotonin Transporter Ligands

    DEFF Research Database (Denmark)

    Wilson, James; Ladefoged, Lucy Kate; Babinchak, Michael

    2014-01-01

    The binding-induced fluorescence of 4-(4-(dimethylamino)-phenyl)-1-methylpyridinium (APP(+)) and two new serotonin transporter (SERT)-binding fluorescent analogues, 1-butyl-4-[4-(1-dimethylamino)phenyl]-pyridinium bromide (BPP(+)) and 1-methyl-4-[4-(1-piperidinyl)phenyl]-pyridinium (PPP(+)), has...

  4. ALA-induced PpIX fluorescence in epileptogenic tissue

    Science.gov (United States)

    Kleen, Jonathan K.; Valdes, Pablo A.; Harris, Brent T.; Holmes, Gregory L.; Paulsen, Keith D.; Roberts, David W.

    2011-03-01

    Astrogliotic tissue displays markedly increased levels of ALA-induced PpIX fluorescence, making it useful for fluorescence-guided resection in glioma surgery. In patients with temporal lobe epilepsy (TLE) and corresponding animal models, there are areas of astrogliosis that often co-localize with the epileptic focus, which can be resected to eliminate seizures in the majority of treated patients. If this epileptogenic tissue can exhibit PpIX fluorescence that is sufficiently localized, it could potentially help identify margins in epilepsy surgery. We tested the hypothesis that ALA-induced PpIX fluorescence could visually accentuate epileptogenic tissue, using an established animal model of chronic TLE. An acute dose of pilocarpine was used to induce chronic seizure activity in a rat. This rat and a normal control were given ALA, euthanized, and brains examined post-mortem for PpIX fluorescence and neuropathology. Preliminary evidence indicates increased PpIX fluorescence in areas associated with chronic epileptic changes and seizure generation in TLE, including the hippocampus and parahippocampal areas. In addition, strong PpIX fluorescence was clearly observed in layer II of the piriform cortex, a region known for epileptic reorganization and involvement in the generation of seizures in animal studies. We are further investigating whether ALA-induced PpIX fluorescence can consistently identify epileptogenic zones, which could warrant the extension of this technique to clinical studies for use as an adjuvant guidance technology in the resection of epileptic tissue.

  5. The Lantibiotic Nisin Induces Transmembrane Movement of a Fluorescent Phospholipid

    NARCIS (Netherlands)

    Moll, Gert N.; Konings, Wil N.; Driessen, Arnold J.M.

    1998-01-01

    Nisin is a pore-forming antimicrobial peptide. The capacity of nisin to induce transmembrane movement of a fluorescent phospholipid in lipid vesicles was investigated. Unilamellar phospholipid vesicles that contained a fluorescent phospholipid (1-acyl-2-{6-[(7-nitro-2-1,3-benzoxadiazol-4-yl)amino]ca

  6. Fluorescent method for monitoring cheese starter permeabilization and lysis

    NARCIS (Netherlands)

    Bunthof, C.J.; Schalkwijk, van S.; Meijer, W.; Abee, T.; Hugenholtz, J.

    2001-01-01

    A fluorescence method to monitor lysis of cheese starter bacteria using dual staining with the LIVE/DEAD BacLight bacterial viability kit is described. This kit combines membrane-permeant green fluorescent nucleic acid dye SYTO 9 and membrane-impermeant red fluorescent nucleic acid dye propidium iod

  7. Diagnosis of dental caries using quantitative light-induced fluorescence

    Science.gov (United States)

    Amaechi, Bennett T.; Higham, Susan M.

    2001-10-01

    Current dental diagnostic methods can detect caries but cannot quantify the mineral status of the lesion. Quantitative Light-induced Fluorescence (QLF) measures the percentage fluorescence radiance change of demineralised enamel with respect to surround sound enamel, and related it directly to the amount of mineral lost during demineralisation. Demineralisation of teeth to produce caries-like lesions and the subsequent remineralisation of the lesions were monitored quantitatively and longitudinally with QLF. The influence of factors such as presence of plaque or saliva, lesion staining, lesion magnification, tooth thickness and developmental hypomineralisation, on the reproducibility of QLF imaging and analysis were investigated, Results showed that the integrated fluorescence change (hence the mineral loss) increased linearly with demineralisation time and decreased with increasing remineralisation time. Caries detection was limited by saliva or plaque, but enhanced by staining. QLF could not discriminate between developmental hypomineralisation and caries. Neither the variation in tooth thickness nor lesion magnification within the limit of a sharp image made a significant difference in QLF analysis. It was concluded that QLF could detect and quantitatively monitor the mineral changes in an incipient caries on a longitudinal basis, however detection may be limited by the presence of saliva or plaque or enhanced by staining.

  8. Evanescent wave induced fluorescence. A tool for quantitative interfacial analysis

    CERN Document Server

    Byrne, C D

    2000-01-01

    Time-resolved angle-resolved evanescent wave induced fluorescence spectroscopy (EWIFS) has been used, for the first time, to determine interfacial concentration distributions of molecular species. Theoretical calculations demonstrate that in dynamic systems the non-radiative fluorescence decay coefficients of molecular species are effected only in a minor way by the presence of a dielectric interface. Consequently, measurements of interfacial fluorescence decay times are used to probe variations in molecular fluorescence quantum efficiencies, caused by the presence of an interface. The understanding of these variations is combined with angle-resolved evanescent wave theory. Examination of derived theoretical models using simulated data demonstrates that angle-resolved EWIFS is capable of measuring interfacial interactions on a nanometer scale. An evanescent wave induced fluorescence spectrometer is designed and fabricated to allow the measurement of the time-integrated and time-resolved interfacial emission. ...

  9. Induced systemic resistance by fluorescent Pseudomonas spp.

    NARCIS (Netherlands)

    Bakker, P.A.H.M.; Pieterse, C.M.J.; Loon, L.C. van

    2007-01-01

    Fluorescent Pseudomonas spp. have been studied for decades for their plant growth-promoting effects through effective suppression of soilborne plant diseases. The modes of action that play a role in disease suppression by these bacteria include siderophore-mediated competition for iron, antibiosis,

  10. Estimating Chlorophyll Fluorescence Parameters Using the Joint Fraunhofer Line Depth and Laser-Induced Saturation Pulse (FLD-LISP Method in Different Plant Species

    Directory of Open Access Journals (Sweden)

    Parinaz Rahimzadeh-Bajgiran

    2017-06-01

    Full Text Available A comprehensive evaluation of the recently developed Fraunhofer line depth (FLD and laser-induced saturation pulse (FLD-LISP method was conducted to measure chlorophyll fluorescence (ChlF parameters of the quantum yield of photosystem II (ΦPSII, non-photochemical quenching (NPQ, and the photosystem II-based electron transport rate (ETR in three plant species including paprika (C3 plant, maize (C4 plant, and pachira (C3 plant. First, the relationships between photosynthetic photon flux density (PPFD and ChlF parameters retrieved using FLD-LISP and the pulse amplitude-modulated (PAM methods were analyzed for all three species. Then the relationships between ChlF parameters measured using FLD-LISP and PAM were evaluated for the plants in different growth stages of leaves from mature to aging conditions. The relationships of ChlF parameters/PPFD were similar in both FLD-LISP and PAM methods in all plant species. ΦPSII showed a linear relationship with PPFD in all three species whereas NPQ was found to be linearly related to PPFD in paprika and maize, but not for pachira. The ETR/PPFD relationship was nonlinear with increasing values observed for PPFDs lower than about 800 μmol m−2 s−1 for paprika, lower than about 1200 μmol m−2 s−1 for maize, and lower than about 800 μmol m−2 s−1 for pachira. The ΦPSII, NPQ, and ETR of both the FLD-LISP and PAM methods were very well correlated (R2 = 0.89, RMSE = 0.05, (R2 = 0.86, RMSE = 0.44, and (R2 = 0.88, RMSE = 24.69, respectively, for all plants. Therefore, the FLD-LISP method can be recommended as a robust technique for the estimation of ChlF parameters.

  11. Quenching of chlorophyll fluorescence induced by silver nanoparticles

    Science.gov (United States)

    Queiroz, A. M.; Mezacasa, A. V.; Graciano, D. E.; Falco, W. F.; M'Peko, J.-C.; Guimarães, F. E. G.; Lawson, T.; Colbeck, I.; Oliveira, S. L.; Caires, A. R. L.

    2016-11-01

    The interaction between chlorophyll (Chl) and silver nanoparticles (AgNPs) was evaluated by analyzing the optical behavior of Chl molecules surrounded by different concentrations of AgNPs (10, 60, and 100 nm of diameter). UV-Vis absorption, steady state and time-resolved fluorescence measurements were performed for Chl in the presence and absence of these nanoparticles. AgNPs strongly suppressed the Chl fluorescence intensity at 678 nm. The Stern-Volmer constant (KSV) showed that fluorescence suppression is driven by the dynamic quenching process. In particular, KSV was nanoparticle size-dependent with an exponential decrease as a function of the nanoparticle diameter. Finally, changes in the Chl fluorescence lifetime in the presence of nanoparticles demonstrated that the fluorescence quenching may be induced by the excited electron transfer from the Chl molecules to the metal nanoparticles.

  12. Laser-induced fluorescence detection of stomach cancer using hypericin

    Science.gov (United States)

    Dets, Sergiy M.; Buryi, Alexander N.; Melnik, Ivan S.; Joffe, Alexander Y.; Rusina, Tatyana V.

    1996-12-01

    Natural photodynamic pigment hypericin having intrinsic antitumor properties was applied for fluorescence detection of cancer. Clinical investigation of hypericin was performed to ensure high tumor/normal fluorescence contrast in digestion organs. Laser-induced autofluorescence and exogenous fluorescence analysis of normal tissue and stomach adenocarcinoma was performed using helium-cadmium laser (8 mW, 442 nm). Twenty-one patients have undergone procedure of fluorescence detection of tumors before and after photosensitization. For sensitization of patients we used five or seven capsules containing hypericin in amount of 1 mg which have been administered orally. Strong yellow-red fluorescence of hypericin in tissue with maximum at 603 nm and autofluorescence peak at 535 nm gives an intensity ratio I(603 nm)/I(535 nm) of 2 - 2.5 from cancerous tissue and provides 85% specificity. Preliminary in vivo results of auto- and fluorescence analysis using hypericin photosensitization from one patient with esophageal cancer and eleven patients with stomach cancer proven histologically are encouraging and indicate the high reliability of laser-induced fluorescence technique with hypericin in detection of early stage malignant lesions.

  13. Analyzing of LED-induced Blood Fluorescent Spectra

    Institute of Scientific and Technical Information of China (English)

    高淑梅; 骆晓森; 兰秀风; 刘莹; 陆健; 倪晓武

    2002-01-01

    The visible fluorescence spectra of healthy mice blood cells have been measured by light emitting diode (LED) excitation at about 570 nm (Δλ1/2≈32 nm ) in vitro. It is found that the spectral profiles of mouse blood, erythrocyte and hemoglobin are similar. The physical mechanism for LED-induced blood cells fluorescence spectra is analyzed. The development of low intensity laser or light irradiating blood therapy in vivo and in vitro may be guided by the understanding of blood fluorescence characteristics.

  14. Laser induced fluorescence photobleaching anemometer for microfluidic devices.

    Science.gov (United States)

    Wang, G R

    2005-04-01

    We have developed a novel, non-intrusive fluid velocity measurement method based on photobleaching of a fluorescent dye for microfluidic devices. The residence time of the fluorescent dye in a laser beam depends on the flow velocity and approximately corresponds to the decaying time of the photobleaching of the dye in the laser beam. The residence time is inversely proportional to the flow velocity. The fluorescence intensity increases with the flow velocity due to the decrease of the residence time. A calibration curve between fluorescence intensity and known flow velocity should be obtained first. The calibration relationship is then used to calculate the flow velocity directly from the measured fluorescence intensity signal. The new method can measure the velocity very quickly and is easy to use. It is demonstrated for both pressure driven flow and electroosmotic flow.

  15. Evaluation of immunoglobulins in bovine colostrum using laser induced fluorescence.

    Science.gov (United States)

    Abdel-Salam, Z; Abdel Ghany, Sh; Harith, M A

    2014-11-01

    The objective of the present study was to exploit laser induced fluorescence (LIF) as a spectrochemical analytical technique for evaluation of immunoglobulin (IgG) in bovine colostrum. Colostrum samples were collected from different American Holstein cows at different times after calving. Four samples were gathered from each cow; the first three samples were obtained from the first three milkings (colostrum) and the fourth sample (milk) was obtained a week after calving. It has been demonstrated that LIF can be used as a simple, fast, sensitive and less costly spectrochemical analytical technique for qualitative estimation of IgG in colostrum. LIF results have been confirmed via the quantitative evaluation of IgG in the same samples adopting the single radial immunodiffusion conventional technique and a very good agreement has been obtained. Through LIF it was possible to evaluate bovine colostrum after different milking times and to differentiate qualitatively between colostrum from different animals which may reflect their general health status. A fluorescence linear calibration curve for IgG concentrations from 0 up to 120 g L(-1) has been obtained. In addition, it is feasible to adopt this technique for in situ measurements, i.e. in dairy cattle farms as a simple and fast method for evaluation of IgG in bovine colostrum instead of using lengthy and complicated conventional techniques in laboratories.

  16. Stink Bug Feeding Induces Fluorescence in Developing Cotton Bolls

    Directory of Open Access Journals (Sweden)

    Toews Michael D

    2011-08-01

    Full Text Available Abstract Background Stink bugs (Hemiptera: Pentatomidae comprise a critically important insect pest complex affecting 12 major crops worldwide including cotton. In the US, stink bug damage to developing cotton bolls causes boll abscission, lint staining, reduced fiber quality, and reduced yields with estimated losses ranging from 10 to 60 million dollars annually. Unfortunately, scouting for stink bug damage in the field is laborious and excessively time consuming. To improve scouting accuracy and efficiency, we investigated fluorescence changes in cotton boll tissues as a result of stink bug feeding. Results Fluorescent imaging under long-wave ultraviolet light showed that stink bug-damaged lint, the inner carpal wall, and the outside of the boll emitted strong blue-green fluorescence in a circular region near the puncture wound, whereas undamaged tissue emissions occurred at different wavelengths; the much weaker emission of undamaged tissue was dominated by chlorophyll fluorescence. We further characterized the optimum emission and excitation spectra to distinguish between stink bug damaged bolls from undamaged bolls. Conclusions The observed characteristic fluorescence peaks associated with stink bug damage give rise to a fluorescence-based method to rapidly distinguish between undamaged and stink bug damaged cotton bolls. Based on the fluorescent fingerprint, we envision a fluorescence reflectance imaging or a fluorescence ratiometric device to assist pest management professionals with rapidly determining the extent of stink bug damage in a cotton field.

  17. Laser induced fluorescence applied to studies of particle behaviour in high-temperature plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Muraoka, K.; Uchino, K.; Kajiwara, T.; Maeda, M.; Okada, T. [Kyushu Univ., Fukuoka (Japan)

    1995-03-01

    In this paper, we first review the principle of Laser Induced Fluorescence (LIF), then give an overview of tunable laser sources, the crucial hardware for the experiment, and describe methods of calibration to obtain necessary information from the observed fluorescence, followed by the plasma measurements which have already been conducted. Comments are made for the future perspective of LIF for high-temperature plasma diagnostics. (J.P.N.).

  18. An optical method for reducing green fluorescence from urine during fluorescence-guided cystoscopy

    DEFF Research Database (Denmark)

    Lindvold, Lars René; Hermann, Gregers G

    2016-01-01

    Photodynamic diagnosis (PDD) of bladder tumour tissue significantly improves endoscopic diagnosis and treatment of bladder cancer in rigid cystoscopes in the operating theatre and thus reduces tumour recurrence. PDD comprises the use of blue light, which unfortunately excites green fluorescence...... from urine. As this green fluorescence confounds the desired red fluorescence of the PDD, methods for avoiding this situation particularly in cystoscopy using flexible cystoscopes are desirable. In this paper we demonstrate how a tailor made high power LED light source at 525 nm can be used...... for fluorescence assisted tumour detection using both a flexible and rigid cystoscope used in the outpatient department (OPD) and operating room (OR) respectively. It is demonstrated both in vitro and in vivo how this light source can significantly reduce the green fluorescence problem with urine. At the same time...

  19. Identification of cholesterol gallstones using in vitro low-fluence laser-induced fluorescence spectroscopic analysis

    Science.gov (United States)

    Wrobel, R.; Niay, P.; Bernage, P.; Blondeau, J. M.; Ledee, J. J.; Brunetaud, J. M.

    1990-12-01

    Identifying the chemical composition of gallstones may be important in certain cases of calculus biliary disease when planning a dissolution therapy or a fragmentation of the calculi using pulsed lasers. The present study was conducted in vitro to evaluate the feasibility of distinguishing cholesterol gallstones from pigment stones. We propose an identification method in which the stone fluorescence spectrum, induced by a low fluence laser, is recorded using an optical multichannel analyser. Fluorescence spectra of twenty-two stones were recorded together with the fluorescence spectra of various pure compounds likely to compose the gallstones, using successively four different pump lasers (λp=308 nm, 337 nm, 423 nm, 469 nm). The fluorescence spectra of cholesterol gallstones are quite different from the pigment ones. Ratios of fluorescence intensities taken at three different wavelengths enable one to distinguish easily between cholesterol and pigment stones.

  20. Laser-induced fluorescence in diagnosis of dental caries

    Science.gov (United States)

    Drakaki, Eleni A.; Makropoulou, Mersini I.; Khabbaz, Maruan; Serafetinides, Alexandros A.

    2003-09-01

    The autofluorescence spectra of hard dental tissues, both in normal and pathological areas were investigated in this study. The measurements were performed both on the intact hard tissues of the examined teeth, such as enamel, dentine, cementum, and root canal, and on the tissues pathologically affected by caries (superficial, intermediate, and deep). Various laser wavelengths (337 nm, 488 nm, and 514 nm) were used to irradiate the dental surfaces and a computer-controlled spectrograph captured the fluorescent spectra. The emission signals were stored, measured, analyzed and quantified in terms of wavelength distribution and the relative photon intensity. Results indicated that the fluorescent spectra from healthy enamel, dentine, and cementum were almost identical in form, depending on the excitation wavelength. The intact and affected hard tissues were greatly different in the integral fluorescent intensity. Healthy areas were found to produce the most pronounced fluorescent intensity, whereas the carious regions produced the weaker fluorescent intensity. Independently of the laser excitation wavelength, dentin regions were found to produce the most pronounced fluorescent intensity than any other dental component. The fluorescence signal of carious affected dental structure revealed a reed shifted spectral curve, more pronounced after 488 nm excitation. There was a pronounced red shift for deep caries (crown -- root caries), after ultraviolet laser excitation. Excitation with visible wavelengths did not produce such differences between intact and cervical, deep carious affected tissue. Using a monochromatic light source without any light output at the wavelengths of fluorescence, e.g. a laser with the appropriate filters, the difference in fluorescence between intact and carious enamel was generally easy to observe. Finally, we found that the blue line of an argon ion laser is preferable for superficial caries detection, while the ultraviolet emitting nitrogen

  1. Laser-induced fluorescence of oral mucosa cancer

    Science.gov (United States)

    Jaliashvili, Z. V.; Medoidze, T. D.; Melikishvili, Z. G.; Gogilashvili, K. T.

    2017-10-01

    The laser-induced fluorescence (LIF) spectra have been measured for cancer-infused and control mice mucosa tissues. It was established that there is quite a difference between their LIF spectral shapes. These spectral shapes are used to express the diagnostic of different states of tissues: from normal to cancer.

  2. 毛细管电泳分离-激光诱导荧光检测巴氯芬和加巴喷丁%Determination of baclofen and gabapentin by capillary electrophoresis-laser-induced fluorescence method

    Institute of Scientific and Technical Information of China (English)

    曹丽伟; 李聪; 任东

    2011-01-01

    采用新合成的6-氧-(N-琥珀酰亚胺乙酸酯)-9-(2’-甲氧羰基)荧光素(SAMF)作为柱前衍生试剂,用甘氨酸做内标,毛细管电泳分离-激光诱导荧光检测血清中的巴氯芬(baclofen)和加巴喷丁(gabapentin).研究表明,在磷酸盐缓冲溶液(pH=8)介质中,25℃下15 min即可完成衍生反应.以pH =5的30 mmol/L的磷酸盐缓冲溶液作为电泳介质,衍生产物在16 min内达到电泳基线分离,检出限为4×10-10 mol/L.将新建立的方法用于血清中巴氯芬、加巴喷丁的分析测定,回收率为95.5% ~ 102.01%,结果令人满意.%A sensitive and efficient analysis method for determination of baclofen and gabapentin in human serum based on capillary electrophoresis with laser-induced fluorescence (LIF) has been established. 6-Oxy-( N-succinimidylacetate)- 9-(2'-Meth-oxycarbonyl) fluorescein (SAMF), a new synthesized fluorescence reagent, was used for pre-column derivatization of the "non-fluorescent drug in serum. Glycin was used as an internal standard. The best derivation condition was obtained in phosphate buffer (pH =8) at 25℃ for 15 min. Optimal separation and detection were obtained with an background electrolyte of 30 mmol/L phosphate buffer ( pH = 5 ) and LJF was excited at 473 nm. The detection limit of baclofen was 4 x 10-10 mol/L. This sensitive method was effectively used to determine baclofen and gabapentin in human serum with recoveries ranging from 95.5% to 102.01%.

  3. Fluorescence spectroscopic detection of early injury-induced atherosclerosis

    Science.gov (United States)

    Lucas, Alexandra; Perk, Masis; Wen, Yue; Smith, Carol

    1992-08-01

    Laser-induced fluorescence spectroscopy has been used for the detection of advanced atherosclerotic lesions. Angioplasty balloon-mediated injury was examined spectroscopically in order to assess the sensitivity of fluorescence spectroscopy for detection of early atherosclerosis. Abdominal aortic balloon angioplasty was performed via femoral artery cutdown in nine White Leghorn roosters (five normal, four atherogenic diet). Roosters were sacrificed at 1, 2, 4, 8, and 12 week intervals. Fluorescence emission spectra (n equals 114) were recorded from each aortic section (XeCl excimer laser, 308 nm, 1.5 - 2.0 mJ/pulse, 5 Hz). Changes in normalized fluorescence emission intensity were correlated with selected sections of histology. All balloon-injured segments showed intimal fibrous proliferation. For intimal thickness measuring > 70 (mu) , fluorescence emission intensity was decreased at 440 - 460 nm (p Lesions complicated by thrombus also had lower fluorescence emission at 425 - 450 nm when compared to histologically normal aorta (p muscular (abdominal) aorta (p muscular abdominal aorta.

  4. In vivo cellular imaging using fluorescent proteins - Methods and Protocols

    Directory of Open Access Journals (Sweden)

    M. Monti

    2012-12-01

    Full Text Available The discovery and genetic engineering of fluorescent proteins has revolutionized cell biology. What was previously invisible to the cell often can be made visible with the use of fluorescent proteins. With this words, Robert M. Hoffman introduces In vivo Cellular Imaging Using Fluorescent proteins, the eighteen chapters book dedicated to the description of how fluorescence proteins have changed the way to analyze cellular processes in vivo. Modern researches aim to study new and less invasive methods able to follow the behavior of different cell types in different biological contexts: for example, how cancer cells migrate or how they respond to different therapies. Also, in vivo systems can help researchers to better understand animal embryonic development so as how fluorescence proteins may be used to monitor different processes in living organisms at the molecular and cellular level.

  5. The effects of visual fluorescence marking induced by 5-aminolevulinic acid for endoscopic diagnosis of urinary bladder cancer

    Science.gov (United States)

    Daniltchenko, Dmitri I.; Koenig, Frank; Schnorr, Dietmar; Valdman, Alexander; Al-Shukri, Salman; Loening, Stefan A.

    2003-10-01

    During cystoscopy procedure, fluorescence diagnostics induced by 5-ALA improves visual detection of the bladder cancer. Macroscopic ALA-fluorescence allows visualizing of small flat tumors, carcinoma in situ, true neoplasm margins and dysplasias of the bladder. Following ALA instillation, cystoscopy has been performed under both standard and blue light illumination. Totally, 153 biopsies have been carried out at 53 patients with suspicion of bladder cancer. The results were compared to ALA-fluorescence data. In 13% of the patients, bladder cancer and dysplasia were found out in addition, due to red fluorescence. The sensitivity and specificity of ALA-fluorescence technique aggregated 96% and 52% respectively. The sensitivity and specificity of 5-ALA-fluorescent detection exceeded standard endoscopy under white light on 20%. The new method does not exclude a false positive and a false negative fluorescent luminescence. The ALA-based fluorescence detection system enhances the diagnosis of malignant/dysplastic bladder lesions significantly.

  6. In-vivo optical detection of cancer using chlorin e6 – polyvinylpyrrolidone induced fluorescence imaging and spectroscopy

    Directory of Open Access Journals (Sweden)

    Soo Khee

    2009-01-01

    Full Text Available Abstract Background Photosensitizer based fluorescence imaging and spectroscopy is fast becoming a promising approach for cancer detection. The purpose of this study was to examine the use of the photosensitizer chlorin e6 (Ce6 formulated in polyvinylpyrrolidone (PVP as a potential exogenous fluorophore for fluorescence imaging and spectroscopic detection of human cancer tissue xenografted in preclinical models as well as in a patient. Methods Fluorescence imaging was performed on MGH human bladder tumor xenografted on both the chick chorioallantoic membrane (CAM and the murine model using a fluorescence endoscopy imaging system. In addition, fiber optic based fluorescence spectroscopy was performed on tumors and various normal organs in the same mice to validate the macroscopic images. In one patient, fluorescence imaging was performed on angiosarcoma lesions and normal skin in conjunction with fluorescence spectroscopy to validate Ce6-PVP induced fluorescence visual assessment of the lesions. Results Margins of tumor xenografts in the CAM model were clearly outlined under fluorescence imaging. Ce6-PVP-induced fluorescence imaging yielded a specificity of 83% on the CAM model. In mice, fluorescence intensity of Ce6-PVP was higher in bladder tumor compared to adjacent muscle and normal bladder. Clinical results confirmed that fluorescence imaging clearly captured the fluorescence of Ce6-PVP in angiosarcoma lesions and good correlation was found between fluorescence imaging and spectral measurement in the patient. Conclusion Combination of Ce6-PVP induced fluorescence imaging and spectroscopy could allow for optical detection and discrimination between cancer and the surrounding normal tissues. Ce6-PVP seems to be a promising fluorophore for fluorescence diagnosis of cancer.

  7. Capillary electrophoresis with laser-induced fluorescence: environmental applications.

    Science.gov (United States)

    Riddick, Lee; Brumley, William C

    2008-01-01

    Capillary electrophoresis (CE), especially free-zone CE, offers a relatively simple separation with moderate selectivity based on the mobility of ions in solution. Laser-induced fluorescence (LIF) detection, an extremely sensitive technique, can be coupled with a variety of separation conditions to achieve sensitive and quantitative results. When these techniques are combined, CE/LIF provides the sensitivity and increased selectivity that makes trace level environmental analysis of fluorescent compounds possible at or below levels typical for gas chromatography (GC)/mass spectrometry (MS). We offer a panoramic review of the role of these tools in solving environmental and related analytical problems before providing a detailed experimental protocol.

  8. Determination of flame temperature using laser induced fluorescence (LIF)

    Energy Technology Data Exchange (ETDEWEB)

    Silva, Lina Augusta Martins Ramos da [Instituto Tecnologico de Aeronautica (ITA/DCTA), Sao Jose dos Campos, SP (Brazil); Barreta, Gilberto; Carinhana Junior, Dermeval; Toledo, Antonio Osny de [Instituto de Estudos Avancados (IEAv/DCTA), Sao Jose dos Campos, SP (Brazil)

    2011-07-01

    Full text: Laser Induced Fluorescence, LIF, is one of the most used techniques in combustion diagnostics. The excitation method is based on the spontaneous spectrum from atoms or molecules that were excited by laser radiation. In general, intermediated combustion species, as OH and CH radicals, are used as temperature probes. Usually, several rotational levels are used by tuning the laser afterwards across the corresponding absorption transitions. The emission spectrum is detected in a convenient spectral range. The accuracy of measurements depends on the laser linewidth, the delay between the laser excitation and spectra detection and, mainly, the spectral interval of laser excitation. In this work, OH LIF spectra from stoichiometric LPG (Liquefied Petroleum Gas) flames were obtained using a tuned laser with a spectral range set from 282 nm up to 283 nm. This corresponds to the rotation levels of the 0-1 vibration band of OH radical. The laser energy was ca. 0.2 mJ. The emission spectrum was collected in the spectral range of the 305-310 nm, which corresponds to the region of the 0-0 band. The temperature was determined by the Boltzmann method. This is based on the measurement of the relative peak intensities of the emission spectrum. A plot of natural logarithm of line intensities versus energy level returns a straight line, whose slope is the inverse of the rotational temperature. The flame temperature was ca. 2300 K. This value is consistent with previous results obtained in our laboratory. (author)

  9. System and method for measuring fluorescence of a sample

    Energy Technology Data Exchange (ETDEWEB)

    Riot, Vincent J.

    2017-06-27

    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  10. System and method for measuring fluorescence of a sample

    Energy Technology Data Exchange (ETDEWEB)

    Riot, Vincent J

    2015-03-24

    The present disclosure provides a system and a method for measuring fluorescence of a sample. The sample may be a polymerase-chain-reaction (PCR) array, a loop-mediated-isothermal amplification array, etc. LEDs are used to excite the sample, and a photodiode is used to collect the sample's fluorescence. An electronic offset signal is used to reduce the effects of background fluorescence and the noises from the measurement system. An integrator integrates the difference between the output of the photodiode and the electronic offset signal over a given period of time. The resulting integral is then converted into digital domain for further processing and storage.

  11. A fully robust PARAFAC method for analyzing fluorescence data

    DEFF Research Database (Denmark)

    Engelen, Sanne; Frosch, Stina; Jørgensen, Bo

    2009-01-01

    Parallel factor analysis (PARAFAC) is a widespread method for modeling fluorescence data by means of an alternating least squares procedure. Consequently, the PARAFAC estimates are highly influenced by outlying excitation–emission landscapes (EEM) and element-wise outliers, like for example Raman......, there still exists no robust method for handling fluorescence data encountering both outlying EEM landscapes and scatter. In this paper, we present an iterative algorithm where the robust PARAFAC method and the scatter identification tool are alternately performed. A fully automated robust PARAFAC method...

  12. Laser Induced Fluorescence Spectroscopy of IrN

    Institute of Scientific and Technical Information of China (English)

    H. F. Pang; A. S. C. Cheung

    2009-01-01

    High resolution laser induced fluorescence spectra of IrN in the spectral region between 394and 520 nm were recorded using laser vaporization/reaction free jet expansion and laser induced fluorescence spectroscopy. Seven new vibronic transition bands were observed and analyzed. TwoΩ=1 and five Ω=0 new states were identified. Least squares fit of rotationally resolved transition lines yielded accurate molecular constants for the upper states. Spectra of isotopic molecules were observed, which provided confirmation for the vibrational assignment. Comparison of the observed electronic states of IrB, IrC, and IrN provides a good understanding of the chemical bonding of this group of molecules.

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

    Science.gov (United States)

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

    2015-05-01

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

  14. [Discrimination of Crude Oil Samples Using Laser-Induced Time-Resolved Fluorescence Spectroscopy].

    Science.gov (United States)

    Han, Xiao-shuang; Liu, De-qing; Luan, Xiao-ning; Guo, Jin-jia; Liu, Yong-xin; Zheng, Rong-er

    2016-02-01

    The Laser-induced fluorescence spectra combined with pattern recognition method has been widely applied in discrimination of different spilled oil, such as diesel, gasoline, and crude oil. However, traditional three-dimension fluorescence analysis method, which is not adapted to requirement of field detection, is limited to laboratory investigatio ns. The development of oil identification method for field detection is significant to quick response and operation of oil spill. In this paper, a new method based on laser-induced time-resolved fluorescence combined with support vector machine (SVM) model was introduced to discriminate crude oil samples. In this method, time-resolved spectra data was descended into two dimensions with selecting appropriate range in time and wavelength domains respectively to form a SVM data base. It is found that the classification accurate rate increased with an appropriate selection. With a selected range from 54 to 74 ns in time domain, the classification accurate rate has been increased from 83.3% (without selection) to 88.1%. With a selected wavelength range of 387.00~608.87 nm, the classification accurate rate of suspect oil was improved from 84% (without selection) to 100%. Since the detection delay of fluorescence lidar fluctuates due to wave and platform swing, the identification method with optimizing in both time and wavelength domains could offer a better flexibility for field applications. It is hoped that the developed method could provide some useful reference with data reduction for classification of suspect crude oil in the future development.

  15. A new method for measuring concentration of a fluorescent tracer in bubbly gas-liquid flows

    Science.gov (United States)

    Moghaddas, J. S.; Trägårdh, C.; Kovacs, T.; Östergren, K.

    2002-06-01

    A new experimental model, the two-tracer method (TTM), based on the planar laser-induced fluorescence technique (PLIF), is presented for the measurement of the local concentration of a fluorescent tracer in the liquid phase of a bubbly two-phase system. Light scattering and shading effects due to the bubbles were compensated for using the new model. The TTM results were found to give more accurate predictions of the local concentration than the normal PLIF method in a bubbly two-phase system.

  16. Metal-induced fluorescence properties of three-dimensionally ordered macroporous silver inverse opal platforms

    Science.gov (United States)

    Chae, Weon-Sik; Lee, Myung-Jin; Kim, Kisun; Hyun, Jerome K.; Jeon, Seokwoo

    2016-02-01

    This study examined the metal-induced fluorescence properties of three-dimensionally ordered macroporous silver inverse opal (IO) films. Electrochemically synthesized silver IO films with a micrometer cavity exhibited notable fluorescence enhancement at the silver frame, and a decrease in fluorescence lifetime. Numerical calculations supported the observations of a higher fluorescence efficiency at the frame than in the cavity.

  17. Fluorescence anion sensors based on combination of conformational restriction and photo-induced electron transfer

    Energy Technology Data Exchange (ETDEWEB)

    Liu, Ge, E-mail: liu_ge2008@163.com [Department of Chemistry, Chifeng University, Chifeng, 024000 (China); Shao, Jie, E-mail: njshao@live.cn [College of Chemistry, Chemical Engineering and Material Science, Soochow University, Suzhou, Jiangsu 215123 (China)

    2013-03-15

    Six fluorescence and colorimetric sensors (1-6) were designed and synthesized through a facile method. These compounds, whose structures were characterized by {sup 1}H NMR, elemental analyses and MS, could be easily prepared by simple synthesis method. In these sensor molecules, azo group and conjugated di-benzene unit acted as a chromophore and a fluorophore; respectively, and phenol moieties are anion binding sites. In dry DMSO solution, significant red shifts in UV-vis spectra with visible color changes were observed when the sensors were exposed to anions with strong base. Particularly, the simple compounds gave interesting fluorescence changes upon presence of anions, which probably resulted from electronic effects of the substituent. The sensors (1-3) with the nitro group exhibited 'turn-on' fluorescence responses to anions due to conformational restriction and 4-6 with absence of -NO{sub 2} group showed 'turn-off' fluorescence responses to anion owing to photo-induced electron transfer. - Highlights: Black-Right-Pointing-Pointer Six fluorescence and colorimetric anion sensors based on azo derivatives were showed. Black-Right-Pointing-Pointer Azo group and di-benzene unit acted as a chromophore and a fluorophore respectively. Black-Right-Pointing-Pointer Combination of conformational restriction and PET was exploited as fluorescence mechanism.

  18. Proton-induced x-ray fluorescence CT imaging.

    Science.gov (United States)

    Bazalova-Carter, Magdalena; Ahmad, Moiz; Matsuura, Taeko; Takao, Seishin; Matsuo, Yuto; Fahrig, Rebecca; Shirato, Hiroki; Umegaki, Kikuo; Xing, Lei

    2015-02-01

    To demonstrate the feasibility of proton-induced x-ray fluorescence CT (pXFCT) imaging of gold in a small animal sized object by means of experiments and Monte Carlo (MC) simulations. First, proton-induced gold x-ray fluorescence (pXRF) was measured as a function of gold concentration. Vials of 2.2 cm in diameter filled with 0%-5% Au solutions were irradiated with a 220 MeV proton beam and x-ray fluorescence induced by the interaction of protons, and Au was detected with a 3 × 3 mm(2) CdTe detector placed at 90° with respect to the incident proton beam at a distance of 45 cm from the vials. Second, a 7-cm diameter water phantom containing three 2.2-diameter vials with 3%-5% Au solutions was imaged with a 7-mm FWHM 220 MeV proton beam in a first generation CT scanning geometry. X-rays scattered perpendicular to the incident proton beam were acquired with the CdTe detector placed at 45 cm from the phantom positioned on a translation/rotation stage. Twenty one translational steps spaced by 3 mm at each of 36 projection angles spaced by 10° were acquired, and pXFCT images of the phantom were reconstructed with filtered back projection. A simplified geometry of the experimental data acquisition setup was modeled with the MC TOPAS code, and simulation results were compared to the experimental data. A linear relationship between gold pXRF and gold concentration was observed in both experimental and MC simulation data (R(2) > 0.99). All Au vials were apparent in the experimental and simulated pXFCT images. Specifically, the 3% Au vial was detectable in the experimental [contrast-to-noise ratio (CNR) = 5.8] and simulated (CNR = 11.5) pXFCT image. Due to fluorescence x-ray attenuation in the higher concentration vials, the 4% and 5% Au contrast were underestimated by 10% and 15%, respectively, in both the experimental and simulated pXFCT images. Proton-induced x-ray fluorescence CT imaging of 3%-5% gold solutions in a small animal sized water phantom has been demonstrated

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

    Science.gov (United States)

    Elliott, Drew; Scime, Earl; Short, Zachary

    2016-11-01

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

  20. Modified Methods for the Synthesis of Triazinyl Fluorescent Brightener Intermediates

    Directory of Open Access Journals (Sweden)

    Ali Parach

    2003-03-01

    Full Text Available The production of triazinyl fluorescent brightener intermediates in high yields is described. The method involves a simplified work-up for the preparation of 4-nitro-toluene-2-sulfonic acid and the use of diethylene glycol instead of water in the preparation of 4,4'-dinitrostilbene-2,2'-disulfonic acid.

  1. Fast multislice fluorescence molecular tomography using sparsity-inducing regularization.

    Science.gov (United States)

    Hejazi, Sedigheh Marjaneh; Sarkar, Saeed; Darezereshki, Ziba

    2016-02-01

    Fluorescence molecular tomography (FMT) is a rapidly growing imaging method that facilitates the recovery of small fluorescent targets within biological tissue. The major challenge facing the FMT reconstruction method is the ill-posed nature of the inverse problem. In order to overcome this problem, the acquisition of large FMT datasets and the utilization of a fast FMT reconstruction algorithm with sparsity regularization have been suggested recently. Therefore, the use of a joint L1/total-variation (TV) regularization as a means of solving the ill-posed FMT inverse problem is proposed. A comparative quantified analysis of regularization methods based on L1-norm and TV are performed using simulated datasets, and the results show that the fast composite splitting algorithm regularization method can ensure the accuracy and robustness of the FMT reconstruction. The feasibility of the proposed method is evaluated in an in vivo scenario for the subcutaneous implantation of a fluorescent-dye-filled capillary tube in a mouse, and also using hybrid FMT and x-ray computed tomography data. The results show that the proposed regularization overcomes the difficulties created by the ill-posed inverse problem.

  2. Fast multislice fluorescence molecular tomography using sparsity-inducing regularization

    Science.gov (United States)

    Hejazi, Sedigheh Marjaneh; Sarkar, Saeed; Darezereshki, Ziba

    2016-02-01

    Fluorescence molecular tomography (FMT) is a rapidly growing imaging method that facilitates the recovery of small fluorescent targets within biological tissue. The major challenge facing the FMT reconstruction method is the ill-posed nature of the inverse problem. In order to overcome this problem, the acquisition of large FMT datasets and the utilization of a fast FMT reconstruction algorithm with sparsity regularization have been suggested recently. Therefore, the use of a joint L1/total-variation (TV) regularization as a means of solving the ill-posed FMT inverse problem is proposed. A comparative quantified analysis of regularization methods based on L1-norm and TV are performed using simulated datasets, and the results show that the fast composite splitting algorithm regularization method can ensure the accuracy and robustness of the FMT reconstruction. The feasibility of the proposed method is evaluated in an in vivo scenario for the subcutaneous implantation of a fluorescent-dye-filled capillary tube in a mouse, and also using hybrid FMT and x-ray computed tomography data. The results show that the proposed regularization overcomes the difficulties created by the ill-posed inverse problem.

  3. An optimised method for correcting quenched fluorescence yield

    Directory of Open Access Journals (Sweden)

    L. Biermann

    2014-05-01

    Full Text Available 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.

  4. Temperature measurements of micro-droplets using pulsed 2-color laser-induced fluorescence with MDR-enhanced energy transfer

    Science.gov (United States)

    Palmer, Johannes; Reddemann, Manuel A.; Kirsch, Valeri; Kneer, Reinhold

    2016-12-01

    In this work, a new measurement system is presented for studying temperature of micro-droplets by pulsed 2-color laser-induced fluorescence. Pulsed fluorescence excitation allows motion blur suppression and thus simultaneous measurements of droplet size, velocity and temperature. However, high excitation intensities of pulsed lasers lead to morphology-dependent resonances inside micro-droplets, which are accompanied by disruptive stimulated emission. Investigations showed that stimulated emission can be avoided by enhanced energy transfer via an additional dye. The suitability and accuracy of the new pulsed method are verified on the basis of a spectroscopic analysis and comparison to continuously excited 2-color laser-induced fluorescence.

  5. Laser-induced fluorescence in high pressure solid propellant flames.

    Science.gov (United States)

    Edwards, T; Weaver, D P; Campbell, D H

    1987-09-01

    The application of laser-induced fluorescence (LIF) to the study of high pressure solid propellant flames is described. The distribution of the OH and CN radicals was determined in several solid propellant flames at pressures up to 3.5 MPa. The greatest difficulty in these measurements was the separation of the desired LIF signals from the large scattering at the laser wavelength from the very optically thick propellant flames. Raman experiments using 308-nm excitation were also attempted in the propellant flames but were unsuccessful due to LIF interferences from OH and NH.

  6. Blood perfusion and pH monitoring in organs by laser-induced fluorescence spectroscopy

    Science.gov (United States)

    Vari, Sandor G.; Papazoglou, Theodore G.; Pergadia, Vani R.; Stavridi, Marigo; Snyder, Wendy J.; Papaioannou, Thanassis; Duffy, J. T.; Weiss, Andrew B.; Thomas, Reem; Grundfest, Warren S.

    1994-01-01

    Sensitivity of laser-induced fluorescence spectroscopy (LIFS) in detecting a change in tissue pH, and blood perfusion was determined. Rabbits were anesthetized, paralyzed, and mechanically ventilated. The arterial and venous blood supplies of the kidney were isolated and ligated to alter the perfusion. The femoral artery was cannulated to extract samples for blood gas analysis. A 308-nm XeCl was used as an excitation source. A 600 micrometers core diameter fiber was used for fluorescence acquisition, and the spectra analyzed by an optical multichannel analyzer (EG & G, OMA III). the corresponding intensity ratio R equals INADH / ICOLL was used as an index for respiratory acidosis. Blood perfusion was assessed using the following algorithm: (IELAS minus ICOLL) divided by (INADH minus ICOLL). The intensity ratio linearly decreased with the reduction of blood perfusion. When we totally occluded the artery the ratio decreased tenfold when compared to the ratio of a fully perfused kidney. Results of monitoring blood acidosis by laser-induced fluorescence spectroscopy shows a significant trend between pH and intensity ratio. Since all the slopes were negative, there is an obvious significant correlation between the pH and NADH.COLLAGEN RATIO. Blue-light-induced fluorescence measurements and ratio fluorometry is a sensitive method for monitoring blood perfusion and acidity or alkalinity of an organ.

  7. Development of fluorescent methods for DNA methyltransferase assay

    Science.gov (United States)

    Li, Yueying; Zou, Xiaoran; Ma, Fei; Tang, Bo; Zhang, Chun-yang

    2017-03-01

    DNA methylation modified by DNA methyltransferase (MTase) plays an important role in regulating gene transcription, cell growth and proliferation. The aberrant DNA MTase activity may lead to a variety of human diseases including cancers. Therefore, accurate and sensitive detection of DNA MTase activity is crucial to biomedical research, clinical diagnostics and therapy. However, conventional DNA MTase assays often suffer from labor-intensive operations and time-consuming procedures. Alternatively, fluorescent methods have significant advantages of simplicity and high sensitivity, and have been widely applied for DNA MTase assay. In this review, we summarize the recent advances in the development of fluorescent methods for DNA MTase assay. These emerging methods include amplification-free and the amplification-assisted assays. Moreover, we discuss the challenges and future directions of this area.

  8. Laser-Induced Fluorescence Bronchoscopy for Detection and Localization of Early Lung Cancer

    Institute of Scientific and Technical Information of China (English)

    XIE Shusen; LI Buhong; LI Hui; ZHENG Wei; LU Zukang

    2001-01-01

    Experimenatal results on the development of a Laser-Induced Fluorescence Bronchoscopy(LIFB) for the detection and localization of early lung cancer are reported in this paper. The system utilizes fluorescence of photosensitizer drug to provide real time video imaging for the examined lung tissue. Color filters are used to differentiate signal from background and a computer image processing technique is also applied to subtract the background. Moreover, a pseudocolor contrast enhancement method was developed to enhance the fluorescence image displayed on the vidio monitor. Suspicious areas are identified by pseudocolor image to guide biopsy, and several clinical trials show that sensitivity and contrast capability of the system should permit the detection and localization of early lung cancer.

  9. An adaptive Tikhonov regularization method for fluorescence molecular tomography.

    Science.gov (United States)

    Cao, Xu; Zhang, Bin; Wang, Xin; Liu, Fei; Liu, Ke; Luo, Jianwen; Bai, Jing

    2013-08-01

    The high degree of absorption and scattering of photons propagating through biological tissues makes fluorescence molecular tomography (FMT) reconstruction a severe ill-posed problem and the reconstructed result is susceptible to noise in the measurements. To obtain a reasonable solution, Tikhonov regularization (TR) is generally employed to solve the inverse problem of FMT. However, with a fixed regularization parameter, the Tikhonov solutions suffer from low resolution. In this work, an adaptive Tikhonov regularization (ATR) method is presented. Considering that large regularization parameters can smoothen the solution with low spatial resolution, while small regularization parameters can sharpen the solution with high level of noise, the ATR method adaptively updates the spatially varying regularization parameters during the iteration process and uses them to penalize the solutions. The ATR method can adequately sharpen the feasible region with fluorescent probes and smoothen the region without fluorescent probes resorting to no complementary priori information. Phantom experiments are performed to verify the feasibility of the proposed method. The results demonstrate that the proposed method can improve the spatial resolution and reduce the noise of FMT reconstruction at the same time.

  10. A laser-induced fluorescence measurement technique for obtaining neutral hydrogen densities in plasmas

    Energy Technology Data Exchange (ETDEWEB)

    Yao, X.Z.; Yang, T.F.; Chang-Diaz, F.R. [Plasma Fusion Center, Massachusetts Institute of Technology, Cambridge, Massachusetts 02139 (United States)

    1995-09-01

    The resonance fluorescence of neutral hydrogen illuminated by {ital H}{sub {alpha}} radiation has been used as a technique for the spatially and temporally resolved density measurements of neutral hydrogen in high temperature plasmas, such as in the tokamak and magnetic mirror plasma fusion devices. The fluorescence signal, usually very weak and buried in the background of stray laser light and {ital H}{sub {alpha}} emission, is very difficult to extract and its measurements are inaccurate. This paper discusses the improvement of the signal extraction using two optical path laser-induced fluorescence (LIF) methods. One optical path carries the fluorescence signal and the background (the stray laser light and {ital H}{sub {alpha}} emission), whereas the other path carries only the background signal. By combining these two signals, a clean fluorescence signal can be isolated by subtracting out the background using a differential amplifier. The measurement is obtained instantaneously from these two signals which are taken simultaneously in one pulse rather than being extracted from two separate spectra taken in two sequential pulses (double pulses). This method, therefore, makes a significant improvement on the double pulse technique in terms of the accuracy of the measurement and the time resolution. Using this LIF technique the measurement of the neutral density profile in the exhaust of a tandem mirror plasma propulsion device is obtained and presented. {copyright} {ital 1995} {ital American} {ital Institute} {ital of} {ital Physics}.

  11. Intermolecular-charge-transfer-induced fluorescence quenching in protic solvent

    Science.gov (United States)

    Lin, Tao; Liu, Xiaojun; Lou, Zhidong; Hou, Yanbing; Teng, Feng

    2016-11-01

    The fluorescence quenching of fluorenone in protic solvent has been extensively investigated, and the intermolecular hydrogen bond was found to play a crucial role. Unfortunately, the mechanism at atomic level is still not clear. In the present work, we theoretically put forward the charge transfer along the hydrogen bond in the excited states. The vertical excitation energies of the fluorenone-methanol complex as well as the potential energy profiles and surfaces of the vertical excited states and charge transfer states were calculated by using the ab initio electronic-structure methods. The photochemical reactions occurring in the diverse charge transfer states were compared and their decisiveness to the fluorescence quenching was discussed in the paper.

  12. Canopy Level Solar Induced Fluorescence for Vegetation in Controlled Experiments

    Science.gov (United States)

    Middleton, E. M.; Corp, L. A.; Campbell, P. K. Entcheva

    2007-01-01

    Solar induced chlorophyll fluorescence (SIF) was retrieved from high resolution reflectance spectra acquired one meter above saplings of three deciduous tree species during springtime (three weeks after leaf flush) and in late summer when foliage was mature. SIF was determined by application of the Fraunhofer Line Depth (FLD) Principal to above-canopy spectra acquired with an Analytical Spectral Devices (ASD) Fieldspec spectroradiometer (3.2 nm resolution with 1.2 nm sampling interval). SIF retrievals were made at the two atmospheric oxygen (O2) absorption features that occur in the chlorophyll fluorescence (ChlF) region (660 -780 nm). These telluric features are 02V, the broader and deeper feature centered at 760 nm, but located on the shoulder of the far-red ChlF peak at 740 nm; and 023, a narrow feature centered at 688 nm that is positioned near the red ChlF peak at 685 nm. Supporting, coincident leaf level fluorescence, reflectance, photochemical and other measurements were also made. At the leaf level, these measurements included in situ photosynthetic capacity (Pmax) and light adapted total chlorophyll fluorescence (Fs') collected at steady state under high light and controlled chamber conditions (e.g., temperature, PAR, humidity, and COz); optical properties (reflectance, transmittance, absorptance); chlorophyll and carotenoid content; specific leaf mass; carbon (C) and nitrogen (N) content; fluorescence emission spectra at multiple excitation wavelengths; the ChlF contribution to red (R) and far-red (FR) reflectance; fluorescence imagery; and fluorescence excitation-emission matrices (EEMs). The tree species examined were tulip poplar (Liriodendron tulipifera L.), red maple (Acer rubrum L.), and sweetgum (Liquidambar styraczflua L.), and each had been provided four levels of N augmentation (0, 19, 37, and 75 kg Nhectare seasonally) to simulate atmospheric deposition from air pollution. Whole-plant SIF measurements of these species were compared with SIF

  13. Contrast Induced by a Static Magnetic Field for Improved Detection in Nanodiamond Fluorescence Microscopy

    Science.gov (United States)

    Singam, Shashi K. R.; Motylewski, Jaroslaw; Monaco, Antonina; Gjorgievska, Elena; Bourgeois, Emilie; Nesládek, Milos; Giugliano, Michele; Goovaerts, Etienne

    2016-12-01

    Diamond nanoparticles with negatively charged nitrogen-vacancy (NV) centers are highly efficient nonblinking emitters that exhibit spin-dependent intensity. An attractive application of these emitters is background-free fluorescence microscopy exploiting the fluorescence quenching induced either by resonant microwaves (RMWs) or by an applied static magnetic field (SMF). Here, we compare RMW- and SMF-induced contrast measurements over a wide range of optical excitation rates for fluorescent nanodiamonds (FNDs) and for NV centers shallowly buried under the (100)-oriented surface of a diamond single crystal (SC). Contrast levels are found to be systematically lower in the FNDs than in the SC. At low excitation rates, the RMW contrast initially rises to a maximum (up to 7% in FNDs and 13% in the SC) but then decreases steadily at higher intensities. Conversely, the SMF contrast increases from approximately 12% at low excitation rates to high values of 20% and 38% for the FNDs and SC, respectively. These observations are well described in a rate-equations model for the charged NV defect using parameters in good agreement with the literature. The SMF approach yields higher induced contrast in image collection under commonly applied optical excitation. Unlike the RMW method, there is no thermal load exerted on the aqueous media in biological samples in the SMF approach. We demonstrate imaging by SMF-induced contrast in neuronal cultures incorporating FNDs (i) in a setup for patch-clamp experiments in parallel with differential-interference-contrast microscopy, (ii) after a commonly used staining procedure as an illustration of the high selectivity against background fluorescence, and (iii) in a confocal fluorescence microscope in combination with bright-field microscopy.

  14. Formation of fluorescent polydopamine dots from hydroxyl radical-induced degradation of polydopamine nanoparticles.

    Science.gov (United States)

    Lin, Jia-Hui; Yu, Cheng-Ju; Yang, Ya-Chun; Tseng, Wei-Lung

    2015-06-21

    This study describes the synthesis of fluorescent polydopamine dots (PDs) through hydroxyl radical-induced degradation of polydopamine nanoparticles. The decomposition of polydopamine nanoparticles to fluorescent PDs was confirmed using transmission electron microscopy and dark-field microscopy. The analysis of PDs by using laser desorption/ionization time-of-flight mass spectrometry revealed that the PDs consisted of dopamine, 5,6-dihydroxyindole, and trihydroxyindole units. Oligomerization and self-assembly of these units produced a broad adsorption band, resulting in an excitation-wavelength-dependent emission behavior. The maximal fluorescence of PDs appeared at 440 nm with a quantum yield of 1.2%. The coordination between the catechol groups of PDs and ferric ions (Fe(3+)) quenched the fluorescence of PDs; the limit of detection at a signal-to-noise ratio of 3 for Fe(3+) was determined to be 0.3 μM. The presence of pyrophosphate switched on the fluorescence of the PD-Fe(3+) complexes. Compared to the other reported methods for sensing Fe(3+), PDs provided simple, low-cost, and reusable detection of Fe(3+).

  15. Serum Protein Profile Study of Clinical Samples Using High Performance Liquid Chromatography-Laser Induced Fluorescence

    DEFF Research Database (Denmark)

    Karemore, Gopal Raghunath; Ukendt, Sujatha; Rai, Lavanya

    2009-01-01

    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for estab......The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested...... for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions....

  16. Quantitative laser-induced fluorescence measurements of nitric oxide in a heavy-duty Diesel engine

    NARCIS (Netherlands)

    Verbiezen, K.; Klein-Douwel, R. J. H.; van Viet, A. P.; Donkerbroek, A. J.; Meerts, W. L.; Dam, N. J.; ter Meulen, J. J.

    2007-01-01

    We present quantitative, in-cylinder, UV-laser-induced fluorescence measurements of nitric oxide in a heavy-duty Diesel engine. Processing of the raw fluorescence signals includes a detailed correction, based on additional measurements, for the effect of laser beam and fluorescence attenuation, and

  17. Imaging magnetic scalar potentials by laser-induced fluorescence from bright and dark atoms

    CERN Document Server

    Fescenko, Ilja

    2014-01-01

    We present a spectroscopic method for mapping two-dimensional distributions of magnetic field strengths (magnetic scalar potential lines) using CCD recordings of the fluorescence patterns emitted by spin-polarized Cs vapor in a buffer gas exposed to inhomogeneous magnetic fields. The method relies on the position-selective destruction of spin polarization by magnetic resonances induced by multi-component oscillating magnetic fields, such that magnetic potential lines can directly be detected by the CCD camera. We also present a generic algebraic model allowing the calculation of the fluorescence patterns and find excellent agreement with the experimental observations for three distinct inhomogeneous field topologies. The spatial resolution obtained with these proof-of-principle experiments is on the order of 1 mm. A substantial increase of spatial and magnetic field resolution is expected by deploying the method in a magnetically shielded environment.

  18. Initial operation of the LEDA beam-induced fluorescence diagnostic

    Science.gov (United States)

    Kamperschroer, James H.; Gilpatrick, J. Douglas; Gurd, Pamela A.; Madsen, David W.; Martinez, Derwin G.; O'Hara, James F.; Sage, Joan; Schaefer, Timothy L.; Shurter, R. Bradford; Stettler, Matthew W.

    2000-11-01

    A diagnostic based on beam-induced fluorescence has been developed and used to examine the expanded beam in the High-Energy Beam Transport (HEBT) section of the Low Energy Demonstration Accelerator (LEDA). The system consists of a camera, a gas injector, a spectrometer, and a control system. Gas is injected to provide a medium for the beam to excite, the camera captures the resulting image of the fluorescing gas, and the spectrometer measures the spectrum of the emitted light. EPICS was used to control the camera and acquire and store images. Data analysis is presently being performed offline. A Kodak DCS420m professional CCD camera is the primary component of the optical system. InterScience, Inc. modified the camera with the addition of a gain of 4000 image intensifier, thereby producing an intensified camera with a sensitivity of ˜0.5 milli-lux. Light is gathered with a 1″ format, 16-160 mm, Computar zoom lens. This lens is attached to the camera via a Century Precision Optics relay lens. Images obtained using only hydrogen from the beam stop exhibited features not yet understood. Images with good signal-to-noise ratio were obtained with the injection of sufficient nitrogen to raise the HEBT pressure to 2-8×10-6 torr. Two strong nitrogen lines, believed to be of the first negative group of N2+, were identified at 391 and 428 nm.

  19. Containerless study of metal evaporation by laser induced fluorescence

    Science.gov (United States)

    Schiffman, Robert A.; Nordine, Paul C.

    1987-01-01

    Laser induced fluorescence (LIF) detection of atomic vapors was used to study evaporation from electromagnetically levitated and CW CO2 laser-heated molybdenum spheres and resistively-heated tungsten filaments. Electromagnetic (EM) levitation in combination with laser heating of tungsten, zirconium, and aluminum specimens was also investigated. LIF intensity vs temperature data were obtained for molybdenum atoms and six electronic states of atomic tungsten, at temperatures up to the melting point of each metal. The detected fraction of the emitted radiation was reduced by self-absorption effects at the higher experimental temperatures. Vaporization enthalpies derived from data for which less than half the LIF intensity was self-absorbed were -636 + or - 24 kJ/g-mol for Mo and 831 + or - 32 kJ/g-mol for W. Space-based applications of EM levitation in combination with radiative heating are discussed.

  20. Clinical detection studies of Barrett's metaplasia and oesophageal adenocarcinoma by means of laser-induced fluorescence

    Science.gov (United States)

    Nilsson, Annika M.; von Holstein, Christer S.; Andersson-Engels, Stefan; Willen, Roger; Walther, Bruno; Svanberg, Katarina

    1995-12-01

    Five patients with Barrett's metaplastic epithelium were investigated by means of laser- induced fluorescence after low-dose i.v. injection (0.35 mg/kg b.w.) of PhotofrinR in connection with endoscopy procedures. The excitation wavelength was 405 nm. Recorded fluorescence spectra were evaluated by forming ratios with the photosensitizer fluorescence as numerator and the autofluorescence as denominator. Two patients had no evidence of malignancy and their fluorescence ratios were consequently rather small, whereas the other three patients had adenocarcinoma and showed considerably higher ratios. The results indicate that laser-induced fluorescence can be used as an aid in detecting malignant transformations in Barrett's metaplasia.

  1. Cyanine-based probe\\tag-peptide pair fluorescence protein imaging and fluorescence protein imaging methods

    Science.gov (United States)

    Mayer-Cumblidge, M. Uljana; Cao, Haishi

    2013-01-15

    A molecular probe comprises two arsenic atoms and at least one cyanine based moiety. A method of producing a molecular probe includes providing a molecule having a first formula, treating the molecule with HgOAc, and subsequently transmetallizing with AsCl.sub.3. The As is liganded to ethanedithiol to produce a probe having a second formula. A method of labeling a peptide includes providing a peptide comprising a tag sequence and contacting the peptide with a biarsenical molecular probe. A complex is formed comprising the tag sequence and the molecular probe. A method of studying a peptide includes providing a mixture containing a peptide comprising a peptide tag sequence, adding a biarsenical probe to the mixture, and monitoring the fluorescence of the mixture.

  2. A Quantitative Method for Microtubule Analysis in Fluorescence Images.

    Science.gov (United States)

    Lan, Xiaodong; Li, Lingfei; Hu, Jiongyu; Zhang, Qiong; Dang, Yongming; Huang, Yuesheng

    2015-12-01

    Microtubule analysis is of significant value for a better understanding of normal and pathological cellular processes. Although immunofluorescence microscopic techniques have proven useful in the study of microtubules, comparative results commonly rely on a descriptive and subjective visual analysis. We developed an objective and quantitative method based on image processing and analysis of fluorescently labeled microtubular patterns in cultured cells. We used a multi-parameter approach by analyzing four quantifiable characteristics to compose our quantitative feature set. Then we interpreted specific changes in the parameters and revealed the contribution of each feature set using principal component analysis. In addition, we verified that different treatment groups could be clearly discriminated using principal components of the multi-parameter model. High predictive accuracy of four commonly used multi-classification methods confirmed our method. These results demonstrated the effectiveness and efficiency of our method in the analysis of microtubules in fluorescence images. Application of the analytical methods presented here provides information concerning the organization and modification of microtubules, and could aid in the further understanding of structural and functional aspects of microtubules under normal and pathological conditions.

  3. Laser-Induced Fluorescence in Gaseous [I[subscript]2] Excited with a Green Laser Pointer

    Science.gov (United States)

    Tellinghuisen, Joel

    2007-01-01

    A green laser pointer could be used in a flashy demonstration of laser-induced fluorescence in the gas phase by directing the beam of the laser through a cell containing [I[subscript]2] at its room temperature vapor pressure. The experiment could be used to provide valuable insight into the requirements for laser-induced fluorescence (LIF) and the…

  4. Correction of depth-induced spherical aberration for deep observation using two-photon excitation fluorescence microscopy with spatial light modulator.

    Science.gov (United States)

    Matsumoto, Naoya; Inoue, Takashi; Matsumoto, Akiyuki; Okazaki, Shigetoshi

    2015-07-01

    We demonstrate fluorescence imaging with high fluorescence intensity and depth resolution in which depth-induced spherical aberration (SA) caused by refractive-index mismatch between the medium and biological sample is corrected. To reduce the impact of SA, we incorporate a spatial light modulator into a two-photon excitation fluorescence microscope. Consequently, when fluorescent beads in epoxy resin were observed with this method of SA correction, the fluorescence signal of the observed images was ∼27 times higher and extension in the direction of the optical axes was ∼6.5 times shorter at a depth of ∼890 μm. Thus, the proposed method increases the depth observable at high resolution. Further, our results show that the method improved the fluorescence intensity of images of the fluorescent beads and the structure of a biological sample.

  5. Spectrally resolved laser-induced fluorescence for bioaerosols standoff detection

    Science.gov (United States)

    Buteau, Sylvie; Stadnyk, Laurie; Rowsell, Susan; Simard, Jean-Robert; Ho, Jim; Déry, Bernard; McFee, John

    2007-09-01

    An efficient standoff biological warfare detection capability could become an important asset for both defence and security communities based on the increasing biological threat and the limits of the presently existing protection systems. Defence R&D Canada (DRDC) has developed, by the end of the 90s, a standoff bioaerosol sensor prototype based on intensified range-gated spectrometric detection of Laser Induced Fluorescence (LIF). This LIDAR system named SINBAHD monitors the spectrally resolved LIF originating from inelastic interactions with bioaerosols present in atmospheric cells customizable in size and in range. SINBAHD has demonstrated the capability of near real-time detection and classification of bioaerosolized threats at multi-kilometre ranges. In spring 2005, DRDC has initiated the BioSense demonstration project, which combines the SINBAHD technology with a geo-referenced Near InfraRed (NIR) LIDAR cloud mapper. SINBAHD is now being used to acquire more signatures to add in the spectral library and also to optimize and test the new BioSense algorithm strategy. In September 2006, SINBAHD has participated in a two-week trial held at DRDC-Suffield where different open-air wet releases of live and killed bioagent simulants, growth media and obscurants were performed. An autoclave killing procedure was performed on two biological materials (Bacillus subtilis var globigii or BG, and Bacillus thuringiensis or Bt) before being aerosolized, disseminated and spectrally characterized with SINBAHD. The obtained results showed no significant impact of this killing process on their normalised spectral signature in comparison with their live counterparts. Correlation between the detection signals from SINBAHD, an array of slit samplers and a FLuorescent Aerosol Particle Sensor (C-FLAPS) was obtained and SINBAHD's sensitivity could then be estimated. At the 2006 trial, a detection limit of a few tens of Agent Containing Particles per Liter of Air (ACPLA) was obtained

  6. Fluorescence spectroscopic detection of virus-induced atherosclerosis

    Science.gov (United States)

    Yan, Wei-dong; Perk, Masis; Nation, Patric N.; Power, Robert F.; Liu, Liying; Jiang, Xiuyan; Lucas, Alexandra

    1994-07-01

    Laser-induced fluorescence (LF) has been developed as a diagnostic tool for the detection of atherosclerosis. We have examined the use of LF for the identification of accelerated atherosclerotic plaque growth induced by Marek's Disease Virus (MDV) infection in White Leghorn rooster chicks (R) as well as plaque regression after treatment. Twenty-eight newborn R were infected with 12,000 cfu of MDV. Twelve parallel control R had saline injection. LF spectra were recorded from the arteries in vitro with a CeramOptec laser angioplasty catheter during 308 nm XeCl excimer laser excitation. Significant differences were detected at 440 to 475, 525, 550, 600, and 650 nm in MDV-R (p<0.05). In a subsequent study, 60 R were infected with 5,000 cfu of MDV, and were then treated with either Pravastatin (PRV) or placebo at 3 months post infection. These PRV-R were followed for 6 months to detect changes in atherosclerotic plaque development. PRV reduced intimal proliferation produced by MDV infection on histological examination (PRV-R 128.0+/- 44.0 micrometers , placebo-R 412.2+/- 91.5 micrometers , pequals0.007). MDV infected, PRV treated R were examined for LF changes that correlated with decreased atherosclerosis. There was an associated significant increase in LF intensity in PRV-R at 405 to 425 nm (p<0.001). In conclusion, LF can detect intimal proliferation in virus- induced atherosclerosis and atherosclerotic plaque regression after PRV therapy.

  7. The use of one- and two- photon induced fluorescence spectroscopy for the optical characterization of carcinogenic aflatoxins

    Science.gov (United States)

    Smeesters, L.; Meulebroeck, W.; Raeymaekers, S.; Thienpont, H.

    2014-09-01

    Carcinogenic and toxic contaminants in food and feed products are nowadays mostly detected by destructive, time-consuming chemical analyses, like HPLC and LC-MS/MS methods. However, as a consequence of the severe and growing regulations on food products by the European Union, there arose an increased demand for the ultra-fast, high-sensitive and non-destructive detection of contaminants in food and feed products. Therefore, we have investigated fluorescence spectroscopy for the characterization of carcinogenic aflatoxins. With the use of a tunable titanium-sapphire laser in combination with second and third harmonic wavelength generation, both one- and two-photon induced fluorescence excitation wavelengths could be generated using the same setup. We characterized and compared the one- and two-photon induced fluorescence spectra of pure aflatoxin powder, after excitation with 365nm and 730nm respectively. Moreover, we investigated the absolute fluorescence intensity as function of the excitation power density. Afterwards, we applied our characterization setup to the detection of aflatoxins in maize grains. The fluorescence spectra of both healthy and contaminated maize samples were experimentally characterized. In addition to the fluorescence spectrum of the pure aflatoxin, we observed an unwanted influence of the intrinsic fluorescence of the maize. Depending on the excitation wavelength, a varying contrast between the fluorescence spectra of the healthy and contaminated samples was obtained. After a comparison of the measured fluorescence signals, a detection criterion for the optical identification of the contaminated maize samples could be defined. As a result, this illustrates the use of fluorescence spectroscopy as a valuable tool for the non-destructive, real-time and high-sensitive detection of aflatoxins in maize.

  8. Chemometric treatment of multimode laser-induced fluorescence (LIF) data of fuel-spiked soils

    Science.gov (United States)

    Van Benthem, Mark H.; Mitchell, Ben C.; Gillispie, Gregory D.; St. Germain, Randy W.

    1996-11-01

    Field screening of fuel-contaminated soils using laser- induced fluorescence is a cost effective and timely method of characterizing contaminated sites. Data collected with laser-based screening tools are often extensive and difficult to interpret. Pattern recognition algorithms can be utilized to enable less highly trained personnel to identify contaminants. In this work, fluorescence intensity of various hydrocarbon fuels deposited on various soil types was measured as a function of emission wavelength and decay time, generating wavelength-time matrices. The data were arranged into a three mode array and subjected to trilinear decomposition (TLD). The results of the TLD were then utilized in pattern recognition schemes, specifically, linear discrimination and classification and hierarchical cluster analysis. Classification rates and clustering results indicate that these techniques can be very valuable tools in site characterization.

  9. Laser-induced fluorescence measurements on CdSe quantum dots

    Directory of Open Access Journals (Sweden)

    Zoltan Győri

    2010-03-01

    Full Text Available In this paper, we report on photoluminescence decay measurements on CdSe quantum dots (QDs as a function of size in the diameter range of 2.1 to 3.5 nm. The nanoparticles were synthesized by the kinetic growth method from CdO and elemental Se precursors. We studied the effects of growth time on the diameter, emission spectrum and the fluorescence lifetime of the synthesized QDs. The decay time measurements were performed using single shot time-resolved laser-induced fluorescence techniques using a Nd:YAG laser system. Two different decay times were measured on each CdSe sample, a fast one and a relatively slow one. The slow decay was found to be size dependent whereas the fast one was independent of the QD diameter.

  10. Native Fluorescence Detection Methods, Devices, and Systems for Organic Compounds

    Science.gov (United States)

    Hug, William F. (Inventor); Bhartia, Rohit (Inventor); Reid, Ray D. (Inventor); Lane, Arthur L. (Inventor)

    2016-01-01

    Naphthalene, benzene, toluene, xylene, and other volatile organic compounds VOCs have been identified as serious health hazards. Embodiments of the invention are directed to methods and apparatus for near-real-time in-situ detection and accumulated dose measurement of exposure to naphthalene vapor and other hazardous gaseous VOCs. The methods and apparatus employ excitation of fluorophors native or endogenous to compounds of interest using light sources emitting in the ultraviolet below 300 nm and measurement of native fluorescence emissions in distinct wavebands above the excitation wavelength. The apparatus of some embodiments are cell-phone-sized sensor/dosimeter "badges" to be worn by personnel potentially exposed to hazardous VOCs. The badge sensor of some embodiments provides both real time detection and data logging of exposure to naphthalene or other VOCs of interest from which both instantaneous and accumulated dose can be determined.

  11. Formalin-induced fluorescence reveals cell shape and morphology in biological tissue samples.

    Directory of Open Access Journals (Sweden)

    Ulrich Leischner

    Full Text Available Ultramicroscopy is a powerful tool to reveal detailed three-dimensional structures of large microscopical objects. Using high magnification, we observed that formalin induces fluorescence more in extra-cellular space and stains cellular structures negatively, rendering cells as dark objects in front of a bright background. Here, we show this effect on a three-dimensional image stack of a hippocampus sample, focusing on the CA1 region. This method, called FIF-Ultramicroscopy, allows for the three-dimensional observation of cellular structures in various tissue types without complicated staining techniques.

  12. Nanoantenna array-induced fluorescence enhancement and reduced lifetimes

    DEFF Research Database (Denmark)

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

    2008-01-01

    Enhanced fluorescence is observed from dye molecules interacting with optical nanoantenna arrays. Elliptical gold dimers form individual nanoantennae with tunable plasmon resonances depending upon the geometry of the two particles and the size of the gap between them. A fluorescent dye, Rhodamine...

  13. Nonnegative matrix factorization: a blind spectra separation method for in vivo fluorescent optical imaging.

    Science.gov (United States)

    Montcuquet, Anne-Sophie; Hervé, Lionel; Navarro, Fabrice; Dinten, Jean-Marc; Mars, Jérôme I

    2010-01-01

    Fluorescence imaging in diffusive media is an emerging imaging modality for medical applications that uses injected fluorescent markers that bind to specific targets, e.g., carcinoma. The region of interest is illuminated with near-IR light and the emitted back fluorescence is analyzed to localize the fluorescence sources. To investigate a thick medium, as the fluorescence signal decreases with the light travel distance, any disturbing signal, such as biological tissues intrinsic fluorescence (called autofluorescence) is a limiting factor. Several specific markers may also be simultaneously injected to bind to different molecules, and one may want to isolate each specific fluorescent signal from the others. To remove the unwanted fluorescence contributions or separate different specific markers, a spectroscopic approach is explored. The nonnegative matrix factorization (NMF) is the blind positive source separation method we chose. We run an original regularized NMF algorithm we developed on experimental data, and successfully obtain separated in vivo fluorescence spectra.

  14. Development of a UV laser-induced fluorescence lidar for monitoring blue-green algae in Lake Suwa.

    Science.gov (United States)

    Saito, Yasunori; Takano, Kengo; Kobayashi, Fumitoshi; Kobayashi, Kazuki; Park, Ho-Dong

    2014-10-20

    We developed a UV (355 nm) laser-induced fluorescence (LIF) lidar for monitoring the real-time status of blue-green algae. Since the fluorescence spectrum of blue-green algae excited by 355 nm showed the specific fluorescence at 650 nm, the lidar was designed to be able to detect the 650 nm fluorescence as a surveillance method for the algae. The usefulness was confirmed by observation at Lake Suwa over four years (2005-2008). The detection limit of the LIF lidar was 16.65 mg/L for the blue-green algae, which is the range of concentrations in the safe level set by the World Health Organization.

  15. Laser-induced fluorescence and optical reflection spectra of Japanese natural dyes on silk

    OpenAIRE

    Miyoshi, Tadaki; Matsuda, Yasunori

    1987-01-01

    Fluorescence spectra under nitrogen-laser excitation were measured for silk cloth dyed with Japanese natural dyes. An identification of the dyes on silk was carried out using a laser-induced fluorescence (LIF) technique since dyed cloth has a characteristic fluorescence spectra. Moreover, it is possible to identify dyes on faded cloth and on cloth prepared by a combination dyeing using two kinds of dyes. The LIF technique can identify dyes on cloth which is difficult to identify using the ref...

  16. Characterisation of estuarine intertidal macroalgae by laser-induced fluorescence

    DEFF Research Database (Denmark)

    Gameiro, Carla; Utkin, Andrei B.; Sousa Dias Cartaxana, Paulo Jorge

    2015-01-01

    spectra were determined by differences in the main fluorescing pigments: phycoerythrin, phycocyanin and chlorophyll a (Chl a). In the green and brown macroalgae groups, the relative significance of the two emission maxima seems to be related to the thickness of the photosynthetic layer. In thick...... macroalgae, like Codium tomentosum or Fucus vesiculosus, the contribution of the far-red emission fluorescence peak was more significant, most probably due to re-absorption of the emitted red Chl a fluorescence within the dense photosynthetic layer. Similarly, an increase in the number of layers of the thin...

  17. Laser induced fluorescence measurements of the mixing of fuel oil with air

    Energy Technology Data Exchange (ETDEWEB)

    Arnold, A.; Bombach, R.; Hubschmid, W.; Kaeppeli, B. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1999-08-01

    We report on measurements of the mixing of fuel oil with air at atmospheric pressure in an industrial premixed gas turbine burner. The concentration of the vaporized fuel oil was measured with laser induced fluorescence. We reason that the fuel oil concentration can be considered with good accuracy as proportional to the fluorescence intensity. (author) 6 fig., 3 refs.

  18. Applying fluorescence correlation spectroscopy to investigate peptide-induced membrane disruption

    DEFF Research Database (Denmark)

    Kristensen, Kasper; Henriksen, Jonas Rosager; Andresen, Thomas Lars

    2017-01-01

    to quantify leakage of fluorescent molecules of different sizes from large unilamellar lipid vesicles, thereby providing a tool for estimating the size of peptide-induced membrane disruptions. If fluorescently labeled lipids are incorporated into the membranes of the vesicles, FCS can also be used to obtain...

  19. Recognition of edible oil by using BP neural network and laser induced fluorescence spectrum

    Science.gov (United States)

    Mu, Tao-tao; Chen, Si-ying; Zhang, Yin-chao; Guo, Pan; Chen, He; Zhang, Hong-yan; Liu, Xiao-hua; Wang, Yuan; Bu, Zhi-chao

    2013-09-01

    In order to accomplish recognition of the different edible oil we set up a laser induced fluorescence spectrum system in the laboratory based on Laser induced fluorescence spectrum technology, and then collect the fluorescence spectrum of different edible oil by using that system. Based on this, we set up a fluorescence spectrum database of different cooking oil. It is clear that there are three main peak position of different edible oil from fluorescence spectrum chart. Although the peak positions of all cooking oil were almost the same, the relative intensity of different edible oils was totally different. So it could easily accomplish that oil recognition could take advantage of the difference of relative intensity. Feature invariants were extracted from the spectrum data, which were chosen from the fluorescence spectrum database randomly, before distinguishing different cooking oil. Then back propagation (BP) neural network was established and trained by the chosen data from the spectrum database. On that basis real experiment data was identified by BP neural network. It was found that the overall recognition rate could reach as high as 83.2%. Experiments showed that the laser induced fluorescence spectrum of different cooking oil was very different from each other, which could be used to accomplish the oil recognition. Laser induced fluorescence spectrum technology, combined BP neural network,was fast, high sensitivity, non-contact, and high recognition rate. It could become a new technique to accomplish the edible oil recognition and quality detection.

  20. [Studies on laticifers and milk of greater celandine (Chelidonium majus L.) with fluorescence imaging and fluorescence spectroscopic methods].

    Science.gov (United States)

    Póczi, Dorottya; Böddi, Béla

    2010-01-01

    Using fluorescence imaging and fluorescence spectroscopic methods, the localisation of the laticifers and the native spectral properties of the milk were studied in various organs of greater celandine (Chelidonium majus L.). Direct measurements on tissue pieces (without the extraction and the separation of the components) provided information about the complexity of the milk and the various ratios of the alkaloid contents in the tissues. Whole plant were studied in a gel documentation system using ultraviolet light source, while the localisation of the laticifers was observed along the leaf veins in fluorescence microscope, using blue excitation light. Measuring different tissue pieces, fluorescence spectroscopic studies showed that the greater celandine alkaloids have emission bands at 469, 530-531, 553, 572-575 and 592 nm and excitation bands at 365, 370, 386 is 400 nm. These results give a possibility for conclusions about the alkaloid contents and composition or ratios of the alkaloid components in various tissue pieces directly, via comparisons with alkaloid standards.

  1. Ion chromatographic method for the simultaneous determination of nitrite and nitrate by post-column indirect fluorescence detection.

    Science.gov (United States)

    Stalikas, Constantine D; Konidari, Constantina N; Nanos, Christos G

    2003-06-20

    This short paper highlights the suitability of ion chromatography with post-column indirect fluorescence detection to determine simultaneously nitrite and nitrate based on the quenching of tryptophan native fluorescence. The method uses an enhanced fluorescence mobile phase containing tryptophan and detects the suppression of fluorescence of the mobile phase due to the elution of the target ions. The phenomenon of fluorescence quenching of tryptophan is highly induced by the presence of phosphate ions. The quenched fluorescence intensity exhibits concentration dependence in the range 1-25 mg/l and 3-65 mg/l for nitrite and nitrate, respectively. The relative standard deviation for five replicates of a standard solution containing a mixture of 5 mg/l of nitrite and 10 mg/l of nitrate lies around 2.8%. This simple coupling technique results in a relatively sensitive, fast, and accurate method, allowing for both qualitative and quantitative analysis of nitrite and nitrate. The method can easily be implemented to real samples such as foodstuffs, fertilizers and soils and is proven to be precise and accurate when compared with reference methods.

  2. The yield of air fluorescence induced by electrons

    CERN Document Server

    Arqueros, F; Castellanos, A; Ortiz, M; Rosado, J

    2006-01-01

    The fluorescence yield for dry air and pure nitrogen excited by electrons is calculated using a combination of well-established molecular properties and experimental data of the involved cross sections. Particular attention has been paid to the role of secondary electrons from ionization processes. At high pressure and high energy, observed fluorescence turns out to be proportional to the ionization cross section which follows the Born-Bethe law. Predictions on fluorescence yields in a very wide interval of electron energies (eV - GeV) and pressures (1 and 1013 hPa) as expected from laboratory measurements are presented. Experimental results at energies over 1 MeV are in very good agreement with our calculations for pure nitrogen while discrepancies of about 20% are found for dry air, very likely associated to uncertainties in the available data on quenching cross sections. The relationship between fluorescence emission, stopping power and deposited energy is discussed.

  3. Application of magnetic and core-shell nanoparticles to determine enrofloxacin and its metabolite using laser induced fluorescence microscope.

    Science.gov (United States)

    Kim, Suji; Ko, Junga; Lim, H B

    2013-04-10

    A unique analytical method using nanoparticles and laser-induced fluorescence microscopy (LIFM) was developed to determine enrofloxacin in this work. For sample pretreatment, two different kinds of particles, i.e., synthesized dye-doped core-shell silica nanoparticles and magnetic micro-particles (MPs), were used for fluorescent tagging and concentrating the enrofloxacin, respectively. The antibody of enrofloxacin was immobilized on the synthesized FITC-doped core-shell nanoparticles, and the enrofloxacin target was extracted by the MPs. At this moment, the average number of antibodies on each core-shell silica nanoparticle was ~0.9, which was determined by the fluorescence ratiometric method. The described method was demonstrated for a meat sample to determine enrofloxacin using LIFM, and the result was compared with enzyme-linked immunosorbent assay (ELISA). The developed technique allowed the simplified analytical procedure, improved the detection limit about 54-fold compared to ELISA.

  4. Quantification of alginate by aggregation induced by calcium ions and fluorescent polycations.

    Science.gov (United States)

    Zheng, Hewen; Korendovych, Ivan V; Luk, Yan-Yeung

    2016-01-01

    For quantification of polysaccharides, including heparins and alginates, the commonly used carbazole assay involves hydrolysis of the polysaccharide to form a mixture of UV-active dye conjugate products. Here, we describe two efficient detection and quantification methods that make use of the negative charges of the alginate polymer and do not involve degradation of the targeted polysaccharide. The first method utilizes calcium ions to induce formation of hydrogel-like aggregates with alginate polymer; the aggregates can be quantified readily by staining with a crystal violet dye. This method does not require purification of alginate from the culture medium and can measure the large amount of alginate that is produced by a mucoid Pseudomonas aeruginosa culture. The second method employs polycations tethering a fluorescent dye to form suspension aggregates with the alginate polyanion. Encasing the fluorescent dye in the aggregates provides an increased scattering intensity with a sensitivity comparable to that of the conventional carbazole assay. Both approaches provide efficient methods for monitoring alginate production by mucoid P. aeruginosa.

  5. Determination of phenylurea pesticides by direct laser photo-induced fluorescence.

    Science.gov (United States)

    Diaw, P A; Maroto, A; Mbaye, O M A; Gaye-Seye, M D; Stephan, L; Coly, A; Deschamps, L; Tine, A; Aaron, J J; Giamarchi, P

    2013-11-15

    A direct Laser Photo-Induced Fluorescence (DL-PIF) method is developed for the determination of two phenylurea pesticides, namely fenuron and diflubenzuron. The DL-PIF method uses a tunable Nd:YAG-OPO Laser to obtain the photoproduct(s) and to simultaneously analyse their fluorescence in a short acquisition time on an intensified CCD camera. Compared to classical PIF methods, the use of a tunable laser improves the selectivity (by choosing the suitable excitation wavelength), increases the sensitivity (due to the high energy of the beam) and also reduces the time of analysis. The analytical performances of this method for the determination of both pesticides are satisfactory in comparison to other classical PIF methods published for the determination of phenylurea pesticides. The calibration curves were linear over one order of magnitude and the limits of detection were in the ng mL(-1) range. Satisfactory recoveries were obtained in the analysis of both pesticides in river and sea water spiked samples. Copyright © 2013 Elsevier B.V. All rights reserved.

  6. Use of a laser-induced fluorescence thermal imaging system for film cooling heat transfer measurement

    Energy Technology Data Exchange (ETDEWEB)

    Chyu, M.K. [Carnegie Mellon Univ., Pittsburgh, PA (United States)

    1995-10-01

    This paper describes a novel approach based on fluorescence imaging of thermographic phosphor that enables the simultaneous determination of both local film effectiveness and local heat transfer on a film-cooled surface. The film cooling model demonstrated consists of a single row of three discrete holes on a flat plate. The transient temperature measurement relies on the temperature-sensitive fluorescent properties of europium-doped lanthanum oxysulfide (La{sub 2}O{sub 2}S:EU{sup 3+}) thermographic phosphor. A series of full-field surface temperatures, mainstream temperatures, and coolant film temperatures were acquired during the heating of a test surface. These temperatures are used to calculate the heat transfer coefficients and the film effectiveness simultaneously. Because of the superior spatial resolution capability for the heat transfer data reduced from these temperature frames, the laser-induced fluorescence (LIF) imaging system, the present study observes the detailed heat transfer characteristics over a film-protected surface. The trend of the results agrees with those obtained using other conventional thermal methods, as well as the liquid crystal imaging technique. One major advantage of this technique is the capability to record a large number of temperature frames over a given testing period. This offers multiple-sample consistency.

  7. Determination of imidacloprid in water samples via photochemically induced fluorescence and second-order multivariate calibration.

    Science.gov (United States)

    Fuentes, Edwar; Cid, Camila; Báez, María E

    2015-03-01

    This paper presents a new method for the determination of imidacloprid in water samples; one of the most widely used neonicotinoid pesticides in the farming industry. The method is based on the measurement of excitation-emission spectra of photo-induced fluorescence (PIF-EEMs) associated with second-order multivariate calibration with a parallel factor analysis (PARAFAC) and unfolded partial least squares coupled to residual bilinearization (U-PLS/RBL). The second order advantage permitted the determination of imidacloprid in the presence of potential interferences, which also shows photo-induced fluorescence (other pesticides and/or unexpected compounds of the real samples). The photoreaction was performed in 100-μl disposable micropipettes. As a preliminary step, solid phase extraction on C18 (SPE-C18) was applied to concentrate the analyte and diminish the limit of detection. The LOD was approximately 1 ng mL(-1), which is suitable for detecting imidacloprid in water according to the guidelines established in North America and Europe. The PIF-EEMs coupled to PARAFAC or U-PLS/RBL was successfully applied for the determination of imidacloprid in different real water samples, with an average recovery of 101±10%.

  8. Determination of RNA degradation by capillary electrophoresis with cyan light-emitted diode-induced fluorescence.

    Science.gov (United States)

    Yang, Tzu-Hsueh; Chang, Po-Ling

    2012-05-25

    RNA integrity plays an important role in RNA studies because poor RNA quality may have a great impact on downstream methodologies. This study proposes a cost-effective, rapid, and sensitive method for determining RNA integrity based on capillary electrophoresis that utilizes a cyan light-emitted diode-induced fluorescence as a separation tool. The capillary was initially coated with 0.1% Poly(vinylpyrrolidone) (M(ave) 1,300,000 Da) to reduce electroosmotic flow and avoid RNA adsorption. When the capillary was filled with 0.4% poly(ethylene) oxide (M(ave) 4,000,000) and a nucleic acid-specific fluorescent dye, SYTO 9, the baseline separation of the 18S and 28S ribosomal RNAs (rRNAs) in total RNA was accomplished within 15 min. The lowest detectable concentration for the 18S and 28S rRNAs was estimated to be 50 pg/μL. Some peaks longer than the 28S rRNA that migrated slowly were observed as long as the initial total RNA concentration was optimized. The temperature-induced degradation of the large RNA fragments (longer than the 28S rRNA) was faster than that of 18S rRNA and 28S rRNA. These large RNA fragments may serve as a promising marker for testing RNA integrity compared to the traditional method. Copyright © 2012 Elsevier B.V. All rights reserved.

  9. UV laser-induced fluorescence spectroscopy and laser Doppler flowmetry in the diagnostics of alopecia

    Science.gov (United States)

    Skomorokha, Diana P.; Pigoreva, Yulia N.; Salmin, Vladimir V.

    2016-04-01

    Development of optical biopsy methods has a great interest for medical diagnostics. In clinical and experimental studies it is very important to analyze blood circulation quickly and accurately, thereby laser Doppler flowmetry (LDF) is widely used. UV laser-induced fluorescence spectroscopy (UV LIFS) is express highly sensitive and widely-spread method with no destructive impact, high excitation selectivity and the possibility to use in highly scattering media. The goal of this work was to assess a correlation of UV laser-induced fluorescence spectroscopy and laser Doppler flowmetry parameters, and a possibility to identify or to differentiate various types of pathological changes in tissues according to their autofluorescence spectra. Three groups of patients with diffuse (symptomatic) alopecia, androgenic alopecia, and focal alopecia have been tested. Each groups consisted of not less than 20 persons. The measurements have been done in the parietal and occipital regions of the sculls. We used the original automated spectrofluorimeter to record autofluorescence spectra, and standard laser Doppler flowmeter BLF-21 (Transonic Systems, Inc., USA) to analyze the basal levels of blood circulation. Our results show that UV LIFS accurately distinguishes the zones with different types of alopecia. We found high correlation of the basal levels of blood circulation and the integrated intensity of autofluorescence in the affected tissue.

  10. Constitutive and Inducible Expression of Green Fluorescent Protein in Brucella suis

    Science.gov (United States)

    Köhler, Stephan; Ouahrani-Bettache, Safia; Layssac, Marion; Teyssier, Jacques; Liautard, Jean-Pierre

    1999-01-01

    A gene fusion system based on plasmid pBBR1MCS and the expression of green fluorescent protein was developed for Brucella suis, allowing isolation of constitutive and inducible genes. Bacteria containing promoter fusions of chromosomal DNA to gfp were visualized by fluorescence microscopy and examined by flow cytometry. Twelve clones containing gene fragments induced inside J774 murine macrophages were isolated and further characterized. PMID:10569794

  11. Constitutive and Inducible Expression of Green Fluorescent Protein in Brucella suis

    OpenAIRE

    Köhler, Stephan; Ouahrani-Bettache, Safia; Layssac, Marion; Teyssier, Jacques; Liautard, Jean-Pierre

    1999-01-01

    A gene fusion system based on plasmid pBBR1MCS and the expression of green fluorescent protein was developed for Brucella suis, allowing isolation of constitutive and inducible genes. Bacteria containing promoter fusions of chromosomal DNA to gfp were visualized by fluorescence microscopy and examined by flow cytometry. Twelve clones containing gene fragments induced inside J774 murine macrophages were isolated and further characterized.

  12. Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR)

    DEFF Research Database (Denmark)

    Skeldal, Sune; Kjaergaard, Maj M; Alwasel, Saleh

    2015-01-01

    the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay...... to monitor the interaction between fluorescently tagged sortilin and p75(NTR) in live cells. The method is based on a standard fluorescent plate reader found in many biochemical laboratories and the results are evaluated using a microscopy-based quantified sensitized acceptor emission FRET approach making...

  13. Understanding Solar Induced Fluorescence: Building up from Leaf Scale Measurements (Invited)

    Science.gov (United States)

    Berry, J. A.; Van der Tol, C.; Frankenberg, C.; Joiner, J.; Guanter, L.

    2013-12-01

    Measurements of chlorophyll fluorescence have long been a key method for probing the mechanisms of photosynthesis in laboratory studies. Recent advances in satellite spectroscopy have enabled retrieval of chlorophyll fluorescence from terrestrial ecosystems at a global scale. Analyses of these retrievals show promising potential as an indicator of photosynthetic rate and of its response to environmental stress. This talk will explore the mechanistic basis for interpreting and modeling of solar induced chlorophyll fluorescence ( SIF). SIF is essentially a leak of photons from photosynthetic membranes, and it is, therefore, related to the flux of photons absorbed by chlorophyll and to biochemical processes that regulate the processing of these photons in macromolecuar complexes associated with photosystem II. Thus: SIF = aPAR * φF, where aPAR is the flux of absorbed photosynthetically active radiation and φF, is the yield (light-use efficiency) of fluorescence. (For simplicity we will ignore the transport of fluorescence from its sources to the sensor for the moment). This expression for SIF is similar to a common expression for photosynthesis or gross primary productivity, GPP = aPAR * LUE, where LUE, is the light-use-efficiency for CO2 uptake. These equations can be combined and simplified to illustrate the relationship between SIF and GPP; GPP =SIF *LUE / φF. The extent to which GPP is proportional to SIF hinges on the stability of the ratio, LUE / φF, and it leads to the key question to be considered here. What is the relationship between the light-use-efficiency for photosynthesis and that for fluorescence? Satellite retrievals of SIF occur at mid-day, conditions where the capacity for CO2 fixation usually limits the rate of photosynthesis. Under this condition the rate of the photo-acts must be down-regulated to protect from photo-damage. This balancing the source with the sink is accomplished by opening non-photochemical trapping centers that compete with

  14. Determination of metoprolol in rabbit blood using capillary electrophoresis with laser-induced fluorescence detection

    Institute of Scientific and Technical Information of China (English)

    Yu Yun Chen; Wei Ping Yang; Zhu Jun Zhang

    2011-01-01

    This work described a sensitive method for determination of metoprolol in rabbit plasma. The method involved purification by ultrafiltration, derivatization with fluorescein isothiocyanate, determination by capillary electrophoresis (CE) coupled with laser-induced fluorescence (LIF) detector. Other components in plasma including a variety of amino acids and proteins did not interfere with the determination of metoprolol in experimental condition. The assay had a wide range (2.0-500 ng/mL) of linearity and a detection limit of 0.8 ng/mL. The intra-and inter-day precisions were satisfactory with relative standard deviation (RSD) less than 10.0% and accuracy within 10.0%. This method was successfully applied to pharmacokinetic study of metoprolol in rabbit blood. (c) 2010 Yu Yun Chen. Published by Elsevier B.V. on behalf of Chinese Chemical Society. All rights reserved.

  15. Flow injection fluorescence determination of dopamine using a photo induced electron transfer (PET) boronic acid derivative

    Energy Technology Data Exchange (ETDEWEB)

    Ebru Seckin, Z. [Department of Chemistry, Middle East Technical University, 06531 Ankara (Turkey); Volkan, Muervet [Department of Chemistry, Middle East Technical University, 06531 Ankara (Turkey)]. E-mail: murvet@metu.edu.tr

    2005-08-15

    An automated flow injection analysis system was developed for the fluorometric determination of dopamine in pharmaceutical injections. The method is based on the quenching effect of dopamine on m-dansylaminophenyl boronic acid (DAPB) fluorescence due to the reverse photo induced electron transfer (PET) mechanism. Effects of pH and interfering species on the determination of dopamine were examined. Calibration for dopamine, based on quenching data, was linear in the concentration range of 1.0 x 10{sup -5} to 1.0 x 10{sup -4} M. Detection limit (3 s) of the method was found to be 3.7 x 10{sup -6} M. Relative standard deviation of 1.2% (n = 10) was obtained with 1.0 x 10{sup -5} M dopamine standard solution. The proposed method was applied successfully for the determination of dopamine in pharmaceutical injection sample. The sampling rate was determined as 24 samples per hour.

  16. The application of time decay characteristics of laser-induced fluorescence in the classification of vegetation.

    Science.gov (United States)

    Gong, Wei; Yang, Jian; Shi, Shuo; Du, Lin; Sun, Jia; Song, Shalei

    2017-02-01

    In this study, the time decay of the chlorophyll fluorescence intensity (TDCFI) of vegetation was measured based on laser-induced fluorescence (LIF) technology with a 355 nm laser serving as the excitation light source. The pseudo-color diagram of the TDCFI (PDTDCFIs) was proposed for use as a characteristic fingerprint for the analysis of various plant species based on variations in the fluorescence intensity over time. Compared with the steady-state fluorescence spectra, two-dimensional PDTDCFIs contained more spectral information, including variations in both the shape of the laser-induced fluorescence spectra and the relative intensity. The experimental results demonstrated that the PDTDCFIs of various plant species show distinct differences, and this was successfully applied in the classification of plant species. Therefore, the PDTDCFIs of plants could provide researchers with a more reliable and useful tool for the characterization of vegetation. Copyright © 2016 John Wiley & Sons, Ltd. Copyright © 2016 John Wiley & Sons, Ltd.

  17. Development of time-resolved laser-induced fluorescence spectroscopic technique for the analysis of biomolecules

    Directory of Open Access Journals (Sweden)

    Terzić M.

    2008-01-01

    Full Text Available Our developments of the time-resolved laser-induced fluorescence (TR-LIF detection system for biomolecules are presented. This system is based on the tunable (320 nm to 475 nm Nd:YAG laser pulses used to excite various biomolecules. The detection part is the Streak System for Fluorescence Lifetime Spectroscopy (Hamamatsu, Japan. The system consists of a C4334-01 streakscope, as a detector, DG 535 digital pulse/delay generator, C5094-S Spectrograph and HPD-TA System, as a temporal analyzer. The TR-LIF spectrometer is designed primarily to study the temperature and pressure effects on fluorescence behavior of biomolecules upon excitation with a single nanosecond pulse. The design of this system has capability to combine laser-induced breakdown (LIB with fluorescence, as well to study optodynamic behavior of fluorescence biomolecules.

  18. [Laser induced fluorescence spectrum characteristics of common edible oil and fried cooking oil].

    Science.gov (United States)

    Mu, Tao-tao; Chen, Si-ying; Zhang, Yin-chao; Chen, He; Guo, Pan; Ge, Xian-ying; Gao, Li-lei

    2013-09-01

    In order to detect the trench oil the authors built a trench oil rapid detection system based on laser induced fluorescence detection technology. This system used 355 nm laser as excitation light source. The authors collected the fluorescence spectrum of a variety of edible oil and fried cooking oil (a kind of trench oil) and then set up a fluorescence spectrum database by taking advantage of the trench oil detection system It was found that the fluorescence characteristics of fried cooking oil and common edible oil were obviously different. Then it could easily realize the oil recognition and trench oil rapid detection by using principal component analysis and BP neural network, and the overall recognition rate could reach as high as 97.5%. Experiments showed that laser induced fluorescence spectrum technology was fast, non-contact, and highly sensitive. Combined with BP neural network, it would become a new technique to detect the trench oil.

  19. Teaching laser-induced fluorescence of plant leaves

    Science.gov (United States)

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

    2016-11-01

    Plants convert carbon dioxide into sugars using the energy of sunlight. Absorbed light unused for conversion is dissipated primarily as heat with a small fraction re-emitted as fluorescence at longer wavelengths. One can use the latter to estimate photosynthetic activity. The illumination of intact leaves with strong light after keeping them in dark for tens of minutes results in a rapid increase followed by a slow decay of fluorescence emission from the fluorophore chlorophyll-a, called the Kautsky effect. This paper describes a laboratory practice that introduces students of physics or engineering into this research field. It begins with the spectral measurement of the fluorescence emitted by a plant leaf upon UV excitation. Then it focuses on the red and far-red components of the fluorescence emission spectrum characteristic to the chlorophyll-a molecule and presents an inexpensive demonstration of the Kautsky effect. As researchers use more complex measurement techniques and tools, the practice ends up with the demonstration of an intelligent fluorosensor, a compact tool developed for plant physiological research and horticulture applications together with a brief interpretation of some important fluorescence parameters.

  20. Early-photon guided reconstruction method for time-domain fluorescence lifetime tomography

    Institute of Scientific and Technical Information of China (English)

    Lin Zhang; Chuangjian Cai; Yanlu Lv; Jianwen Luo

    2016-01-01

    A reconstruction method guided by early-photon fluorescence yield tomography is proposed for time-domain fluorescence lifetime tomography (FLT) in this study.The method employs the early-arriving photons to reconstruct a fluorescence yield map,which is utilized as a priori information to reconstruct the FLT via all the photons along the temporal-point spread functions.Phantom experiments demonstrate that,compared with the method using all the photons for reconstruction of fluorescence yield and lifetime maps,the proposed method can achieve higher spatial resolution and reduced crosstalk between different targets without sacrificing the quantification accuracy of lifetime and contrast between heterogeneous targets.

  1. Evaluation of path-history-based fluorescence Monte Carlo method for photon migration in heterogeneous media.

    Science.gov (United States)

    Jiang, Xu; Deng, Yong; Luo, Zhaoyang; Wang, Kan; Lian, Lichao; Yang, Xiaoquan; Meglinski, Igor; Luo, Qingming

    2014-12-29

    The path-history-based fluorescence Monte Carlo method used for fluorescence tomography imaging reconstruction has attracted increasing attention. In this paper, we first validate the standard fluorescence Monte Carlo (sfMC) method by experimenting with a cylindrical phantom. Then, we describe a path-history-based decoupled fluorescence Monte Carlo (dfMC) method, analyze different perturbation fluorescence Monte Carlo (pfMC) methods, and compare the calculation accuracy and computational efficiency of the dfMC and pfMC methods using the sfMC method as a reference. The results show that the dfMC method is more accurate and efficient than the pfMC method in heterogeneous medium.

  2. A new relative referencing method for crop monitoring using chlorophyll fluorescence

    Science.gov (United States)

    Norikane, J.; Goto, E.; Kurata, K.; Takakura, T.

    The measurement of plant chlorophyll fluorescence has been used for many years as a method to monitor a plant's health status. These types of methods have been mostly relegated to the laboratory. The newly developed Relative Referencing Method allows for the measurement of chlorophyll fluorescence under artificial lighting conditions. The fluorescence signal can be determined by first taking a reference signal measurement, then a second measurement with an additional fluorescence excitation source. The first signal can then be subtracted from the second and the plant's chlorophyll fluorescence due to the second lighting source can be determined. With this simple approach, a photosynthesizing plant can be monitored to detect signs of water stress. Using this approach experiments on tomato plants have shown that it was possible to detect water stress, while the plants were continuously illuminated by fluorescent lamps. This method is a promising tool for the remote monitoring of crops grown in a CELSS-type application.

  3. Variability of the specific fluorescence of chlorophyll in the ocean. Part 2. Fluorometric method of chlorophyll a determination

    Directory of Open Access Journals (Sweden)

    Miros³awa Ostrowska

    2000-06-01

    Full Text Available Two methods of determining the chlorophyll a concentration in the sea have been formulated on the basis of artificially induced fluorescence measured with the aid of submersible fluorometers. The method of statistical correlation is founded on the empirical relationship between fluorescence and chlorophyll concentration. The theoretical model of fluorescence described in Part 1 of this paper (see Ostrowska et al. 2000, this volume provides the basis of the other method, the physical method. This describes the dependence of the specific fluorescence of phytoplankton on the chlorophyll concentration, a diversity of photophysiological properties of phytoplankton and the optical characteristics of the fluorometer.     In order to assess their practicability, the methods were subjected to empirical verification. This showed that the physical method yielded chlorophyll concentrations of far greater accuracy. The respective error factors of the estimated chlorophyll concentration were x = 2.07 for the correlation method and x = 1.5 for the physical method. This means that the statistical logarithmic error varies from -52 to +107% in the case of the former method but only from -33 to +51% in the case of the latter. Thus, modifying the methodology has much improved the accuracy of chlorophyll determinations.

  4. Investigation on the photo-induced de-oxygenation process of myoglobin in aqueous solution by use of fluorescence spectroscopy

    Institute of Scientific and Technical Information of China (English)

    2008-01-01

    A photo-induced de-oxygenation process of myoglobin (Mb) in aqueous solution was investigated by use of fluorescence spectroscopy. The spectra are characterized by the fluorescence intensity declining gradually after each scan,and the decay of fluorescence intensity being significant in each scan,which is assigned to the release of oxygen from the opening of the heme-pockets induced by illumination. More illumination will cause more release of oxygen; if the temperature of an Mb solution is increased when it is illuminated,the rate of de-oxygenation will be higher. It was found that ligand-oxygen in Fe-porphyrin could be removed from Mb by nitrogen. This indicates that the interac-tion between oxy-Mb and other different gases can be tested by the method of fluorescence spectros-copy. In addition,fluorescence spectroscopy can be employed to probe the energy transfer between Fe-porphyrin and tryptophan or tyrosine in Mb molecules.

  5. Fundamentals and practice for ultrasensitive laser-induced fluorescence detection in microanalytical systems.

    Science.gov (United States)

    Johnson, Mitchell E; Landers, James P

    2004-11-01

    Laser-induced fluorescence is an extremely sensitive method for detection in chemical separations. In addition, it is well-suited to detection in small volumes, and as such is widely used for capillary electrophoresis and microchip-based separations. This review explores the detailed instrumental conditions required for sub-zeptomole, sub-picomolar detection limits. The key to achieving the best sensitivity is to use an excitation and emission volume that is matched to the separation system and that, simultaneously, will keep scattering and luminescence background to a minimum. We discuss how this is accomplished with confocal detection, 90 degrees on-capillary detection, and sheath-flow detection. It is shown that each of these methods have their advantages and disadvantages, but that all can be used to produce extremely sensitive detectors for capillary- or microchip-based separations. Analysis of these capabilities allows prediction of the optimal means of achieving ultrasensitive detection on microchips.

  6. Pyrene measurements in sooting low pressure methane flames by jet-cooled laser-induced fluorescence.

    Science.gov (United States)

    Wartel, M; Pauwels, J-F; Desgroux, P; Mercier, X

    2011-12-15

    This paper presents in detail the study we carried out concerning the pyrene measurement by jet-cooled laser-induced fluorescence (JCLIF) in different sooting low pressure methane flames. The aim of this paper is both to demonstrate the potentialities of this technique for the measurement of such moderately sized polycyclic aromatic hydrocarbons under sooting flame conditions and to provide new experimental data for the understanding and the development of chemical models of the soot formation processes. Several concentration profiles of pyrene measured in different sooting flame (various pressure and equivalence ratio) are presented. The validation of the JCLIF method for pyrene measurements is explained in detail as well as the calibration procedure, based on the standard addition method, which has been implemented for the quantification of the concentration profiles. Sensitivity lower than 1 ppb was obtained for the measurement of this species under sooting flame conditions.

  7. Improving the fluorescence polarization method to evaluate the orientation of fluorescent systems adsorbed in ordered layered materials

    Energy Technology Data Exchange (ETDEWEB)

    Salleres, Sandra [Departamento de Quimica Fisica, Universidad del Pais Vasco UPV/EHU, Apartado 644, 48080 Bilbao (Spain); Arbeloa, Fernando Lopez, E-mail: fernando.lopezarbeloa@ehu.e [Departamento de Quimica Fisica, Universidad del Pais Vasco UPV/EHU, Apartado 644, 48080 Bilbao (Spain); Martinez, Virginia Martinez; Arbeloa, Teresa; Lopez Arbeloa, Inigo [Departamento de Quimica Fisica, Universidad del Pais Vasco UPV/EHU, Apartado 644, 48080 Bilbao (Spain)

    2009-11-15

    The fluorescence polarization method, recently developed for the evaluation of the preferential orientation of fluorescent dyes adsorbed in layered materials [F. Lopez Arbeloa, V. Martinez Martinez, J. Photochem. Photobiol. A: Chem. 181 (2006) 44], is readapted to improve its application. Fluorescence polarization was previously obtained by recording the emission intensity for two orthogonal orientations of the emission polarizer (i.e., the horizontal and vertical polarized light) after excitation with vertical or horizontal polarized light. In the method proposed in this work, samples are excited with unpolarized light, reducing the polarization effect of the excitation light scattering at those emission wavelengths close to the excitation wavelength. Moreover, the present method decreases the effect of the orientation of other non-fluorescent species present in the system, which are active in the excitation process. Consequently, the new method is more simple, precise and sensitive. It is applied to evaluate the orientation of rhodamine 6G dye adsorbed in ordered laponite clay films with low and moderated dye loadings.

  8. Absolute absorption and fluorescence measurements over a dynamic range of 10$^6$ with cavity-enhanced laser-induced fluorescence

    CERN Document Server

    Sanders, Scott E; Nahler, N Hendrik; Wrede, Eckart

    2013-01-01

    We describe a novel experimental setup that combines the advantages of both laser-induced fluorescence and cavity ring-down techniques. The simultaneous measurement of the ring-down and fluorescence signals from the same sample in a single laser beam delivers the calibration of the fluorescence measurement to gain absolute quantities: absorption cross section, sample density and fluorescence quantum yield. At the same time, the fluorescence measurement extends the dynamic range of a stand-alone cavity ring-down setup from typically three to at least six orders of magnitude. The methodology of this combined cavity-enhanced laser-induced fluorescence (CELIF) technique is developed and rigorously tested against the spectroscopy of 1,4-bis(phenylethynyl)benzene in a molecular beam and density measurements in a cell.

  9. A new post-column reactor-laser induced fluorescence detector for capillary electrophoresis

    Energy Technology Data Exchange (ETDEWEB)

    Liling, Zhang [Iowa State Univ., Ames, IA (United States)

    1996-01-02

    Capillary zone electrophoresis (CZE), a powerful separation method based on the differential migration of charged species under the influence of an electric field, has been widely used for separations covering from small ions to big biomolecules. Chapter 1 describes the method, then discusses detection of the separated analytes by laser induced fluorescence and by chemical derivatization, and the use of O-phthaldialdehyde (OPA) as a post-column reagent. Chapter 2 describes a post-column reactor which uses two narrow bore capillaries connected coaxially. This reactor differs from other coaxial reactors in terms of capillary dimensions, reagent flow control, ease of construction and most importantly, better limits of detection. The derivatization reagent is electroosmotically driven into the reaction capillary and the reagent flow rate is independently controlled by a high voltage power supply. Amino acids, amines and proteins, derivatized by OPA/2-mercaptoethanol using this post-column reactor coupled with LIF detection, show low attomole mass limits of detection, and for the first time, the authors demonstrate single cell capability with a post-column derivatization scheme. The single cell capability shows that this reactor could find applications in assaying non-fluorescent or electrochemically inactive components in individual biological cells in the future.

  10. In vitro quantitative light-induced fluorescence to measure changes in enamel mineralization.

    Science.gov (United States)

    Gmür, Rudolf; Giertsen, Elin; van der Veen, Monique H; de Josselin de Jong, Elbert; ten Cate, Jacob M; Guggenheim, Bernhard

    2006-09-01

    A sensitive, quantitative method for investigating changes in enamel mineralization of specimens subjected to in vitro or in situ experimentation is presented. The fluorescence-detecting instrument integrates a Xenon arc light source and an object positioning stage, which makes it particularly suitable for the nondestructive assessment of demineralized or remineralized enamel. We demonstrate the ability of in vitro quantitative light-induced fluorescence (QLF) to quantify changes in mineralization of bovine enamel discs that had been exposed in vitro to a demineralizing gel (n=36) or biofilm-mediated demineralization challenges (n=10), or were carried in situ by three volunteers during a 10-day experiment (n=12). Further experiments show the technique's value for monitoring the extent of remineralization in 36 specimens exposed in vitro to oral multispecies biofilms and document the repeatability of in vitro QLF measurements (n=10) under standardized assay conditions. The validity of the method is illustrated by comparison with transversal microradiography (TMR), the invasive current gold standard for assessing experimental changes in enamel mineralization. Ten discs with 22 measurement areas for comparison demonstrated a positive correlation between TMR and QLF (r=0.82). Filling a technological gap, this QLF system is a promising tool to assay in vitro nondestructively localized changes in mineralization of enamel specimens.

  11. Stereoscopic Imaging in Hypersonics Boundary Layers using Planar Laser-Induced Fluorescence

    Science.gov (United States)

    Danehy, Paul M.; Bathel, Brett; Inman, Jennifer A.; Alderfer, David W.; Jones, Stephen B.

    2008-01-01

    Stereoscopic time-resolved visualization of three-dimensional structures in a hypersonic flow has been performed for the first time. Nitric Oxide (NO) was seeded into hypersonic boundary layer flows that were designed to transition from laminar to turbulent. A thick laser sheet illuminated and excited the NO, causing spatially-varying fluorescence. Two cameras in a stereoscopic configuration were used to image the fluorescence. The images were processed in a computer visualization environment to provide stereoscopic image pairs. Two methods were used to display these image pairs: a cross-eyed viewing method which can be viewed by naked eyes, and red/blue anaglyphs, which require viewing through red/blue glasses. The images visualized three-dimensional information that would be lost if conventional planar laser-induced fluorescence imaging had been used. Two model configurations were studied in NASA Langley Research Center's 31-Inch Mach 10 Air Wind tunnel. One model was a 10 degree half-angle wedge containing a small protuberance to force the flow to transition. The other model was a 1/3-scale, truncated Hyper-X forebody model with blowing through a series of holes to force the boundary layer flow to transition to turbulence. In the former case, low flowrates of pure NO seeded and marked the boundary layer fluid. In the latter, a trace concentration of NO was seeded into the injected N2 gas. The three-dimensional visualizations have an effective time resolution of about 500 ns, which is fast enough to freeze this hypersonic flow. The 512x512 resolution of the resulting images is much higher than high-speed laser-sheet scanning systems with similar time response, which typically measure 10-20 planes.

  12. DETECTION OF LOW DOSE RADIATION INDUCED DNA DAMAGE USING TEMPERATURE DIFFERENTIAL FLUORESCENCE ASSAY

    Science.gov (United States)

    A rapid and sensitive fluorescence assay for radiation-induced DNA damage is reported. Changes in temperature-induced strand separation in both calf thymus DNA and plasmid DNA (puc 19 plasmid from Escherichia coli) were measured after exposure to low doses of radiation. Exposur...

  13. Development of a quantitative diagnostic method of estrogen receptor expression levels by immunohistochemistry using organic fluorescent material-assembled nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Gonda, Kohsuke, E-mail: gonda@med.tohoku.ac.jp [Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8575 (Japan); Miyashita, Minoru [Department of Surgical Oncology, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8574 (Japan); Watanabe, Mika; Takahashi, Yayoi [Department of Pathology, Tohoku University Hospital, Seiryo-machi, Aoba-ku, Sendai 980-8574 (Japan); Goda, Hideki; Okada, Hisatake; Nakano, Yasushi [Optical and Biological R and D Center, Konica Minolta Technology Center, Inc., No. 1 Sakuramachi, Hino-shi, Tokyo 191-8511 (Japan); Tada, Hiroshi [Department of Surgical Oncology, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8574 (Japan); Amari, Masakazu [Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8575 (Japan); Ohuchi, Noriaki [Department of Nano-Medical Science, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8575 (Japan); Department of Surgical Oncology, Graduate School of Medicine, Tohoku University, Seiryo-machi, Aoba-ku, Sendai 980-8574 (Japan)

    2012-09-28

    Highlights: Black-Right-Pointing-Pointer Organic fluorescent material-assembled nanoparticles for IHC were prepared. Black-Right-Pointing-Pointer New nanoparticle fluorescent intensity was 10.2-fold greater than Qdot655. Black-Right-Pointing-Pointer Nanoparticle staining analyzed a wide range of ER expression levels in tissue. Black-Right-Pointing-Pointer Nanoparticle staining enhanced the quantitative sensitivity for ER diagnosis. -- Abstract: The detection of estrogen receptors (ERs) by immunohistochemistry (IHC) using 3,3 Prime -diaminobenzidine (DAB) is slightly weak as a prognostic marker, but it is essential to the application of endocrine therapy, such as antiestrogen tamoxifen-based therapy. IHC using DAB is a poor quantitative method because horseradish peroxidase (HRP) activity depends on reaction time, temperature and substrate concentration. However, IHC using fluorescent material provides an effective method to quantitatively use IHC because the signal intensity is proportional to the intensity of the photon excitation energy. However, the high level of autofluorescence has impeded the development of quantitative IHC using fluorescence. We developed organic fluorescent material (tetramethylrhodamine)-assembled nanoparticles for IHC. Tissue autofluorescence is comparable to the fluorescence intensity of quantum dots, which are the most representative fluorescent nanoparticles. The fluorescent intensity of our novel nanoparticles was 10.2-fold greater than quantum dots, and they did not bind non-specifically to breast cancer tissues due to the polyethylene glycol chain that coated their surfaces. Therefore, the fluorescent intensity of our nanoparticles significantly exceeded autofluorescence, which produced a significantly higher signal-to-noise ratio on IHC-imaged cancer tissues than previous methods. Moreover, immunostaining data from our nanoparticle fluorescent IHC and IHC with DAB were compared in the same region of adjacent tissues sections to

  14. Evaluation of dental enamel caries assessment using Quantitative Light Induced Fluorescence and Optical Coherence Tomography.

    Science.gov (United States)

    Maia, Ana Marly Araújo; de Freitas, Anderson Zanardi; de L Campello, Sergio; Gomes, Anderson Stevens Leônidas; Karlsson, Lena

    2016-06-01

    An in vitro study of morphological alterations between sound dental structure and artificially induced white spot lesions in human teeth, was performed through the loss of fluorescence by Quantitative Light-Induced Fluorescence (QLF) and the alterations of the light attenuation coefficient by Optical Coherence Tomography (OCT). To analyze the OCT images using a commercially available system, a special algorithm was applied, whereas the QLF images were analyzed using the software available in the commercial system employed. When analyzing the sound region against white spot lesions region by QLF, a reduction in the fluorescence intensity was observed, whilst an increase of light attenuation by the OCT system occurred. Comparison of the percentage of alteration between optical properties of sound and artificial enamel caries regions showed that OCT processed images through the attenuation of light enhanced the tooth optical alterations more than fluorescence detected by QLF System. QLF versus OCT imaging of enamel caries: a photonics assessment.

  15. Strain Release Induced Novel Fluorescence Variation in CVD-Grown Monolayer WS2 Crystals.

    Science.gov (United States)

    Feng, Shanghuai; Yang, Ruilong; Jia, Zhiyan; Xiang, Jianyong; Wen, Fusheng; Mu, Congpu; Nie, Anmin; Zhao, Zhisheng; Xu, Bo; Tao, Chenggang; Tian, Yongjun; Liu, Zhongyuan

    2017-10-04

    Tensile strain is intrinsic to monolayer crystals of transition metal disulfides such as Mo(W)S2 grown on oxidized silicon substrates by chemical vapor deposition (CVD) owing to the much larger thermal expansion coefficient of Mo(W)S2 than that of silica. Here we report fascinating fluorescent variation in intensity with aging time in CVD-grown triangular monolayer WS2 crystals on SiO2 (300 nm)/Si substrates and formation of interesting concentric triangular fluorescence patterns in monolayer crystals of large size. The novel fluorescence aging behavior is recognized to be induced by the partial release of intrinsic tensile strain after CVD growth and the induced localized variations or gradients of strain in the monolayer crystals. The results demonstrate that strain has a dramatic impact on the fluorescence and photoluminescence of monolayer WS2 crystals and thus could potentially be utilized to tune electronic and optoelectronic properties of monolayer transition metal disulfides.

  16. Application of HPLC Combined with Laser Induced Fluorescence for Protein Profile Analysis of Tissue Homogenates in Cervical Cancer

    OpenAIRE

    Sujatha Bhat; Ajeetkumar Patil; Lavanya Rai; Kartha, V. B.; Santhosh Chidangil

    2012-01-01

    A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF) technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA) to get statistical information on different t...

  17. Direct probing of chromatography columns by laser-induced fluorescence. Technical progress report, September 1, 1989--February 28, 1993

    Energy Technology Data Exchange (ETDEWEB)

    McGuffin, V.L.

    1992-12-07

    This report summarizes the progress and accomplishments of this research project from September 1, 1989 to February 28, 1993. During this period, we have accomplished all of the primary scientific objectives of the research proposal: (1) constructed and evaluated a laser-induced fluorescence detection system that allows direct examination of the chromatographic column, (2) examined nonequilibrium processes that occur upon solute injection and elution, (3) examined solute retention in liquid chromatography as a function of temperature and pressure, (4) examined solute zone dispersion in liquid chromatography as a function of temperature and pressure, and (5) developed appropriate theoretical models to describe these phenomena. In each of these studies, substantial knowledge has been gained of the fundamental processes that are responsible for chromatographic separations. In addition to these primary research objectives, we have made significant progress in three related areas: (1) examined pyrene as a fluorescent polarity probe insupercritical fluids and liquids as a function of temperature and pressure, (2) developed methods for the class-selective identification of polynuclear aromatic hydrocarbons in coal-derived fluids by microcolumn liquid chromatography with fluorescence quenching detection, and (3) developed methods for the determination of saturated and unsaturated (including omega-3) fatty acids in fish oil extracts by microcolumn liquid chromatography with laser-induced fluorescence detection. In these studies, the advanced separation and detection techniques developed in our laboratory are applied to practical problems of environmental and biomedical significance.

  18. Analysis of laser-induced fluorescence spectra of in vitro plant tissue cultures

    Science.gov (United States)

    Muñoz-Muñoz, Ana Celia; Gutiérrez-Pulido, Humberto; Rodríguez-Domínguez, José Manuel; Gutiérrez-Mora, Antonia; Rodríguez-Garay, Benjamín; Cervantes-Martínez, Jesús

    2007-04-01

    We demonstrate the effectiveness of laser-induced fluorescence (LIF) for monitoring the development and stress detection of in vitro tissue cultures in a nondestructive and noninvasive way. The changes in LIF spectra caused by the induction of organogenesis, the increase of the F690/F740 ratio as a result of the stress originated in the organogenic explants due to shoot emergence, and the relationship between fluorescence spectra and shoot development were detected by LIF through closed containers of Saintpaulia ionantha.

  19. Indirect determination of the electric field in plasma discharges using laser-induced fluorescence spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Vaudolon, J., E-mail: julien.vaudolon@cnrs-orleans.fr; Mazouffre, S., E-mail: stephane.mazouffre@cnrs-orleans.fr [CNRS - ICARE (Institut de Combustion Aérothermique Réactivité et Environnement), 1 C Av. de la Recherche Scientifique, 45071 Orléans Cedex 2 (France)

    2014-09-15

    The evaluation of electric fields is of prime interest for the description of plasma characteristics. In this work, different methods for determining the electric field profile in low-pressure discharges using one- and two-dimensional Laser-Induced Fluorescence (LIF) measurements are presented and discussed. The energy conservation, fluid, and kinetic approaches appear to be well-suited for the electric field evaluation in this region of the plasma flow. However, the numerical complexity of a two-dimensional kinetic model is penalizing due to the limited signal-to-noise ratio that can be achieved, making the computation of the electric field subject to large error bars. The ionization contribution which appears in the fluid model makes it unattractive on an experimental viewpoint. The energy conservation and 1D1V kinetic approaches should therefore be preferred for the determination of the electric field when LIF data are used.

  20. Quantitative fluorescence using 5-aminolevulinic acid–induced protoporphyrin IX biomarker as a surgical adjunct in low-grade glioma surgery

    Science.gov (United States)

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

    2015-01-01

    OBJECT Previous studies in high-grade gliomas (HGGs) have indicated that protoporphyrin IX (PpIX) accumulates in higher concentrations in tumor tissue, and, when used to guide surgery, it has enabled improved resection leading to increased progression-free survival. Despite the benefits of complete resection and the advances in fluorescence-guided surgery, few studies have investigated the use of PpIX in low-grade gliomas (LGGs). Here, the authors describe their initial experience with 5-aminolevulinic acid (ALA)–induced PpIX fluorescence in a series of patients with LGG. METHODS Twelve patients with presumed LGGs underwent resection of their tumors after receiving 20 μg/kg of ALA approximately 3 hours prior to surgery under an institutional review board–approved protocol. Intraoperative assessments of the resulting PpIX emissions using both qualitative, visible fluorescence and quantitative measurements of PpIX concentration were obtained from tissue locations that were subsequently biopsied and evaluated histopathologically. Mixed models for random effects and receiver operating characteristic curve analysis for diagnostic performance were performed on the fluorescence data relative to the gold-standard histopathology. RESULTS Five of the 12 LGGs (1 ganglioglioma, 1 oligoastrocytoma, 1 pleomorphic xanthoastrocytoma, 1 oligodendroglioma, and 1 ependymoma) demonstrated at least 1 instance of visible fluorescence during surgery. Visible fluorescence evaluated on a specimen-by-specimen basis yielded a diagnostic accuracy of 38.0% (cutoff threshold: visible fluorescence score ≥ 1, area under the curve = 0.514). Quantitative fluorescence yielded a diagnostic accuracy of 67% (for a cutoff threshold of the concentration of PpIX [CPpIX] > 0.0056 μg/ml, area under the curve = 0.66). The authors found that 45% (9/20) of nonvisibly fluorescent tumor specimens, which would have otherwise gone undetected, accumulated diagnostically significant levels of CPpIX that were

  1. Single particle fluorescence burst analysis of epsin induced membrane fission.

    Science.gov (United States)

    Brooks, Arielle; Shoup, Daniel; Kustigian, Lauren; Puchalla, Jason; Carr, Chavela M; Rye, Hays S

    2015-01-01

    Vital cellular processes, from cell growth to synaptic transmission, rely on membrane-bounded carriers and vesicles to transport molecular cargo to and from specific intracellular compartments throughout the cell. Compartment-specific proteins are required for the final step, membrane fission, which releases the transport carrier from the intracellular compartment. The role of fission proteins, especially at intracellular locations and in non-neuronal cells, while informed by the dynamin-1 paradigm, remains to be resolved. In this study, we introduce a highly sensitive approach for the identification and analysis of membrane fission machinery, called burst analysis spectroscopy (BAS). BAS is a single particle, free-solution approach, well suited for quantitative measurements of membrane dynamics. Here, we use BAS to analyze membrane fission induced by the potent, fission-active ENTH domain of epsin. Using this method, we obtained temperature-dependent, time-resolved measurements of liposome size and concentration changes, even at sub-micromolar concentration of the epsin ENTH domain. We also uncovered, at 37°C, fission activity for the full-length epsin protein, supporting the argument that the membrane-fission activity observed with the ENTH domain represents a native function of the full-length epsin protein.

  2. An environmentally-friendly fluorescent method for quantification of lipid contents in yeast.

    Science.gov (United States)

    Poli, Jandora Severo; Lützhøft, Hans-Christian Holten; Karakashev, Dimitar Borisov; Valente, Patricia; Angelidaki, Irini

    2014-01-01

    This study aimed at developing an efficient, fast and environmentally-friendly method to quantify neutral lipid contents in yeast. After optimising the fluorescence instrument parameters and influence of organic solvent concentrations, a new method to quantify neutral lipids in yeast based on fluorescence was demonstrated. Isopropanol and Nile red in concentrations of 5% (final volume%) and 500 μg/L, respectively, were added to washed cells suspended in potassium chloride phosphate buffered saline (PBSKCl). Fluorescence was measured after 10 min in the dark. Glyceryltrioleate was used as model lipid and the calibration curve showed linearity (R(2)=0.994) between 0.50 and 25 mg/L. Compared with traditional gravimetric analysis, the developed method is much faster and uses less organic solvents. Lipid contents determined by fluorescence and gravimetry were the same for some strains, but for other strains the lipid contents determined by fluorescence were less. This new method will therefore be suitable for fast screening purposes.

  3. Investigation of Atmospheric Effects on Retrieval of Sun-Induced Fluorescence Using Hyperspectral Imagery

    Directory of Open Access Journals (Sweden)

    Zhuoya Ni

    2016-04-01

    Full Text Available Significant research progress has recently been made in estimating fluorescence in the oxygen absorption bands, however, quantitative retrieval of fluorescence data is still affected by factors such as atmospheric effects. In this paper, top-of-atmosphere (TOA radiance is generated by the MODTRAN 4 and SCOPE models. Based on simulated data, sensitivity analysis is conducted to assess the sensitivities of four indicators—depth_absorption_band, depth_nofs-depth_withfs, radiance and Fs/radiance—to atmospheric parameters (sun zenith angle (SZA, sensor height, elevation, visibility (VIS and water content in the oxygen absorption bands. The results indicate that the SZA and sensor height are the most sensitive parameters and that variations in these two parameters result in large variations calculated as the variation value/the base value in the oxygen absorption depth in the O2-A and O2-B bands (111.4% and 77.1% in the O2-A band; and 27.5% and 32.6% in the O2-B band, respectively. A comparison of fluorescence retrieval using three methods (Damm method, Braun method and DOAS and SCOPE Fs indicates that the Damm method yields good results and that atmospheric correction can improve the accuracy of fluorescence retrieval. Damm method is the improved 3FLD method but considering atmospheric effects. Finally, hyperspectral airborne images combined with other parameters (SZA, VIS and water content are exploited to estimate fluorescence using the Damm method and 3FLD method. The retrieval fluorescence is compared with the field measured fluorescence, yielding good results (R2 = 0.91 for Damm vs. SCOPE SIF; R2 = 0.65 for 3FLD vs. SCOPE SIF. Five types of vegetation, including ailanthus, elm, mountain peach, willow and Chinese ash, exhibit consistent associations between the retrieved fluorescence and field measured fluorescence.

  4. Stereoscopic Planar Laser-Induced Fluorescence Imaging at 500 kHz

    Science.gov (United States)

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

    2011-01-01

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

  5. [Laser Induced Fluorescence Spectroscopic Analysis of Aromatics from One Ring to Four Rings].

    Science.gov (United States)

    Zhang, Peng; Liu, Hai-feng; Yue, Zong-yu; Chen, Bei-ling; Yao, Ming-fa

    2015-06-01

    In order to distinguish small aromatics preferably, a Nd : YAG Laser was used to supply an excitation laser, which was adjusted to 0.085 J x cm(-2) at 266 nm. Benzene, toluene, naphthalene, phenanthrene, anthracene, pyrene and chrysene were used as the representative of different rings aromatics. The fluorescence emission spectra were researched for each aromatic hydrocarbon and mixtures by Laser induced fluorescence (LIF). Results showed that the rings number determined the fluorescence emission spectra, and the structure with same rings number did not affect the emission fluorescence spectrum ranges. This was due to the fact that the absorption efficiency difference at 266 nm resulted in that the fluorescence intensities of each aromatic hydrocarbon with same rings number were different and the fluorescence intensities difference were more apparently with aromatic ring number increasing. When the absorption efficiency was similar at 266 nm and the concentrations of each aromatic hydrocarbon were same, the fluorescence intensities were increased with aromatic ring number increasing. With aromatic ring number increasing, the fluorescence spectrum and emission peak wavelength were all red-shifted from ultraviolet to visible and the fluorescence spectrum range was also wider as the absorption efficiency was similar. The fluorescence emission spectra from one to four rings could be discriminated in the following wavelengths, 275 to 320 nm, 320 to 375 nm, 375 to 425 nm, 425 to 556 nm, respectively. It can be used for distinguish the type of the polycyclic aromatic hydrocarbons (PAHs) as it exists in single type. As PAHs are usually exist in a variety of different rings number at the same time, the results for each aromatic hydrocarbon may not apply to the aromatic hydrocarbon mixtures. For the aromatic hydrocarbon mixtures, results showed that the one- or two-ring PAHs in mixtures could not be detected by fluorescence as three- or four-ring PAHs existed in mixture

  6. Shrink-induced silica multiscale structures for enhanced fluorescence from DNA microarrays.

    Science.gov (United States)

    Sharma, Himanshu; Wood, Jennifer B; Lin, Sophia; Corn, Robert M; Khine, Michelle

    2014-09-23

    We describe a manufacturable and scalable method for fabrication of multiscale wrinkled silica (SiO2) structures on shrink-wrap film to enhance fluorescence signals in DNA fluorescence microarrays. We are able to enhance the fluorescence signal of hybridized DNA by more than 120 fold relative to a planar glass slide. Notably, our substrate has improved detection sensitivity (280 pM) relative to planar glass slide (11 nM). Furthermore, this is accompanied by a 30-45 times improvement in the signal-to-noise ratio (SNR). Unlike metal enhanced fluorescence (MEF) based enhancements, this is a far-field and uniform effect based on surface concentration and photophysical effects from the nano- to microscale SiO2 structures. Notably, the photophysical effects contribute an almost 2.5 fold enhancement over the concentration effects alone. Therefore, this simple and robust method offers an efficient technique to enhance the detection capabilities of fluorescence based DNA microarrays.

  7. Determination of phycobiliproteins by capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Viskari, P J; Kinkade, C S; Colyer, C L

    2001-07-01

    Phycobiliproteins are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae. They are composed of a protein backbone to which linear tetrapyrrole chromophores are covalently bound. Furthermore, they are water-soluble highly fluorescent, and relatively stable at room temperature and neutral pH. For this reason, capillary electrophoresis-laser induced fluorescence (CE-LIF) seems the idea method for determination of these important proteins. The effects of buffer additives such as sodium dodecyl sulfate (SDS)and putrescine on the separation of the three major phycobiliprotein types, namely allophycocyanin, phycocyanin, and phycoerythrin, with excitation and emission maxima at 652/660, 615/647, and 565(494)/575 nm, respectively, are considered. Detection limits for these proteins by CE-LIF are some 60-500 times better than by absorbance detection. The development of a fast and sensitive CE-LIF assay such as this is of potential significance to our understand ing of chemical and biological oceanographic processes.

  8. Application of Laser Induced Fluorescence in experimental analysis of convection phenomena

    Science.gov (United States)

    Jaszczur, M.; Pyrda, L.

    2016-09-01

    One of the most promising technique for temperature measurement is the Laser Induced Fluorescence (LIF) which utilize basic concept of optical, non-intrusive measurements and give possibility to visualize the temperature distribution in the whole two-dimensional plane at once. A major problem of a LIF is it still unsatisfactory accuracy for temperature gradient and heat transfer measurement. The LIF fluorescent re-emission is a function of temperature but, in all measurements a lot of imperfection follow the image recording: background noises, light intensity variation, non uniformity and shadowgraph effect near the non-isothermal walls. In the present paper the influence of all key effect on the temperature measurement will be verified in order to obtain the method uncertainty. To evaluate that experimental measurement of convection phenomenon using LIF and thermocouples focussing on heat transfer measurement will be presented. Results show that all processing steps (pre-processing, processing and post-processing) are crucial for reducing the error related to the temperature measurement.

  9. Portable detection system of vegetable oils based on laser induced fluorescence

    Science.gov (United States)

    Zhu, Li; Zhang, Yinchao; Chen, Siying; Chen, He; Guo, Pan; Mu, Taotao

    2015-11-01

    Food safety, especially edible oils, has attracted more and more attention recently. Many methods and instruments have emerged to detect the edible oils, which include oils classification and adulteration. It is well known than the adulteration is based on classification. Then, in this paper, a portable detection system, based on laser induced fluorescence, is proposed and designed to classify the various edible oils, including (olive, rapeseed, walnut, peanut, linseed, sunflower, corn oils). 532 nm laser modules are used in this equipment. Then, all the components are assembled into a module (100*100*25mm). A total of 700 sets of fluorescence data (100 sets of each type oil) are collected. In order to classify different edible oils, principle components analysis and support vector machine have been employed in the data analysis. The training set consisted of 560 sets of data (80 sets of each oil) and the test set consisted of 140 sets of data (20 sets of each oil). The recognition rate is up to 99%, which demonstrates the reliability of this potable system. With nonintrusive and no sample preparation characteristic, the potable system can be effectively applied for food detection.

  10. A simple dental caries detection system using full spectrum of laser-induced fluorescence

    Science.gov (United States)

    Rocha-Cabral, Renata Maciel; Mendes, Fausto Medeiros; Maldonado, Edison Puig; Zezell, Denise Maria

    2015-06-01

    Objectives: to develop an apparatus for the detection of early caries lesions in enamel using the full extent of the tooth fluorescence spectrum, through the integration of a laser diode, fiber optics, filters and one portable spectrometer connected to a computer, all commercially available; to evaluate the developed device in clinical and laboratory tests, and compare its performance with commercial equipment. Methods: clinical examinations were performed in patients with indication for exodontics of premolars. After examinations, the patients underwent surgery and the teeth were stored individually. The optical measurements were repeated approximately two months after extraction, on the same sites previously examined, then histological analysis was carried out. Results: the spectral detector has presented high specificity and moderate sensitivity when applied to differentiate between healthy and damaged tissues, with no significant differences from the performance of the commercial equipment. The developed device is able to detect initial damages in enamel, with depth of approximately 300 μm. Conclusions: we successfully demonstrated the development of a simple and portable system based in laser-induced fluorescence for caries detection, assembled from common commercial parts. As the spectral detector acquires a complete recording of the spectrum from each tissue, it is possible to use it for monitoring developments of caries lesions.

  11. Relationship between DAPI-fluorescence fading and nuclear DNA content: An alternative method to DNA quantification?

    Science.gov (United States)

    Gallardo-Escárate, Cristian; Alvarez-Borrego, Josué; Von Brand, Elisabeth; Dupré, Enrique; Del Río-Portilla, Miguel Angel

    2007-01-01

    In observations by confocal or conventional fluorescence microscopy, important factors should be considered in order to obtain accurate images. One of them, such as the fluorescence bleaching from highest intensity to lowest signal of fluorescence is a common problem with several DNA fluorochromes and especially for DAPI stain. The fluorescence of DAPI fades rapidly when it is exposed to UV light, under optimal conditions of observation. Although the fading process can be retarded using a mounting medium with antifading reagents, the photochemical process underlying the fluorescence decay has not yet been fully explained. In addition, no relationship between fluorescence fading and nuclear DNA content has been tested. In order to test this relationship, we measured by means of image analysis the DAPI-fluorescence intensity in several cellular types (spermatozoa, erythrocytes and haemocytes) during their fluorescence bleaching. An algorithm specifically built in MATLAB software was used for this approach. The correlation coefficient between nuclear DNA content and DAPI-fluorescence fading was found equal to 99%. This study demonstrates the feasibility to measure nuclear DNA content by fluorescence fading quantification, as an alternative method concurrently with image analysis procedures.

  12. [A fluorescence quenching method for the determination of trace chlorite in water with rhodamine B].

    Science.gov (United States)

    Kang, Cai-yan; Jiang, Zhi-liang; Xi, Dan-li

    2007-02-01

    In acidic sodium acetate-HCl buffer solution containing KI, Rhodamine B (RhB) has a fluorescence peak at 580 nm. When ClO2(-) exists fluorescence quenching occur. The fluorescence quenching intensity is linear with the concentration of ClO2(-) in the range of 0.0218-0.51 microg x mL(-1). Based on this, a new, simple, sentisive fluorescence method has been proposed for the determination of ClO2(-) in water, with satisfactory results.

  13. Directed evolution methods for improving polypeptide folding and solubility and superfolder fluorescent proteins generated thereby

    Science.gov (United States)

    Waldo, Geoffrey S.

    2007-09-18

    The current invention provides methods of improving folding of polypeptides using a poorly folding domain as a component of a fusion protein comprising the poorly folding domain and a polypeptide of interest to be improved. The invention also provides novel green fluorescent proteins (GFPs) and red fluorescent proteins that have enhanced folding properties.

  14. New method for covalent fluorescent biomolecule labeling with hemicyanine dye.

    Science.gov (United States)

    Kostenko, Olexander M; Kovalska, Vladyslava B; Volkova, Kateryna D; Shaytanov, Pavel; Kocheshev, Igor O; Slominskiy, Yuriy L; Pisareva, Irina V; Yarmoluk, Sergiy M

    2006-07-01

    Fluorescent chromophore, alkylamino-(tetra-hydronaphthalenylidene)- benzothiazolium derivatives (HBTN dyes), are proposed as covalent labels for proteins via aliphatic amino groups. Spectral-luminescent properties of 3-methyl-2-{(E)-[7-(methylamino)-4,4a,5,6-tetra-hydronaphthalen-2(3H)-ylidene]methyl}-1,3-benzothiazol-3-ium chloride (HBTN, R=Me) and its predecessor, 2-[(E)-(7-methoxy-4,4a,5,6-tetrahydronaphthalen-2(3H)-ylidene)methyl]-3-methyl-1,3-benzothiazol-3-ium chloride (ABTN), are studied for free dyes and in the presence of DNA and BSA. Considerable spectral-luminescent changes accompany the transformation of ABTN into HBTN that allows monitoring conjugation reaction. In presence of DNA and BSA the HBTN increases its emission in 15 and 4 times respectively and becomes strongly fluorescent. The conditions for labeling are developed and a model conjugate of HBTN dye with BSA is synthesized. It was shown that using of HBTN dye as a fluorescent label allows detection by eye of about 3 mug/band of BSA on polyacrylamide gel upon UV-irradiation.

  15. Laser induced fluorescence and Raman spectroscopy in capillary electrophoresis as an possible instrument for extraterrestrial life signs detection.

    Science.gov (United States)

    Mikhail, Gorlenko; Cheptcov, Vladimir; Anton, Maydykovskiy; Eugeniy, Vasilev

    The one of a significant aims in extraterrestrial exploration is a seeking for a life traces in a open space and planetary objects. Complex composition and unknown origin of suspected signs of life required у new analytical approaches and technical solutions. The promising assai here can be Laser induced fluorescence and Raman spectroscopy methods. The combined instrument developed by our team reveal the advantage of capillary electrophoresis assays in a junction with laser induced fluorescence detection technology. We optimized excitation configuration of fluorescence in capillary electrophoresis to reduce pumping laser power up to 1 mW and decrease background scattering. The improvement of the device sensitivity at poor sample concentration we achieved by incorporating fluorescence flow-through cuvette into spectrometer. That allows to simplify setup, to minimize weight and increase reproducibility of measurements. The device has been tasted in complex organic chemical mixes and microbial strains differentiation tasks. 3d multinational spectra allow us to increase the spectra information loads in comparison with ordinary capillary electrophoresis approaches. Possible updating the device with Raman approach can even furthermore multiple the differentiation power of the instrument. The analytical module developed using this approach can be potentially effectively used in extraterrestrial researches as a payload of the future spacecraft.

  16. Detection of fecal residue on poultry carcasses by laser-induced fluorescence imaging.

    Science.gov (United States)

    Cho, B; Kim, M S; Chao, K; Lawrence, K; Park, B; Kim, K

    2009-04-01

    Feasibility of fluorescence imaging technique for the detection of diluted fecal matters from various parts of the digestive tract, including colon, ceca, small intestine, and duodenum, on poultry carcasses was investigated. One of the challenges for using fluorescence imaging for inspection of agricultural material is the low fluorescence yield in that fluorescence can be masked by ambient light. A laser-induced fluorescence imaging system (LIFIS) developed by our group allowed acquisition of fluorescence from feces-contaminated poultry carcasses in ambient light. Fluorescence emission images at 630 nm were captured with 415-nm laser excitation. Image processing algorithms including threshold and image erosion were used to identify fecal spots diluted up to 1: 10 by weight with double distilled water. Feces spots on the carcasses, without dilution and up to 1: 5 dilutions, could be detected with 100% accuracy regardless of feces type. Detection accuracy for fecal matters diluted up to 1: 10 was 96.6%. The results demonstrated good potential of the LIFIS for detection of diluted poultry fecal matter, which can harbor pathogens, on poultry carcasses.

  17. Interaction between bovine serum albumin and Indo-1 using fluorescence spectroscopic method

    Institute of Scientific and Technical Information of China (English)

    Haixin BAI; Cheng YANG; Xiurong YANG

    2008-01-01

    This work attempts to calculate the binding-site number using fluorescence spectroscopic method with bovine serum albumin (BSA) and Indo-1 as proteinand ligand models, respectively. The method for calculat-ing the binding-site number in BSA for Indo-1 was developed based on the relationships between changes in Indo-1 fluorescence intensity and the analytical concen-tration of BSA. The interaction between BSA with Indo-1 was investigated comprehensively using fluorescence techniques as well as fluorescence resonance energy transfer, and the thermodynamic parameters were calculated according to the effect of enthalpy on temperature. Three binding sites in BSA for Indo-1 were revealed, and the distances from Trp212 in BSA to the three binding sites were 2.93, 2.57 and 2.40 nm, respectively. It was also proven that Indo-1 embedded into the three hydrophobic cavities of BSA by hydro-phobic association. This paper provides a reference on calculating the binding-site number in proteins for ligands and studying their interactions by fluorescence spectroscopic methods. In fluorescent quenching experi-ments, fluorescence changes were automatically recorded in real time by combining the Microlab 500 Series Dispenser and PTI fluorescence apparatus.

  18. Detection and mapping of oil-contaminated soils by remote sensing of laser-induced fluorescence

    Science.gov (United States)

    Luedeker, Wilhelm; Guenther, Kurt P.; Dahn, Hans-Guenter

    1995-10-01

    The contamination of soil by aromatic mineral hydrocarbons (MHC) (e.g., gasoline, oil, etc.) has become a severe environmental problem because not only men, animals, and plants are threatened but also the water and air. With the unification of Germany a great number of suspected contaminated sites in the new countries were registered. An estimation of the German Federal Ministry of Environment (BMU) counts 180,000 areas contaminated with different pollutants, 55,000 are situated in the former GDR. On military settlements for example more than fifty percent of the chemicals are MHCs. Hence one can get an idea of the importance of soil pollution by hydrocarbons. Other zones contaminated due to carelessness or accidents are civil petrolstations, airports, refineries, pipelines, and traffic disasters. At the present time for most of these areas the contamination is assumed due to recent use. Due to the large extension of the problem an estimation and evaluation of the potential hazard for the environment is difficult and expensive to perform. In the case of an actual endangering the total area must be mapped in detail resulting in increasing costs for the owner. Nevertheless it is necessary to find reliable timesaving areal mapping and monitoring methods. One opportunity presented in this paper is the application of remote sensing by laser induced fluorescence from an airborne platform. It promises to fulfill these requirements in a sufficiently fast manner with very high spatial resolution. The access to the pollutant detection is the specific laser induced fluorescence emitted by the MHC (finger print). The present work shows the requirements for a helicopterborne lidar system for MHC mapping and how the detected signals are to be evaluated and interpreted.

  19. The Three-region Method of Color Matching in the Presence of Fluorescent Whitening Agents

    Institute of Scientific and Technical Information of China (English)

    2005-01-01

    An expression of reflectance of fluorescent dyes in emissionregion was used in this article. Through the analysis of thesppectral radiance factor of fluorescent whitening agents (FWA), the color matching of FWA was studied and the three-region method of color matching method was put forward. The method was proved to be reliable for the dyes used and was easy to apply for the color matching of FWAs.

  20. Strategies of fluorescence staining for trace total ribonucleic acid analysis by capillary electrophoresis with argon ion laser-induced fluorescence.

    Science.gov (United States)

    Chung, Yi-An; Chen, Yi-Hsin; Chang, Po-Ling

    2015-08-01

    In this work, five fluorescent dyes (SYTO-9, SYBR Green I, SYBR Green II, SYBR Safe, and SYBR Gold) were used as both on-column and precolumn stains for total RNA analysis by CE-LIF with Ar ion laser excitation. In the on-column RNA stain, the SYTO-9 provided the highest fluorescence intensity and the lowest detectable concentration, as low as 10 pg/μL, while the SYBR Green II and SYBR Gold were adsorbed on the poly(ethylene oxide) thus affected the separation efficiency. As a precolumn stain, SYBR Gold was the most sensitive among the five dyes due to the strong affinity between the dye and RNA molecules. As a result, a single-cell quantity of RNA (10-30 pg per cell) could be detected by CE-LIF with precolumn staining by SYBR Gold. Because of the great savings of fluorescent dye using precolumn stain (one button dye may use for one million stain), this method is the best strategy for RNA staining in terms of cost-effectiveness and sensitivity.

  1. Investigation of fluorescence methods for rapid detection of municipal wastewater impact on drinking water sources

    Science.gov (United States)

    Peleato, Nicolas M.; Legge, Raymond L.; Andrews, Robert C.

    2017-01-01

    Fluorescence spectroscopy as a means to detect low levels of treated wastewater impact on two source waters was investigated using effluents from five wastewater facilities. To identify how best to interpret the fluorescence excitation-emission matrices (EEMs) for detecting the presence of wastewater, several feature selection and classification methods were compared. An expert supervised regional integration approach was used based on previously identified features which distinguish biologically processed organic matter including protein-like fluorescence and the ratio of protein to humic-like fluorescence. Use of nicotinamide adenine dinucleotide-like (NADH) fluorescence was found to result in higher linear correlations for low levels of wastewater presence. Parallel factors analysis (PARAFAC) was also applied to contrast an unsupervised multiway approach to identify underlying fluorescing components. A humic-like component attributed to reduced semiquinone-like structures was found to best correlate with wastewater presence. These fluorescent features were used to classify, by volume, low (0.1-0.5%), medium (1-2%), and high (5-15%) levels by applying support vector machines (SVMs) and logistic regression. The ability of SVMs to utilize high-dimensional input data without prior feature selection was demonstrated through their performance when considering full unprocessed EEMs (66.7% accuracy). The observed high classification accuracies are encouraging when considering implementation of fluorescence spectroscopy as a water quality monitoring tool. Furthermore, the use of SVMs for classification of fluorescence data presents itself as a promising novel approach by directly utilizing the high-dimensional EEMs.

  2. Development of a Fluorescence Quantitative PCR Method for Detection of Marteilia refringens in Shellfish

    Institute of Scientific and Technical Information of China (English)

    Liji XIE; Zhixun XIE; Yaoshan PANG; Jiabo LIU; Xianwen DENG; Zhiqin XIE

    2012-01-01

    Abstract [Objective] This paper was to develop a fluorescence quantitative PCR method for detection of M. refringens in shellfish. [Method] A pair of primers and a TaqMan probe were designed and synthesized according to the conserved gene se- quences of M. refringens in GenBank, so as to develop a fluorescence quantitative PCR method for detection of M. refringens. The developed fluorescence quantitative PCR method was compared with conventional PCR detection. [Result] The fluores- cence quantitative PCR could detect 40 template copies of plasmid DNA, and its sensitivity was 100 times higher than the conventional PCR. The detection results of Perkinsus sp, Haplosporidium sp, Aeromonas hydrophila, Pseudomonas fluorescens, Vibrio parahaemolyticu, Vibrio alginolyticu, Vibrio rluvialis and Vibrio mimicus were negtive. [Conclusion] The fluorescence quantitative PCR method for M. refringens es- tablished in this paper is specific, sensitive, rapid and quantitative with good re- peatability, which can be used for clinical detection of M. refringens infection.

  3. [Study of the Detecting System of CH4 and SO2 Based on Spectral Absorption Method and UV Fluorescence Method].

    Science.gov (United States)

    Wang, Shu-tao; Wang, Zhi-fang; Liu, Ming-hua; Wei, Meng; Chen, Dong-ying; Wang, Xing-long

    2016-01-01

    According to the spectral absorption characteristics of polluting gases and fluorescence characteristics, a time-division multiplexing detection system is designed. Through this system we can detect Methane (CH4) and sulfur dioxide (SO2) by using spectral absorption method and the SO2 can be detected by using UV fluorescence method. The system consists of four parts: a combination of a light source which could be switched, the common optical path, the air chamber and the signal processing section. The spectral absorption characteristics and fluorescence characteristics are measured first. Then the experiment of detecting CH4 and SO2 through spectral absorption method and the experiment of detecting SO2 through UV fluorescence method are conducted, respectively. Through measuring characteristics of spectral absorption and fluorescence, we get excitation wavelengths of SO2 and CH4 measured by spectral absorption method at the absorption peak are 280 nm and 1.64 μm, respectively, and the optimal excitation wavelength of SO2 measured by UV fluorescence method is 220 nm. we acquire the linear relation between the concentration of CH4 and relative intensity and the linear relation between the concentration of SO2 and output voltage after conducting the experiment of spectral absorption method, and the linearity are 98.7%, 99.2% respectively. Through the experiment of UV fluorescence method we acquire that the relation between the concentration of SO2 and the voltage is linear, and the linearity is 99.5%. Research shows that the system is able to be applied to detect the polluted gas by absorption spectrum method and UV fluorescence method. Combing these two measurement methods decreases the costing and the volume, and this system can also be used to measure the other gases. Such system has a certain value of application.

  4. Detection of bacterial infection of agave plants by laser-induced fluorescence

    Science.gov (United States)

    Cervantes-Martinez, Jesus; Flores-Hernandez, Ricardo; Rodriguez-Garay, Benjamin; Santacruz-Ruvalcaba, Fernando

    2002-05-01

    Greenhouse-grown plants of Agave tequilana Weber var. azul were inoculated with Erwinia carotovora, the causal agent of stem soft rot. We investigated the laser-induced fluorescence (LIF) of agave plants to determine whether LIF can be used as a noninvasive sensing tool for pathological studies. The LIF technique was also investigated as a means of detecting the effect of the polyamine biosynthesis inhibitor beta-hydroxyethylhydrazine as a bactericide against the pathogenic bacterium Erwinia carotovora. A He-Ne laser at 632.8 nm was used as the excitation source, and in vivo fluorescence emission spectra were recorded in the 660-790-range. Fluorescence maxima were at 690 and 740 nm. The infected plants that were untreated with the bactericide showed a definite increase in fluorescence intensity at both maxima within the first three days after infection. Beginning on the fifth day, a steady decrease in fluorescence intensity was observed, with a greater effect at 740 than at 690 nm. After 30 days there was no fluorescence. The infected plants that had been treated with the bactericide showed no significant change in fluorescence compared with that of the uninfected plants. The ratio of fluorescence intensities was determined to be F 690 nm/F 740 nm for all treatments. These studies indicate that LIF measurements of agave plants may be used for the early detection of certain types of disease and for determining the effect of a bactericide on bacteria. The results also showed that fluorescence intensity ratios can be used as a reliable indicator of the progress of disease.

  5. Quantification of Canine Dental Plaque Using Quantitative Light-Induced Fluorescence.

    Science.gov (United States)

    Wallis, Corrin; Gill, Yadvinder; Colyer, Alison; Davis, Ian; Allsopp, Judi; Komarov, Gleb; Higham, Susan; Harris, Stephen

    2016-03-01

    The aim of this work was to evaluate Quantitative Light-induced Fluorescence (QLF) as an alternative to the established Logan and Boyce method for determining plaque coverage of dogs' teeth. In a series of studies in conscious and anesthetized dogs, QLF showed good intra-photographer repeatability (coefficient of variation [CV] of 7.5% for undisclosed teeth) and inter-photographer reproducibility (CV of 3.2% for undisclosed teeth and 8.5% for disclosed teeth). The QLF software accurately identifies areas of plaque as demonstrated by comparison to the variability of 5 human scorers, manually marking plaque on QLF-acquired images (P = 0.1). There was good agreement with the modified Logan and Boyce method in the percentage reduction in plaque accumulation measured when dogs were fed an oral care chew versus no chew. To see a 15% difference in plaque accumulation, which is considered sufficient by the Veterinary Oral Health Council to differentiate between 2 treatments, a retrospective power analysis (90%) of the data established that only 7 dogs would be required, compared to 19 dogs for the modified Logan and Boyce method. QLF is a reliable method for measuring dental plaque in dogs with the added advantage that it is not subjective and requires fewer animals.

  6. Planar laser-induced fluorescence fuel imaging during gas-turbine relight

    DEFF Research Database (Denmark)

    Read, Robert; Rogerson, J.W.; Hochgreb, S.

    2013-01-01

    This experimental study investigates the influence of fuel distribution on ignition outcome during high-altitude relight of a gas turbine. Planar laser-induced fluorescence is used to image fuel inside a lean direct-injection combustor under realistic conditions. A novel apparatus is developed...... to permit planar laser-induced fluorescence imaging, in which large quantities of poorly atomized fuel impinges on the internal surfaces of the combustor. Results reveal high variability in atomization quality. In the absence of flame, small droplets are confined to areas of recirculating flow, whereas...... of ignition when the airflow rate is high. In the presence of flame, medium-sized burning droplets are observed close to the injector centerline. Flame interference resulting from fluorescence of polycyclic aromatic hydrocarbons is apparent, but small, suggesting that kerosene planar laser...

  7. Embryonic and induced pluripotent stem cell staining and sorting with the live-cell fluorescence imaging probe CDy1.

    Science.gov (United States)

    Kang, Nam-Young; Yun, Seong-Wook; Ha, Hyung-Ho; Park, Sung-Jin; Chang, Young-Tae

    2011-06-30

    Detecting and isolating specific types of cells is crucial to understanding a variety of biological processes, including development, aging, regeneration and pathogenesis; this understanding, in turn, allows the use of cells for therapeutic purposes, for which stem cells have emerged recently as invaluable materials. The current methods of isolation and characterization of stem cells depend on cell morphology in culture or on immunostaining of specific markers. These methods are, however, time consuming and involve the use of antibodies that may often make the cells unsuitable for further study. We recently developed a fluorescent small molecule named CDy1 (compound of designation yellow 1) that selectively stains live embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs). This protocol describes detailed procedures for staining ESC and iPSC in live conditions and for fluorescence-activated cell sorting (FACS) of ESC using CDy1. Cell staining, image acquisition and FACS can be done within 6 h.

  8. Estimating chlorophyll content and photochemical yield of photosystem II (ΦPSII) using solar-induced chlorophyll fluorescence measurements at different growing stages of attached leaves.

    Science.gov (United States)

    Tubuxin, Bayaer; Rahimzadeh-Bajgiran, Parinaz; Ginnan, Yusaku; Hosoi, Fumiki; Omasa, Kenji

    2015-09-01

    This paper illustrates the possibility of measuring chlorophyll (Chl) content and Chl fluorescence parameters by the solar-induced Chl fluorescence (SIF) method using the Fraunhofer line depth (FLD) principle, and compares the results with the standard measurement methods. A high-spectral resolution HR2000+ and an ordinary USB4000 spectrometer were used to measure leaf reflectance under solar and artificial light, respectively, to estimate Chl fluorescence. Using leaves of Capsicum annuum cv. 'Sven' (paprika), the relationships between the Chl content and the steady-state Chl fluorescence near oxygen absorption bands of O2B (686nm) and O2A (760nm), measured under artificial and solar light at different growing stages of leaves, were evaluated. The Chl fluorescence yields of ΦF 686nm/ΦF 760nm ratios obtained from both methods correlated well with the Chl content (steady-state solar light: R(2) = 0.73; artificial light: R(2) = 0.94). The SIF method was less accurate for Chl content estimation when Chl content was high. The steady-state solar-induced Chl fluorescence yield ratio correlated very well with the artificial-light-induced one (R(2) = 0.84). A new methodology is then presented to estimate photochemical yield of photosystem II (ΦPSII) from the SIF measurements, which was verified against the standard Chl fluorescence measurement method (pulse-amplitude modulated method). The high coefficient of determination (R(2) = 0.74) between the ΦPSII of the two methods shows that photosynthesis process parameters can be successfully estimated using the presented methodology.

  9. Formation of nitric oxide in an industrial burner measured by 2-D laser induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Arnold, A.; Bombach, R.; Kaeppeli, B. [Paul Scherrer Inst. (PSI), Villigen (Switzerland)

    1997-06-01

    We have performed two-dimensional Laser Induced Fluorescence (2-D LIF) measurements of nitric oxide and hydroxyl radical distributions in an industrial burner at atmospheric pressure. The relative 2-D LIF data of NO were set to an absolute scale by calibration with probe sampling combined with gas analysis. (author) 3 figs., 7 refs.

  10. Capillary electrophoresis with laser-induced fluorescence detection for fast and reliable apolipoprotein E genotyping

    NARCIS (Netherlands)

    Somsen, GW; Welten, HTME; Mulder, FP; Swart, CW; Kema, IP; de Jong, GJ

    2002-01-01

    The use of capillary electrophoresis (CE) with laser-induced fluorescence (LIF) detection for the rapid determination of apolipoprotein E (apoE) genotypes was studied. High resolution and sensitive detection of the concerned DNA restriction fragments was achieved using CE buffers with hydroxypropylm

  11. Capillary Electrophoretic Immunoassay with Laser-induced Fluorescence Detection for Interferon-gamma

    Institute of Scientific and Technical Information of China (English)

    Hua ZHANG; Hai Ming WEI; Wen Rui JIN

    2004-01-01

    Capillary electrophoretic immunoassay with laser-induced fluorescence detection for recombinant human interferon-gamma (IFN-γ) was established. The limits of detection for three forms of IFN-γare 6.9 ng/L, 5.7 ng/L and 5.0 ng/L, respectively.

  12. Two dye combinations suitable for two-color/two-dye laser-induced fluorescence thermography for ethanol

    Science.gov (United States)

    Fenner, Andreas; Stephan, Peter

    2017-06-01

    This paper presents two dye combinations suitable for two-color/two-dye laser-induced fluorescence thermography for ethanol. Besides the temperature dependency of the fluorescence, the influences of laser fluence, dye concentration, pressure, dissolved air, and photobleaching are also discussed. The experimental data are compared with models and data available in literature. Based on this, parameter ranges for two-color/two-dye laser-induced fluorescence thermography applications can be determined.

  13. Flow method and apparatus for screening chemicals using micro x-ray fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Warner, Benjamin P. (Los Alamos, NM); Havrilla, George J. (Los Alamos, NM); Miller, Thomasin C. (Bartlesville, OK); Lewis, Cris (Los Alamos, NM); Mahan, Cynthia A. (Los Alamos, NM); Wells, Cyndi A. (Los Alamos, NM)

    2011-04-26

    Method and apparatus for screening chemicals using micro x-ray fluorescence. A method for screening a mixture of potential pharmaceutical chemicals for binding to at least one target binder involves flow separating a solution of chemicals and target binders into separated components, exposing them to an x-ray excitation beam, detecting x-ray fluorescence signals from the components, and determining from the signals whether or not a binding event between a chemical and target binder has occurred.

  14. Flow method and apparatus for screening chemicals using micro x-ray fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Warner, Benjamin P. (Los Alamos, NM); Havrilla, George J. (Los Alamos, NM); Miller, Thomasin C. (Bartlesville, OK); Lewis, Cris (Los Alamos, NM); Mahan, Cynthia A. (Los Alamos, NM); Wells, Cyndi A. (Los Alamos, NM)

    2009-04-14

    Method and apparatus for screening chemicals using micro x-ray fluorescence. A method for screening a mixture of potential pharmaceutical chemicals for binding to at least one target binder involves flow-separating a solution of chemicals and target binders into separated components, exposing them to an x-ray excitation beam, detecting x-ray fluorescence signals from the components, and determining from the signals whether or not a binding event between a chemical and target binder has occurred.

  15. A novel method for the absolute fluorescence yield measurement by AIRFLY

    CERN Document Server

    Ave, M

    2008-01-01

    One of the goals of the AIRFLY (AIR FLuorescence Yield) experiment is to measure the absolute fluorescence yield induced by electrons in air to better than 10% precision. We introduce a new technique for measurement of the absolute fluorescence yield of the 337 nm line that has the advantage of reducing the systematic uncertainty due to the detector calibration. The principle is to compare the measured fluorescence yield to a well known process - the Cerenkov emission. Preliminary measurements taken in the BFT (Beam Test Facility) in Frascati, Italy with 350 MeV electrons are presented. Beam tests in the Argonne Wakefield Accelerator at the Argonne National Laboratory, USA with 14 MeV electrons have also shown that this technique can be applied at lower energies.

  16. Laser-induced fluorescence: quantitative analysis of atherosclerotic plaque chemical content in human aorta

    Science.gov (United States)

    Dai, Erbin; Wishart, David; Khoury, Samir; Kay, Cyril M.; Jugdutt, Bodh I.; Tulip, John; Lucas, Alexandra

    1996-05-01

    We have been studying laser-induced fluorescence as a technique for identification of selected changes in the chemical composition of atherosclerotic plaque. Formulae for quantification of chemical changes have been developed based upon analysis of fluorescence emission spectra using multiple regression analysis and the principal of least squares. The intima of human aortic necropsy specimens was injected with chemical compounds present in atherosclerotic plaque. Spectra recorded after injection of selected chemical components found in plaque (collagen I, III, IV, elastin and cholesterol) at varying concentrations (0.01 - 1.0 mg) were compared with saline injection. A single fiber system was used for both fluorescence excitation (XeCl excimer laser, 308 nm, 1.5 - 2.0 mJ/ pulse, 5 Hz) and fluorescence emission detection. Average spectra for each chemical have been developed and the wavelengths of peak emission intensity identified. Curve fitting analysis as well as multiple regression analysis were used to develop formulae for assessment of chemical content. Distinctive identifying average curves were established for each chemical. Excellent correlations were identified for collagen I, III, and IV, elastin, and cholesterol (R2 equals 0.92 6- 0.997). Conclusions: (1) Fluorescence spectra of human aortas were significantly altered by collagen I, collagen III, elastin and cholesterol. (2) Fluorescence spectroscopic analysis may allow quantitative assessment of atherosclerotic plaque chemical content in situ.

  17. [The analysis of sinusoidal modulated method used for measuring fluorescence lifetime].

    Science.gov (United States)

    Feng, Ying; Huang, Shi-hua

    2007-12-01

    This paper has built a system with a sinusoidal modulated LED as the excitation source. Such exciter was used upon the sample Eu2 L'3 x nH2O (L' = C4H4O4). Both the excitation light and the 5Do-7F2 emission of Eu3+ ion were measured. Fluorescence lifetime, which approximate to 0.680 ms, can then be obtained from the measured excitation and fluorescence waveforms by non-linear least square curve fitting based on the principle of phase-shift measurement of fluorescence lifetime. Data processing methods considering respectively the high order harmonics in the modulation and multi-exponential decay of the fluorescence were discussed. A method of utilizing Fourier series expandedness to amendatory the result was put forward. Accordingly, the applicability for phase-shift method was expanded as well as a more exact result was acquired.

  18. Fluorescence lifetime imaging of induced pluripotent stem cells

    Science.gov (United States)

    Uchugonova, Aisada; Batista, Ana; König, Karsten

    2014-02-01

    The multiphoton FLIM tomograph MPTflex with its flexible scan head, articulated arm, and the tunable femtosecond laser source was employed to study cell monolayers and 3D cell clusters. FLIM was performed with 250 ps temporal resolution and submicron special resolution using time-correlated single photon counting. The autofluorescence based on NAD(P)H and flavins/flavoproteins has been measured in mouse embryonic fibroblasts, induced pluripotent stem cells (iPS cells) originated from mouse embryonic fibroblasts and non-proliferative mouse embryonic fibroblasts.

  19. Laser-induced fluorescence reader with a turbidimetric system for sandwich-type immunoassay using nanoparticles

    Energy Technology Data Exchange (ETDEWEB)

    Kim, Y.H.; Lim, H.B., E-mail: plasma@dankook.ac.kr

    2015-07-09

    Graphical abstract: Laser-induced fluorescence reader with ratiometric correction for sandwich-type immunoassay using nanoparticles. - Highlights: • Laser-induced fluorescence system with ratiometric correction was developed. • The system reduced experimental error caused by particle loss and aggregation. • The detection limit of about 39 pg mL{sup −1} for salinomycin was obtained. • Calibration linearity and sensitivity were also significantly improved. • The system has the potential for bioanalysis using various nanoparticles. - Abstract: A unique laser-induced fluorescence (LIF) reader equipped with a turbidimetric system was developed for a sandwich-type immunoassay using nanoparticles. The system was specifically designed to reduce experimental error caused by particle loss, aggregation and sinking, and to improve analytical performance through ratiometric measurement of the fluorescence with respect to the turbidimetric absorbance. For application to determine the concentration of salinomycin, magnetic nanoparticles (MNPs) and FITC-doped silica nanoparticles (colored balls) immobilized with antibody were synthesized for magnetic extraction and for tagging as a fluorescence probe, respectively. The detection limit of about 39 pg mL{sup −1} was obtained, which was an improvement of about 2-fold compared to that obtained without employment of the turbidimetric system. Calibration linearity and sensitivity were also improved, with increase from 0.8601 to 0.9905 in the R{sup 2}-coefficient and by 1.92-fold for the curve slope, respectively. The developed LIF reader has the potential to be used for fluorescence measurements using various nanomaterials, such as quantum dots.

  20. Amine Analysis Using AlexaFluor 488 Succinimidyl Ester and Capillary Electrophoresis with Laser-Induced Fluorescence

    Directory of Open Access Journals (Sweden)

    Christian G. Kendall

    2015-01-01

    Full Text Available Fluorescent probes enable detection of otherwise nonfluorescent species via highly sensitive laser-induced fluorescence. Organic amines are predominantly nonfluorescent and are of analytical interest in agricultural and food science, biomedical applications, and biowarfare detection. Alexa Fluor 488 N-hydroxysuccinimidyl ester (AF488 NHS-ester is an amine-specific fluorescent probe. Here, we demonstrate low limit of detection of long-chain (C9 to C18 primary amines and optimize AF488 derivatization of long-chain primary amines. The reaction was found to be equally efficient in all solvents studied (dimethylsulfoxide, ethanol, and N,N-dimethylformamide. While an organic base (N,N-diisopropylethylamine is required to achieve efficient reaction between AF488 NHS-ester and organic amines with longer hydrophobic chains, high concentrations (>5 mM result in increased levels of ethylamine and propylamine in the blank. Optimal incubation times were found to be >12 hrs at room temperature. We present an initial capillary electrophoresis separation for analysis using a simple micellar electrokinetic chromatography (MEKC buffer consisting of 12 mM sodium dodecylsulfate (SDS and 5 mM carbonate, pH 10. Limits of detection using the optimized labeling conditions and these separation conditions were 5–17 nM. The method presented here represents a novel addition to the arsenal of fluorescent probes available for highly sensitive analysis of small organic molecules.

  1. In vitro cytotoxicity of fluorescent silica nanoparticles hybridized with aggregation-induced emission luminogens for living cell imaging.

    Science.gov (United States)

    Xia, Yun; Li, Min; Peng, Tao; Zhang, Weijie; Xiong, Jun; Hu, Qinggang; Song, Zifang; Zheng, Qichang

    2013-01-07

    Fluorescent silica nanoparticles (FSNPs) can provide high-intensity and photostable fluorescent signals as a probe for biomedical analysis. In this study, FSNPs hybridized with aggregation-induced emission (AIE) luminogens (namely FSNP-SD) were successfully fabricated by a surfactant-free sol-gel method. The FSNP-SD were spherical, monodisperse and uniform in size, with an average diameter of approximately 100 nm, and emitted strong fluorescence at the peak of 490 nm. The FSNP-SD selectively stained the cytoplasmic regions and were distributed in the cytoplasm. Moreover, they can stay inside cells, enabling the tacking of cells over a long period of time. The intracellular vesicles and multinucleated cells were increase gradually with the rise of FSNP-SD concentration. Both cell viability and survival only lost less than 20% when the cells were exposed to the high concentration of 100 μg/mL FSNP-SD. Additionally, the cell apoptosis and intracellular ROS assay indicated that FSNP-SD had no significant toxic effects at the maximum working concentration of 80 μg/mL. This study demonstrated that the FSNP-SD are promising biocompatible fluorescent probes for living cell imaging.

  2. In Vitro Cytotoxicity of Fluorescent Silica Nanoparticles Hybridized with Aggregation-Induced Emission Luminogens for Living Cell Imaging

    Directory of Open Access Journals (Sweden)

    Yun Xia

    2013-01-01

    Full Text Available Fluorescent silica nanoparticles (FSNPs can provide high-intensity and photostable fluorescent signals as a probe for biomedical analysis. In this study, FSNPs hybridized with aggregation-induced emission (AIE luminogens (namely FSNP-SD were successfully fabricated by a surfactant-free sol-gel method. The FSNP-SD were spherical, monodisperse and uniform in size, with an average diameter of approximately 100 nm, and emitted strong fluorescence at the peak of 490 nm. The FSNP-SD selectively stained the cytoplasmic regions and were distributed in the cytoplasm. Moreover, they can stay inside cells, enabling the tacking of cells over a long period of time. The intracellular vesicles and multinucleated cells were increase gradually with the rise of FSNP-SD concentration. Both cell viability and survival only lost less than 20% when the cells were exposed to the high concentration of 100 μg/mL FSNP-SD. Additionally, the cell apoptosis and intracellular ROS assay indicated that FSNP-SD had no significant toxic effects at the maximum working concentration of 80 μg/mL. This study demonstrated that the FSNP-SD are promising biocompatible fluorescent probes for living cell imaging.

  3. Laser-induced absorption and fluorescence studies of photochromic Schiff bases

    DEFF Research Database (Denmark)

    Kownacki, K.; Mordzinski, A.; Wilbrandt, R.

    1994-01-01

    Three photochromic Schiff bases: N-salicylideneaniline (SA), N-salicylidene-1-naphthylamine (SN), and N,N-bis-(salicylidene)-p-phenylenediamine (Bsp), were studied in acetonitrile by means of steady-state and time-resolved absorption and fluorescence spectroscopy, as well as semiempirical quantum...... chemical calculations. In all these molecules, the transient absorption and two-step laser-induced fluorescence spectra of long-lived transients are remarkably similar. The photochromic species is tentatively assigned to the non-hydrogen bonded form of the proton transfer reaction product...

  4. Differential Laser-Induced Perturbation Spectroscopy for Analysis of Mixtures of the Fluorophores l-Phenylalanine, l-Tyrosine and l-Tryptophan Using a Fluorescence Probe.

    Science.gov (United States)

    Oztekin, Erman K; Hahn, David W

    2016-09-01

    Quantitative detection of common endogenous fluorophores is accomplished using differential laser-induced perturbation spectroscopy (DLIPS) with a 193-nm UV fluorescence probe and various UV perturbation wavelengths. In this study, DLIPS is explored as an alternative to traditional fluorescence spectroscopy alone, with a goal of exploring natural fluorophores pursuant to biological samples and tissue analysis. To this end, aromatic amino acids, namely, l-phenylalanine, l-tyrosine and l-tryptophan are mixed with differing mass ratios and then classified with various DLIPS schemes. Classification with a traditional fluorescence probe is used as a benchmark. The results show a 20% improvement in classification performance of the DLIPS method over the traditional fluorescence method using partial least squares (PLS) analysis. Additional multivariate analyses are explored, and the relevant photochemistry is elucidated in the context of perturbation wavelengths. We conclude that DLIPS is a promising biosensing approach with potential for in vivo analysis given the current findings with fluorophores relevant to biological tissues.

  5. A Spectral Analysis of Laser Induced Fluorescence of Iodine

    CERN Document Server

    Bayram, S B

    2015-01-01

    When optically excited, iodine absorbs in the 490- to 650-nm visible region of the spectrum and, after radiative relaxation, it displays an emission spectrum of discrete vibrational bands at moderate resolution. This makes laser-induced fuorescence spectrum of molecular iodine especially suitable to study the energy structure of homonuclear diatomic molecules at room temperature. In this spirit, we present a rather straightforward and inexpensive experimental setup and the associated spectral analysis which provides an excellent exercise of applied quantum mechanics fit for advanced laboratory courses. The students would be required to assign spectral lines, fill a Deslandres table, process the data to estimate the harmonic and anharmonic characteristics of the ground vibronic state involved in the radiative transitions, and thenceforth calculate a set of molecular constants and discuss a model of molecular vibrator.

  6. Parameter Optimization of Information Channels for Laser Fluorescence Method of Vegetation Monitoring

    Directory of Open Access Journals (Sweden)

    M. L. Belov

    2015-01-01

    Full Text Available Nowadays, there is a growing interest in application of remote monitoring and accounting systems in agriculture.One of the promising areas of remote vegetation monitoring is a fluorescence analysis, as it potentially allows sensing stress of plants according to characteristics of their fluorescent radiation.The shape of the fluorescence spectra of vegetation in the normal condition differs from that of the fluorescence spectra of vegetation in stressful conditions. This potentially allows you to sence the plants by recording information about the shape of the fluorescence spectra.Analysis of the fluorescence spectrum shape can be replaced by the analysis of fluorescence intensities in several spectral bands, which simpifies problem-solving.Currently, there are various devices developed for laser fluorescence sensing of plant stress. However, a lot of issues important to the practice remain unclear.Most of these issues concern the parameters of receiving channels to record information signals, which allow you to perceve the stress-sensed plants:- how many information channels of spectral bands better to use;- what the best width of these spectral bands of information is ;- what is the best width of the spectral bands of information;- what the best threshold value for the threshold algorithm is, and if there is the better algorithm to process measurement data.The work uses mathematical modeling based on the experimentally measured fluorescence spectra to determine the optimal (in terms of probability of sensing characteristics of the stress of plants, i.e. the probability of good sense and false alarm parameters of information channels for laser fluorescence method to sense the plant stress: the central wavelength of the information spectral bands, their spectral width, and parameters of the algorithm in the case of processing two spectral channels of information. It is shown that using the additional third information spectral band allows you to

  7. Neisseria gonorrhoeae Identification in Direct Smears by a Fluorescent Antibody-Counterstain Method

    Science.gov (United States)

    White, Lendell A.; Kellogg, Douglas S.

    1965-01-01

    Direct smears from female patients have been considered unreliable for the detection of Neisseria gonorrhoeae by fluorescent-antibody (FA) methods because of inadequate background contrast of the fluorescent-stained smears and a scarcity of organisms on the smear. Evans blue dye employed as a counterstain eliminated the nonspecific background staining and increased the reliability of the direct FA procedure. Direct smears demonstrating positive fluorescence were obtained from 86% of a group of culturally positive named female contacts. The FA-counterstain technique is as sensitive as the presently recommended cultural procedures. PMID:14325874

  8. LED-Induced Fluorescence System for Tea Classification and Quality Assessment

    CERN Document Server

    Dong, Yongjiang; Mei, Liang; Feng, Chao; Yan, Chunsheng; He, Sailing

    2013-01-01

    A fluorescence system is developed by using several light emitting diodes (LEDs) with different wavelengths as excitation light sources. The fluorescence detection head consists of multi LED light sources and a multimode fiber for fluorescence collection, where the LEDs and the corresponding filters can be easily chosen to get appropriate excitation wavelengths for different applications. By analyzing fluorescence spectra with the principal component analysis method, the system is utilized in the classification of four types of green tea beverages and two types of black tea beverages. Qualities of the Xihu Longjing tea leaves of different grades, as well as the corresponding liquid tea samples, are studied to further investigate the ability and application of the system in the evaluation of classification/quality of tea and other foods.

  9. Chiral separation of benzoporphyrin derivative mono- and diacids by laser induced fluorescence-capillary electrophoresis.

    Science.gov (United States)

    Peng, Xuejun; Sternberg, Ethan; Dolphin, David

    2002-01-01

    A method for the separation of benzoporphyrin derivative mono- and diacid (BPDMA, BPDDA) enantiomers by laser induced fluorescence-capillary electrophoresis (LIF-CE) has been developed. By using 300 mM borate buffer, pH 9.2, 25 mM sodium cholate and 10% acetronitrile as electrolyte, +10 kV electrokinetic sampling injection of 2 s and an applied +20 kV voltage across the ends of a 37 cm capillary (30 cm to the detector, 50 microm ID), all six BPD stereoisomers were baseline-separated within 20 min. Formation constants, free electrophoretic and complexation mobilities with borate and cholate were determined based on dynamic complexation capillary electrophoresis theory. The BPD enantiomers can be quantitatively determined in the range of 10(-2)-10(-5) mg mL(-1). The correlation coefficients (r2) of the least-squares linear regression analysis of the BPD enantiomers are in the range of 0.9914-0.9997. Their limits of detection are 2.18-3.5 x 10(-3) mg mL(-1). The relative standard deviations for the separation were 2.90-4.64% (n = 10). In comparison with high-performance liquid chromatography (HPLC), CE has better resolution and efficiency. This separation method was successfully applied to the BPD enantiomers obtained from a matrix of bovine serum and from liposomally formulated material as well as from studies with rat, dog and human microsomes.

  10. Hot-wire accuracy in supersonic turbulence from comparisons with laser-induced fluorescence

    Science.gov (United States)

    Logan, Pamela; Bershader, Daniel; Mckenzie, Robert L.

    1988-01-01

    A hot-wire anemometer and a new, nonintrusive, laser-induced fluorescence (LIF) technique are used to survey a turbulent boundary layer in a supersonic channel flow at Mach no. 2.06. The purpose is to test the accuracy of using the hot wire to measure the fluctuation amplitudes of static temperature and density in a compressible turbulent flow by comparing the results with independent and direct LIF measurements. Several methods of hot-wire calibration and analysis are applied. With each method, the hot-wire response can be related primarily to fluctuations of mass flux and total temperature, from which fluctuations of static temperature and density are calculated. However, these calculations are shown to be valid only if the fluctuations in static pressure are negligible. The acquisition and the analysis of the hot-wire data are often simplified further by neglecting the effects of fluctuations in total temperature. Comparisons of the fluctuation amplitudes of temperature and density obtained by hot-wire and LIF measurements demonstrate that such assumptions might not always be warranted, even in apparently simple flows.

  11. Advanced fluorescence microscopy methods for the real-time study of transcription and chromatin dynamics.

    Science.gov (United States)

    Annibale, Paolo; Gratton, Enrico

    2014-03-12

    In this contribution we provide an overview of the recent advances allowed by the use of fluorescence microscopy methods in the study of transcriptional processes and their interplay with the chromatin architecture in living cells. Although the use of fluorophores to label nucleic acids dates back at least to about half a century ago, (1) two recent breakthroughs have effectively opened the way to use fluorescence routinely for specific and quantitative probing of chromatin organization and transcriptional activity in living cells: namely, the possibility of labeling first the chromatin loci and then the mRNA synthesized from a gene using fluorescent proteins. In this contribution we focus on methods that can probe rapid dynamic processes by analyzing fast fluorescence fluctuations.

  12. Cyanine-based probe\\tag-peptide pair for fluorescence protein imaging and fluorescence protein imaging methods

    Science.gov (United States)

    Mayer-Cumblidge, M. Uljana; Cao, Haishi

    2010-08-17

    A molecular probe comprises two arsenic atoms and at least one cyanine based moiety. A method of producing a molecular probe includes providing a molecule having a first formula, treating the molecule with HgOAc, and subsequently transmetallizing with AsCl.sub.3. The As is liganded to ethanedithiol to produce a probe having a second formula. A method of labeling a peptide includes providing a peptide comprising a tag sequence and contacting the peptide with a biarsenical molecular probe. A complex is formed comprising the tag sequence and the molecular probe. A method of studying a peptide includes providing a mixture containing a peptide comprising a peptide tag sequence, adding a biarsenical probe to the mixture, and monitoring the fluorescence of the mixture.

  13. Quantitative two-dimensional measurement of oil-film thickness by laser-induced fluorescence in a piston-ring model experiment.

    Science.gov (United States)

    Wigger, Stefan; Füßer, Hans-Jürgen; Fuhrmann, Daniel; Schulz, Christof; Kaiser, Sebastian A

    2016-01-10

    This paper describes advances in using laser-induced fluorescence of dyes for imaging the thickness of oil films in a rotating ring tribometer with optical access, an experiment representing a sliding piston ring in an internal combustion engine. A method for quantitative imaging of the oil-film thickness is developed that overcomes the main challenge, the accurate calibration of the detected fluorescence signal for film thicknesses in the micrometer range. The influence of the background material and its surface roughness is examined, and a method for flat-field correction is introduced. Experiments in the tribometer show that the method yields quantitative, physically plausible results, visualizing features with submicrometer thickness.

  14. Transient isotachophoretic-electrophoretic separations of lanthanides with indirect laser-induced fluorescence detection.

    Science.gov (United States)

    Church, M N; Spear, J D; Russo, R E; Klunder, G L; Grant, P M; Andresen, B D

    1998-07-01

    Indirect laser-induced fluorescence was used for the detection of several lanthanide species separated by capillary electrophoresis. Quinine sulfate was the fluorescent component of the background electrolyte, and α-hydroxyisobutyric acid was added as a complexing agent to enable the separation of analyte ions that have similar mobilities. The UV lines (333-364 nm) of an argon ion laser were used as the excitation source with a diode array detector for monitoring the fluorescent emission at 442 nm. Electrokinetic injections and transient isotachophoresis were implemented to stack the analyte ions into more concentrated zones. On-line preconcentration factors were determined to be ∼700 and resulted in limits of detection for La(3+), Ce(3+), Pr(3+), Nd(3+), Sm(3+), and Eu(3+) in the low-ppb range (6-11 nM).

  15. Study on the violet LED-induced fluorescence spectra of thioredoxin reductase from human brain

    Institute of Scientific and Technical Information of China (English)

    Xiufeng Lan(兰秀风); Tao Yang(杨涛); Shumei Gao(高淑梅); Xiaosen Luo(骆晓森); Zhonghua Shen(沈中华); Jian Lu(陆建); Xiaowu Ni(倪晓武); Lin Xu(许琳)

    2003-01-01

    The technique of fluorescence spectroscopy is applied to study thioredoxin reductase (TrxR) in the ce;lsof human brain. Experimental results show that, by the violet light emitting diode (LED, λmax=407 nm)light irradiation, TrxR is able to emit three striking spectral bands (528 - 582 nm; 588 - 660 nm; 683 - 700nm). The fluorescence intensity is linear to the concentration of TrxR. The spectrum of denatured TrxR israther different from that of organized TrxR, which reflects the structure change between denatured TrxRand organized TrxR. Furthermore, physical and biochemical mechanisms of fluorescence production forLED light-induced TrxR spectra and its characteristics are analyzed. This paper may be useful to betterunderstand the structure of TrxR, and to provide new spectroscopic information to improve the resolutionfor this kind of biology structure.

  16. Ligand-induced evolution of intrinsic fluorescence and catalytic activity from cobalt ferrite nanoparticles.

    Science.gov (United States)

    Pal, Monalisa; Kundu, Anirban; Rakshit, Rupali; Mandal, Kalyan

    2015-06-08

    To develop CoFe(2)O(4) as magneto-fluorescent nanoparticles (NPs) for biomedical applications, it would be advantageous to identify any intrinsic fluorescence of this important magnetic material by simply adjusting the surface chemistry of the NPs themselves. Herein, we demonstrate that intrinsic multicolor fluorescence, covering the whole visible region, can be induced by facile functionalization of CoFe(2)O(4) NPs with Na-tartrate. Moreover, the functionalized CoFe(2)O(4) NPs also show unprecedented catalytic efficiency in the degradation of both biologically and environmentally harmful dyes, pioneering the potential application of these NPs in therapeutics and wastewater treatment. Detailed investigation through various spectroscopic tools unveils the story behind the emergence of this unique optical property of CoFe(2)O(4) NPs upon functionalization with tartrate ligands. We believe our developed multifunctional CoFe(2)O(4) NPs hold great promise for advanced biomedical and technological applications.

  17. Composition measurement of bicomponent droplets using laser-induced fluorescence of acetone

    Science.gov (United States)

    Maqua, C.; Depredurand, V.; Castanet, G.; Wolff, M.; Lemoine, F.

    2007-12-01

    Commercial fuels are complex mixtures, the evaporation of which remains particularly difficult to model. Experimental characterization of the differential vaporization of the components is a problem that is seldom addressed. In this paper, the evaporation of binary droplets made of ethyl-alcohol and acetone is investigated using a technique of measurement of the droplet composition developed in purpose. This technique exploits the laser induced fluorescence of acetone which acts as a fluorescent tracer as well as the more volatile component of the fuel associated with an accurate measurement of the droplet diameter by forward scattering interferometry. A model of the fluorescence intensity of the binary mixture, taking into account the absorption of the acetone molecules, is proposed and validated. The sensitivity of the technique is discussed. Finally, the reliability of the technique is demonstrated on binary combusting droplets in linear stream.

  18. Laser induced fluorescence spectroscopy of chemo-drugs as biocompatible fluorophores: irinotecan, gemcitabine and navelbine

    Science.gov (United States)

    Motlagh, N. S. Hosseini; Parvin, P.; Ghasemi, F.; Atyabi, F.; Jelvani, S.; Abolhosseini, S.

    2016-07-01

    The fluorescence nature of chemo-drugs is useful for simultaneous cancer diagnosis and therapy. Here, the laser induced fluorescence (LIF) properties of irinotecan, gemcitabine and navelbine are extensively investigated. The UV photons provoke the desired transitions of the several chemo-drugs by virtue of the XeCl laser at 308 nm. It is shown that LIF spectra are strongly dependent on the fluorophore concentration, while no spectral shift is measured for irinotecan, gemcitabine and navelbine because of a large Stokes shift. On the other hand, doxorubicin is characterized by a large overlapping between absorption and emission spectra giving rise to a sensible red shift. The fluorescence extinction α and self-quenching k coefficients as well as the quantum yield η f of those chemo-drugs are determined accordingly. In fact, irinotecan shows the highest quantum efficiency among the chemo-drugs of interest.

  19. A general method for the detection of large CAG repeat expansions by fluorescent PCR

    OpenAIRE

    Warner, J P; Barron, L H; Goudie, D; Kelly, K; Dow, D.; FitzPatrick, D.R.; Brock, D J

    1996-01-01

    The expansion of a tandemly repeated trinucleotide sequence, CAG, is the mutational mechanism for several human genetic diseases. We present a generally applicable PCR amplification method using a fluorescently labelled locus specific primer flanking the CAG repeat together with paired primers amplifying from multiple priming sites within the CAG repeat. Triplet repeat primed PCR (TP PCR) gives a characteristic ladder on the fluorescence trace enabling the rapid identification of large pathog...

  20. Carboxymethyl-β-cyclodextrin for Chiral Separation of Amino Acids Derivatized with Fluorescene-5-isothiocyanate by Capillary Electrophoresis and Laser-induced Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    Yu Yun CHEN; Wei WANG; Wei Ping YANG; Zhu Jun ZHANG

    2004-01-01

    A method using carboxymethyl-β-cyclodextrin(CM-β-CD) as selector for chiral separation of amino acids by capillary electrophoresis and laser-induced fluorescence detection was studied. Resolution was better than that obtained byβ-CD or HP-β-CD.

  1. Determination of Amino Acids in an Individual Erythrocyteby Capillary Electrophoresis with Intracellular FITC-derivatization and Laser-induced Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    Hua ZHANG; Wen Rui JIN

    2003-01-01

    A novel approach for analysis of amino acids in individual erythrocytes was established.In this method, the derivatization reagent was introduced into the living cells by electroporation.After derivatization, the amino acids in a single cell were determined by capillary electrophoresiswith laser-induced fluorescence detection.

  2. Determination of torasemide by fluorescence quenching method with some dihalogenated fluorescein dyes as probes

    Science.gov (United States)

    Cui, Zhiping; Liu, Shaopu; Liu, Zhongfang; Li, Yuanfang; Hu, Xiaoli; Tian, Jing

    2013-10-01

    A novel fluorescence quenching method for the determination of torasemide (TOR) with some dihalogenated fluorescein dyes as fluorescence probes was developed. In acidulous medium, TOR could interact with some dihalogenated fluorescein dyes such as dichlorofluorescein (DCF), dibromofluorescein (DBF) and diiodofluorescein (DIF) to form binary complexes, which could lead to fluorescence quenching of above dihalogenated fluorescein dyes. The maximum fluorescence emission wavelengths were located at 532 nm (TOR-DCF), 535 nm (TOR-DBF) and 554 nm (TOR-DIF). The relative fluorescence intensities (ΔF = F0 - F) were proportional to the concentration of TOR in certain ranges. The detection limits were 4.8 ng mL-1 for TOR-DCF system, 9.8 ng mL-1 for TOR-DBF system and 35.1 ng mL-1 for TOR-DIF system. The optimum reaction conditions, influencing factors were studied; and the effect of coexisting substances was investigated owing to the highest sensitivity of TOR-DCF system. In addition, the reaction mechanism, composition and structure of the complex were discussed by quantum chemical calculation and Job's method. The fluorescence quenching of dihalogenated fluorescein dyes by TOR was a static quenching process judging from the effect of temperature and the Stern-Volmer plots. The method was satisfactorily applied to the determination of TOR in tablets and human urine samples.

  3. Dexamethasone-Inducible Green Fluorescent Protein Gene Expression in Transgenic Plant Cells

    Institute of Scientific and Technical Information of China (English)

    Wei Tang; Hilary Collver; Katherine Kinken

    2004-01-01

    Genomic research has made a large number of sequences of novel genes or expressed sequence tags available. To investigate functions of these genes, a system for conditional control of gene expression would be a useful tool. Inducible transgene expression that uses green fluorescent protein gene (gfp) as a reporter gene has been investigated in transgenic cell lines of cotton (COT; Gossypium hirsutum L.), Fraser fir [FRA; Abies fraseri (Pursh) Poir], Nordmann fir (NOR; Abies nordmanniana Lk.), and rice (RIC; Oryza sativa L. Cv. Radon). Transgenic cell lines were used to test the function of the chemical inducer dexamethasone. Inducible transgene expression was observed with fluorescence and confocal microscopy, and was confirmed by northern blot analyses. Dexamethasone at 5 mg/L induced gfp expression to the nearly highest level 48 h after treatment in COT, FRA, NOR, and RIC. Dexamethasone at 10 mg/L inhibited the growth of transgenic cells in FRA and NOR, but not COT and RIC. These results demonstrated that concentrations of inducer for optimum inducible gene expression system varied among transgenic cell lines. The inducible gene expression system described here was very effective and could be valuable in evaluating the function of novel gene.

  4. Simultaneous measurements of temperature, density, and pressure in a supersonic turbulent flow using laser-induced fluorescence

    Science.gov (United States)

    Mckenzie, R. L.; Gross, K. P.; Logan, P.

    1985-01-01

    A pulsed laser-induced fluorescence technique is described that provides simultaneous measurements of temperature, density, and pressure in low-temperature, turbulent flows. The measurements are made with spatial and temporal resolution comparable to that obtained with modern laser anemometer techniques used for turbulent boundary layer research. The capabilities of the method are briefly described and its demonstration in a simple two-dimensional turbulent boundary layer at Mach 2 is reported. The results are compared with conventional hot-wire anemometer data obtained in the same flow.

  5. [Two-photon excitation fluorescence of 5-ALA induced PpIX in DHL cells].

    Science.gov (United States)

    Huang, Zu-Fang; Chen, Rong; Li, Yong-Zeng; Chen, Guan-Nan; Chen, Xian-Ling; Feng, Shang-Yuan; Jia, Pei-Min

    2008-11-01

    Two-photon fluorescence microscopy is a novel imaging technique, which is primarily sensitive to a specimen's response coming from an in-focus plane, thus has low photo-bleaching and photo-damage to biological samples. 5-ALA induced production of PpIX in DHL cells was excited by 820 nm femtosecond laser; two-photon excitation fluorescence of single cell was obtained in Lambda mode of laser scanning confocal microscope. The specific fluorescence intensity of PpIX which accumulated in DHL cells was measured at 2, 4 and 10 mmol x L(-1) concentration of 5-ALA with different incubation time, which reflected the kinetics of 5-ALA accumulated in DHL cells. Accumulation of PpIX in DHL cells was a dynamic change process. Biphasic alterations of PpIX accumulation were noted: PpIX content enhanced with the increasing time and reached the maximal value around 3 h, however PpIX content decreased in the subsequent incubation time. Results indicate that two-photon fluorescence based on laser scanning microscope can be a useful technology for studying the kinetics of 5-ALA induced PpIX production in DHL cells and other leukemia cells.

  6. Aggregation-induced emissive nanoparticles for fluorescence signaling in a low cost paper-based immunoassay.

    Science.gov (United States)

    Engels, Jan F; Roose, Jesse; Zhai, Demi Shuang; Yip, Ka Man; Lee, Mei Suet; Tang, Ben Zhong; Renneberg, Reinhard

    2016-07-01

    Low cost paper based immunoassays are receiving interest due to their fast performance and small amounts of biomolecules needed for developing an immunoassay complex. In this work aggregation-induced emissive (AIE) nanoparticles, obtained from a diastereoisomeric mixture of 1,2-di-(4-hydroxyphenyl)-1,2-diphenylethene (TPEDH) in a one-step top-down method, are characterized through Dynamic Light Scattering (DLS), Scanning Electron Microscopy (SEM), and Zeta potential. By measuring the Zeta potential before and after labeling the nanoparticles with antibodies we demonstrate that the colloidal system is stable in a wide pH-range. The AIE-active nanoparticles are deposited on chitosan and glutaraldehyde modified paper pads overcoming the common aggregation-caused quenching (ACQ) effect. Analyte concentrations from 1000ng and below are applied in a model immunocomplex using Goat anti-Rabbit IgG and Rabbit IgG. In the range of 7.81ng-250ng, linear trends with a high R(2) are observed, which leads to a strong increase of the blue fluorescence from the TPEDH nanoparticles.

  7. Restriction Enzyme Pattern Analysis of Mycobacteria DNA by Capillary Electrophoresis with Laser-induced Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    Li Yuanqian; Wang Guoqing; Mi Jianping; Zhou Ying; Zeng Hongyan; Zhang Chaowu

    2006-01-01

    A new method for rapidly detecting restriction enzyme patterns of Mycobacterium DNA using capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD)was developed.Polymerase chain reaction was used to amplify a 439-bp fragment of a 65,000-kDa(Mr)heat shock protein gene(hsp65)of Mycobacterium.After digesting amplification products by BstEII and HaeIII,patterns of enzyme cleavage products were detected by both CE-LIFD and agarose gel electrophoresis(AGE),respectively.Experimental parameters of CE were optimized.Restriction enzyme patterns of Mycobacterium DNA were detected in optimum electrophoresis conditions:a coated capillary column with a length of 50 cm and an internal diameter of 100 μm,an electrophoresis buffer of 45 mmol/1 Tris-boric acid-ethylenediaminetetraacetic acid,and a running voltage of 11 kV.The restriction enzyme patterns for eight species of mycobacteria were studied.Relative standard deviations of the relative migration times of DNA segments were<3.6%.Compared with AGE,CE is more outstanding in resolution and detection time,and it can be applied as a more effective means to DNA restriction enzyme pattern analysis.

  8. Modelling satellite-level solar-induced chlorophyll fluorescence and its comparison with OCO-2 observations

    Science.gov (United States)

    Pradhan, Rohit; Gohel, Ankit

    2016-04-01

    Solar Induced chlorophyll Fluorescence (SIF) is a direct measure of photosynthesis rate as it is emitted by the photosynthetic system. This paper reports a method to model SIF over India by assimilating spatial inputs (LAI, Chlorophyll content etc.) into FluorMOD leaf and canopy model. Modelled SIF was then compared to Orbiting Carbon Observatory (OCO-2) SIF observations from September 2014 to August 2015. Modelled SIF at 757 nm averaged over the country varied between 0.18 to 0.33 W m-2 sr-1 μm-1 whereas SIF at 771 nm varied between 0.10 to 0.19 W m-2 sr-1 μm-1. OCO-2 observed SIF at 757 nm averaged over the country ranged from 0.18 to 0.42 Wm-2sr-1μm-1. Fairly good agreement (r=0.77, p<0.01 at 757nm; r=0.71, p<0.05 at 771 nm) was observed between modelled and observed SIF except for summer months of April and May. This paper presents a new approach to upscale a leaf and canopy level model to estimate SIF over entire country.

  9. Assessment of Parturition with Cervical Light-Induced Fluorescence and Uterine Electromyography

    Directory of Open Access Journals (Sweden)

    Miha Lucovnik

    2013-01-01

    Full Text Available Parturition involves increasing compliance (ripening of the uterine cervix and activation of the myometrium. These processes take place in a different time frame. Softening and shortening of the cervix starts in midpregnancy, while myometrial activation occurs relatively close to delivery. Methods currently available to clinicians to assess cervical and myometrial changes are subjective and inaccurate, which often causes misjudgments with potentially adverse consequences. The inability to reliably diagnose true preterm labor leads to unnecessary treatments, missed opportunities to improve neonatal outcome, and inherently biased research of treatments. At term, the likelihood of cesarean delivery depends on labor management, which in turn depends on accurate assessments of cervical change and myometrial contractility. Studies from our group and others show that noninvasive measurements of light-induced fluorescence (LIF of cervical collagen and uterine electromyography (EMG objectively detect changes in the composition of the cervix and myometrial preparedness to labor and are more reliable than clinical observations alone. We present a conceptual model of parturition constructed on cervical LIF and uterine EMG studies. We also explore how these methodologies could be helpful with managing patients experiencing preterm contractions and with optimizing labor management protocols aimed to reduce cesarean section.

  10. Fluorescent nanodiamond tracking reveals intraneuronal transport abnormalities induced by brain-disease-related genetic risk factors

    Science.gov (United States)

    Haziza, Simon; Mohan, Nitin; Loe-Mie, Yann; Lepagnol-Bestel, Aude-Marie; Massou, Sophie; Adam, Marie-Pierre; Le, Xuan Loc; Viard, Julia; Plancon, Christine; Daudin, Rachel; Koebel, Pascale; Dorard, Emilie; Rose, Christiane; Hsieh, Feng-Jen; Wu, Chih-Che; Potier, Brigitte; Herault, Yann; Sala, Carlo; Corvin, Aiden; Allinquant, Bernadette; Chang, Huan-Cheng; Treussart, François; Simonneau, Michel

    2017-05-01

    Brain diseases such as autism and Alzheimer's disease (each inflicting >1% of the world population) involve a large network of genes displaying subtle changes in their expression. Abnormalities in intraneuronal transport have been linked to genetic risk factors found in patients, suggesting the relevance of measuring this key biological process. However, current techniques are not sensitive enough to detect minor abnormalities. Here we report a sensitive method to measure the changes in intraneuronal transport induced by brain-disease-related genetic risk factors using fluorescent nanodiamonds (FNDs). We show that the high brightness, photostability and absence of cytotoxicity allow FNDs to be tracked inside the branches of dissociated neurons with a spatial resolution of 12 nm and a temporal resolution of 50 ms. As proof of principle, we applied the FND tracking assay on two transgenic mouse lines that mimic the slight changes in protein concentration (∼30%) found in the brains of patients. In both cases, we show that the FND assay is sufficiently sensitive to detect these changes.

  11. Intercomparison of Hantzsch and fiber-laser-induced-fluorescence formaldehyde measurements

    Science.gov (United States)

    Kaiser, J.; Li, X.; Tillmann, R.; Acir, I.; Holland, F.; Rohrer, F.; Wegener, R.; Keutsch, F. N.

    2014-06-01

    Two gas-phase formaldehyde (HCHO) measurement techniques, a modified commercial wet-chemical instrument based on Hantzsch fluorimetry and a custom-built instrument based on fiber laser-induced fluorescence (FILIF), were deployed at the atmospheric simulation chamber SAPHIR (Simulation of Atmospheric PHotochemistry In a large Reaction Chamber) to compare the instruments' performances under a range of conditions. Thermolysis of para-HCHO and ozonolysis of 1-butene were used as HCHO sources, allowing for calculations of theoretical HCHO mixing ratios. Calculated HCHO mixing ratios are compared to measurements, and the two measurements are also compared. Experiments were repeated under dry and humid conditions (RH 60%) to investigate the possibility of a water artifact in the FILIF measurements. The ozonolysis of 1-butene also allowed for the investigation of an ozone artifact seen in some Hantzsch measurements in previous intercomparisons. Results show that under all conditions the two techniques are well correlated (R2 ≥ 0.997), and linear regression statistics show measurements agree with within stated uncertainty (15% FILIF + 5% Hantzsch). No water or ozone artifacts are identified. While a slight curvature is observed in some Hantzsch vs. FILIF regressions, the potential for variable instrument sensitivity cannot be attributed to a single instrument at this time. Measurements at low concentrations highlight the need for a secondary method for testing the purity of air used in instrument zeroing and the need for further FILIF White cell outgassing experiments.

  12. The effects of illumination and focal distance on light-induced fluorescence images in vitro.

    Science.gov (United States)

    Pretty, I A; Ellwood, P G; Davies, R M; Worthington, H W; Ellwood, R P

    2006-01-01

    When using quantitative light-induced fluorescence a number of factors can influence illumination level. The purpose of this study was to investigate, using a high-resolution camera and fibre-optic light source, the impact of illumination level and focal distance on common quantitative light-induced fluorescence outcomes. Twenty-four extracted teeth were examined using 6 illumination levels and 4 focal distances. Analysis was conducted using multiple linear regression models fitted to log DeltaQ, log DeltaF and log area with clustering of teeth and robust standard errors. Separate models were used for the different light and focal levels. The regression coefficients were significant for both DeltaQ and DeltaF but not area. Despite the significant regressions the actual effect was very small, and unlikely to confound clinical trial or practice results.

  13. Parameter estimation method for blurred cell images from fluorescence microscope

    Science.gov (United States)

    He, Fuyun; Zhang, Zhisheng; Luo, Xiaoshu; Zhao, Shulin

    2016-10-01

    Microscopic cell image analysis is indispensable to cell biology. Images of cells can easily degrade due to optical diffraction or focus shift, as this results in low signal-to-noise ratio (SNR) and poor image quality, hence affecting the accuracy of cell analysis and identification. For a quantitative analysis of cell images, restoring blurred images to improve the SNR is the first step. A parameter estimation method for defocused microscopic cell images based on the power law properties of the power spectrum of cell images is proposed. The circular radon transform (CRT) is used to identify the zero-mode of the power spectrum. The parameter of the CRT curve is initially estimated by an improved differential evolution algorithm. Following this, the parameters are optimized through the gradient descent method. Using synthetic experiments, it was confirmed that the proposed method effectively increased the peak SNR (PSNR) of the recovered images with high accuracy. Furthermore, experimental results involving actual microscopic cell images verified that the superiority of the proposed parameter estimation method for blurred microscopic cell images other method in terms of qualitative visual sense as well as quantitative gradient and PSNR.

  14. Discrimination of corn from monocotyledonous weeds with ultraviolet (UV) induced fluorescence.

    Science.gov (United States)

    Panneton, Bernard; Guillaume, Serge; Samson, Guy; Roger, Jean-Michel

    2011-01-01

    In production agriculture, savings in herbicides can be achieved if weeds can be discriminated from crop, allowing the targeting of weed control to weed-infested areas only. Previous studies demonstrated the potential of ultraviolet (UV) induced fluorescence to discriminate corn from weeds and recently, robust models have been obtained for the discrimination between monocots (including corn) and dicots. Here, we developed a new approach to achieve robust discrimination of monocot weeds from corn. To this end, four corn hybrids (Elite 60T05, Monsanto DKC 26-78, Pioneer 39Y85 (RR), and Syngenta N2555 (Bt, LL)) and four monocot weeds (Digitaria ischaemum (Schreb.) I, Echinochloa crus-galli (L.) Beauv., Panicum capillare (L.), and Setaria glauca (L.) Beauv.) were grown either in a greenhouse or in a growth cabinet and UV (327 nm) induced fluorescence spectra (400 to 755 nm) were measured under controlled or uncontrolled ambient light intensity and temperature. This resulted in three contrasting data sets suitable for testing the robustness of discrimination models. In the blue-green region (400 to 550 nm), the shape of the spectra did not contain any useful information for discrimination. Therefore, the integral of the blue-green region (415 to 455 nm) was used as a normalizing factor for the red fluorescence intensity (670 to 755 nm). The shape of the normalized red fluorescence spectra did not contribute to the discrimination and in the end, only the integral of the normalized red fluorescence intensity was left as a single discriminant variable. Applying a threshold on this variable minimizing the classification error resulted in calibration errors ranging from 14.2% to 15.8%, but this threshold varied largely between data sets. Therefore, to achieve robustness, a model calibration scheme was developed based on the collection of a calibration data set from 75 corn plants. From this set, a new threshold can be estimated as the 85% quantile on the cumulative frequency

  15. Clinical diagnosis of fissure caries with conventional and laser-induced fluorescence techniques

    OpenAIRE

    Chu, C. H.; Lo, E. C. M.; You, D. S. H.

    2010-01-01

    We studied the in vivo validity of dentinal fissure caries diagnosis by visual examination, bitewing radiography, and use of a laser-induced fluorescence device (DIAGNOdent). A total of 144 and second molars with macroscopically intact occlusal surfaces in 41 Chinese young adults were examined visually, by bitewing radiography, and by DIAGNOdent. Visual examination after pit and fissure opening was used as the reference standard. The sensitivity and specificity of detecting caries that had ex...

  16. Study on Fluorescence Analytical Method of Micro Th in Pu

    Institute of Scientific and Technical Information of China (English)

    QIAN; Hong-juan; ZHANG; Li-hua; LIU; Huan-liang; FAN; De-jun

    2013-01-01

    As one of impurities,Th is needed to be determined for control quality of Pu product.Due to the high radioactivity and fatal toxicity of Pu,it is very difficult to measure the micro Th in Pu product.It is necessary to develop a sensitive method for determining the micro Th in Pu under the described harsh condition.

  17. A restaining method to restore fluorescence in faded preparations of tissues treated with the indirect immunofluorescence technique.

    Science.gov (United States)

    Weinstein, W M; Lechago, J

    1977-01-01

    We report a restainin method for restoring fluorescence in paraffin-embedded tissue sections previously treated with the indirect immunofluorescence technique. Antisera to gastrin and group II pepsinogens were used. Fluorescence was restored in completely faded sections retrived from storage files, as well as in sections that had faded partially either with exposure to fluorescence microscope illumination or after counterstaining with hematoxylin and eosin.

  18. Comparison of Fluorescence Microscopy and Different Growth Media Culture Methods for Acanthamoeba Keratitis Diagnosis.

    Science.gov (United States)

    Peretz, Avi; Geffen, Yuval; Socea, Soergiu D; Pastukh, Nina; Graffi, Shmuel

    2015-08-01

    Acanthamoeba keratitis (AK), a potentially blinding infection of the cornea, is caused by a free-living protozoan. Culture and microscopic examination of corneal scraping tissue material is the conventional method for identifying Acanthamoeba. In this article, we compared several methods for AK diagnosis of 32 patients: microscopic examination using fluorescent dye, specific culture on growth media-non-nutrient agar (NNA), culture on liquid growth media-peptone yeast glucose (PYG), and TYI-S-33. AK was found in 14 patients. Thirteen of the specimens were found AK positive by fluorescence microscopic examination, 11 specimens were found AK positive on PYG growth media, and 9 specimens were found AK positive on TYI-S-33 growth media. Only five specimens were found AK positive on NNA growth media. Therefore, we recommend using fluorescence microscopy technique and culture method, especially PYG liquid media.

  19. Methyl-β-Cyclodextrin /Cetyltrimethyl Ammonium Bromide Synergistic Sensitized Fluorescence Method for the Determination of Levofloxacin.

    Science.gov (United States)

    Ren, Qiuyi; Zhu, Xiashi

    2016-03-01

    A novel method of methyl-β-cyclodextrin (methyl-β-CD) and cetyltrimethyl ammonium bromide (CTAB) synergistic sensitized fluorescence analysis to determine levofloxacin (LVFX) was developed. The results were shown that the fluorescence intensity of LVFX was increased a lot in the system of methyl-β-cyclodextrin-CTAB medium. Under the conditions of λ(ex/em )= 330/507 nm and pH 4.5, the linear range and the detection limit for LVFX were found to be 0.040 ~ 4.0 μg/mL and 0.3 ng/mL, respectively. The mechanism of sensitized fluorescence method was discussed by the solubilization capacity and the microenvironment of medium. The proposed method has been applied for the determination of LVFX in eye drops real samples and human serum with satisfactory recovery.

  20. Detection of the onset of glyphosate-induced soybean plant injury through chlorophyll fluorescence signal extraction and measurement

    Science.gov (United States)

    Zhao, Feng; Guo, Yiqing; Huang, Yanbo; Reddy, Krishna N.; Zhao, Yanhua; Molin, William T.

    2015-01-01

    In this study, chlorophyll fluorescence (ChlF) was used to detect the onset of soybean plant injury from treatment of glyphosate, the most widely used herbicide. Thirty-six pots of nonglyphosate-resistant soybean were randomly divided into three groups and treated with different doses of glyphosate solutions. The three treatment groups were control (CTRL) group (with no glyphosate treatment), 0.25X group (treated with 0.217 kg.ae/ha solution of glyphosate), and 0.5X group (treated with 0.433 kg.ae/ha solution of glyphosate). Three kinds of fluorescence measurements, steady-state fluorescence spectra, Kautsky effect parameters, and ChlF-related spectral indices were extracted and generated from the measurements in the glyphosate treatment experiment. The mean values of these fluorescence measurements for each of the CTRL group, the 0.25X group, and the 0.5X group were calculated. Glyphosate-induced leaf injury was then analyzed by examining the separability of these mean values at 6, 24, 48, and 72 hours after the treatment (HAT). Results indicate that the peak position of far-red ChlF shows an obvious blue shift for glyphosate-treated soybean, and peak values of steady-state fluorescence spectra for the three groups can be significantly distinguished from each other at 48 HAT and later. Four Kautsky effect parameters, Fv, Fv/Fm, Area, and PI, are parameters sensitive to glyphosate treatment, showing some differences between the CTRL group and treated groups at 24 HAT, and significant differences among the three groups at and beyond 48 HAT. Moreover, ChlF-related spectral indices, R6832/(R675.R690) and R690/R655, are also shown to be useful in detection of the glyphosate injury, though they are less effective than the steady-state fluorescence spectra and the Kautsky effect parameters. Based on the presented results, it can be concluded that glyphosate-induced soybean injury can be detected in a timely manner by the ChlF measurements, and this method has the

  1. Diagnostic efficacy of Ziehl-Neelsen method against fluorescent microscopy in detection of acid fast bacilli

    Institute of Scientific and Technical Information of China (English)

    Soham Gupta; Vishnu Prasad Shenoy; Indira Bairy; MuralidharanS

    2010-01-01

    Objective:To investigate the application of Ziehl-Neelsen (Z-N) and fluorescent microscopy in detection of acid fast bacilli (AFB).Methods: Duplicate smears were prepared from 260 sputum samples and stained with Z-N and fluorescent staining (FS) methods. The efficiency of both methods in primary diagnosis of tuberculosis were evaluated.Results:The smears were positive for AFB in 15 (5.77%) samples by Z-N staining method and in 16 (6.15%) samples by FS method. The sensitivity and specificity of Z-N staining method against FS method were 93.75% and 100% respectively.Conclusions: Though lesser cost-effective than Z-N, FS method is a more sensitive and better case finding tool in detection of AFB.

  2. Modeling the impact of spectral sensor configurations on the FLD retrieval accuracy of sun-induced chlorophyll fluorescence

    NARCIS (Netherlands)

    Damm, A.; Erler, A.; Hillen, W.; Meroni, M.; Schaepman, M.E.; Verhoef, W.; Rascher, U.

    2011-01-01

    Chlorophyll fluorescence is related to photosynthesis and can serve as a remote sensing proxy for estimating photosynthetic energy conversion and carbon uptake. Recent advances in sensor technology allow remote measurements of the sun-induced chlorophyll fluorescence signal (Fs) at leaf and canopy s

  3. Violet diode laser-induced chlorophyll fluorescence: a tool for assessing mosaic disease severity in cassava (Manihot esculenta Crantz) cultivars.

    Science.gov (United States)

    Anderson, Benjamin; Eghan, Moses J; Asare-Bediako, Elvis; Buah-Bassuah, Paul K

    2012-01-01

    Violet diode laser-induced chlorophyll fluorescence was used in agronomical assessment (disease severity and average yield per plant). Because cassava (Manihot esculenta Crantz) is of economic importance, improved cultivars with various levels of affinity for cassava mosaic disease were investigated. Fluorescence data correlated with cassava mosaic disease severity levels and with the average yield per plant.

  4. Microalgae amino acid extraction and analysis at nanomolar level using electroporation and capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Nehmé, Reine; Atieh, Carla; Fayad, Syntia; Claude, Bérengère; Chartier, Agnès; Tannoury, Mona; Elleuch, Fatma; Abdelkafi, Slim; Pichon, Chantal; Morin, Philippe

    2017-01-01

    Amino acids play a key role in food analysis, clinical diagnostics, and biochemical research. Capillary electrophoresis with laser-induced fluorescence detection was used for the analysis of several amino acids. Amino acid labeling with fluorescein isothiocyanate was conducted using microwave-assisted derivatization at 80°C (680 W) during only 150 s. Good electrophoretic resolution was obtained using a background electrolyte composed of sodium tetraborate buffer (100 mM; pH 9.4) and β-cyclodextrin (10 mM), and the limits of quantification were 3-30 nM. The developed capillary electrophoresis with laser-induced fluorescence method was used to analyze amino acids in Dunaliella salina green algae grown under different conditions. A simple extraction technique based on electroporation of the cell membrane was introduced. A home-made apparatus allowed the application of direct and alternating voltages across the electrochemical compartment containing a suspension of microalgae in distilled water at 2.5 g/L. A direct voltage of 12 V applied for 4 min gave the optimum extraction yield. Results were comparable to those obtained with accelerated-solvent extraction. The efficiency of electroporation in destroying microalgae membranes was shown by examining the algae surface morphology using scanning electron microscopy. Stress conditions were found to induce the production of amino acids in Dunaliella salina cells. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

  5. Correlation between the In Vitro Functionality of Stored Platelets and the Cytosolic Esterase-Induced Fluorescence Intensity with CMFDA.

    Science.gov (United States)

    Wang, Jiexi; Yi, Xiaoyang; Liu, Minxia; Zhou, Qian; Ren, Suping; Wang, Yan; Yang, Chao; Zhou, Jianwei; Han, Ying

    2015-01-01

    It has been hypothesized that the cytosolic esterase-induced fluorescence intensity (CEIFI) from carboxy dimethyl fluorescein diacetate (CMFDA) in platelets may related to platelet functions. In the present study, we measured the change of CEIFI in platelets during storage, and examined the correlations of CEIFI with the in vitro functionality of stored platelets, including the ADP-induced aggregation activity, hypotonic shock response, expression of CD62P as well as platelet apoptosis. The CEIFI of fresh platelets, when tested at 10 μM CMFDA, the mean fluorescence intensity index (MFI) was 305.9 ± 49.9 (N = 80). After 1-day storage, it was 203.8 ± 34.4, the CEIFI of the stored platelets started to decline significantly, and reduced to 112.7 ±27.7 after 7-day storage. The change in CEIFI is highly correlated to all four functional parameters measured, with the correlation coefficients being 0.9813, 0.9848, -0.9945 and -0.9847 for the ADP-induced aggregation activity, hypotonic shock response (HSR), expression of CD62P and platelet apoptosis respectively. The above results show that the CEIFI measurement of platelets represents well the viability and functional state of in vitro stored platelets. This may be used as a convenient new method for quality evaluation for stored platelets if this result can be further validated by the following clinical trials.

  6. A Fluorescence-Based Method for Rapid and Direct Determination of Polybrominated Diphenyl Ethers in Water

    Directory of Open Access Journals (Sweden)

    Huimei Shan

    2015-01-01

    Full Text Available A new method was developed for rapid and direct measurement of polybrominated diphenyl ethers (PBDEs in aqueous samples using fluorescence spectroscopy. The fluorescence spectra of tri- to deca-BDE (BDE 28, 47, 99, 153, 190, and 209 commonly found in environment were measured at variable emission and excitation wavelengths. The results revealed that the PBDEs have distinct fluorescence spectral profiles and peak positions that can be exploited to identify these species and determine their concentrations in aqueous solutions. The detection limits as determined in deionized water spiked with PBDEs are 1.71–5.82 ng/L for BDE 28, BDE 47, BDE 190, and BDE 209 and 45.55–69.95 ng/L for BDE 99 and BDE 153. The effects of environmental variables including pH, humic substance, and groundwater chemical composition on PBDEs measurements were also investigated. These environmental variables affected fluorescence intensity, but their effect can be corrected through linear additivity and separation of spectral signal contribution. Compared with conventional GC-based analytical methods, the fluorescence spectroscopy method is more efficient as it only uses a small amount of samples (2–4 mL, avoids lengthy complicated concentration and extraction steps, and has a low detection limit of a few ng/L.

  7. X-RAY FLUORESCENCE ANALYSIS OF HANFORD LOW ACTIVITY WASTE SIMULANTS METHOD DEVELOPMENT

    Energy Technology Data Exchange (ETDEWEB)

    Jurgensen, A; David Missimer, D; Ronny Rutherford, R

    2007-08-08

    The x-ray fluorescence laboratory (XRF) in the Analytical Development Directorate (ADD) of the Savannah River National Laboratory (SRNL) was requested to develop an x-ray fluorescence spectrometry method for elemental characterization of the Hanford Tank Waste Treatment and Immobilization Plant (WTP) pretreated low activity waste (LAW) stream to the LAW Vitrification Plant. The WTP is evaluating the potential for using XRF as a rapid turnaround technique to support LAW product compliance and glass former batching. The overall objective of this task was to develop an XRF analytical method that provides rapid turnaround time (<8 hours), while providing sufficient accuracy and precision to determine variations in waste.

  8. Comparison between the indocyanine green fluorescence and blue dye methods for sentinel lymph node biopsy using novel fluorescence image-guided resection equipment in different types of hospitals.

    Science.gov (United States)

    He, Kunshan; Chi, Chongwei; Kou, Deqiang; Huang, Wenhe; Wu, Jundong; Wang, Yabing; He, Lifang; Ye, Jinzuo; Mao, Yamin; Zhang, Guo-Jun; Wang, Jiandong; Tian, Jie

    2016-12-01

    Sentinel lymph node biopsy (SLNB) has become a standard of care to detect axillary lymph metastasis in early-stage breast cancer patients with clinically negative axillary lymph nodes. Current SLNB detection modalities comprising a blue dye, a radioactive tracer, or a combination of both have advantages as well as disadvantages. Thus, near-infrared fluorescence imaging using indocyanine green (ICG) has recently been regarded as a novel method that has generated interest for SLNB around the world. However, the lack of appropriate fluorescence imaging systems has hindered further research and wide application of this method. Therefore, we developed novel fluorescence image-guided resection equipment (FIRE) to detect sentinel lymph nodes (SLNs). Moreover, to compare the ICG fluorescence imaging method with the blue dye method and to explore the universal feasibility of the former, a different type of hospital study was conducted. Ninety-nine eligible patients participated in the study at 3 different types of hospitals. After subcutaneous ICG allergy testing, all the patients were subcutaneously injected with methylene blue and ICG into the subareolar area. Consequently, 276 SLNs (range 1-7) were identified in 98 subjects (detection rate: 99%) by using the ICG fluorescence imaging method. In contrast, the blue dye method only identified 202 SLNs (range 1-7) in 91 subjects (detection rate: 91.92%). Besides, the results of the fluorescence imaging method were similar in the 3 hospitals. Our findings indicate the universal feasibility of the ICG fluorescence imaging method for SLNB using the fluorescence image-guided resection equipment in early breast cancer detection. Copyright © 2016 Elsevier Inc. All rights reserved.

  9. Simultaneous detection of ultraviolet B-induced DNA damage using capillary electrophoresis with laser-induced fluorescence

    Energy Technology Data Exchange (ETDEWEB)

    Guthrie, Jeffrey W., E-mail: jeff.guthrie@emich.edu; Limmer, Robert T.; Brooks, Eric A.; Wisnewski, Chelsea C.; Loggins-Davis, Nnekia D.; Bouzid, Abderraouf

    2015-01-01

    Highlights: • CE–LIF was developed for simultaneous detection of UV-induced DNA photoproducts. • Fluorescent quantum dot reporters enabled detection of small amounts of photoproducts. • Photoproducts were detected after 65 J m{sup −2} of fluence from a UVB lamp in ∼6 ng of DNA. • Natural sunlight induced cyclobutane pyrimidine dimers after only 15 min of exposure. - Abstract: An immunoassay based on CE–LIF was developed for the simultaneous detection of cyclobutane pyrimidine dimers (CPDs) and pyrimidine 6-4 pyrimidone photoproducts (6-4PPs) in genomic DNA irradiated with UVB or natural sunlight. Human cells were first exposed to varying amounts of UVB or natural sunlight to induce DNA damage. Genomic DNA was extracted and incubated with anti-CPD and anti-6-4PP primary antibodies attached to secondary antibodies with a fluorescent quantum dot (QD) reporter that emitted either red or yellow fluorescence. CE was used to separate the unbound antibodies from those bound to the photoproducts, and LIF with appropriate optical filters was used to separate the fluorescence signals from each QD to individual photomultiplier tubes for simultaneous photoproduct detection. Using this strategy, photoproducts were detected from ∼6 ng (200 ng μL{sup −1}) of DNA under a low UVB fluence of 65 J m{sup −2} for CPDs or 195 J m{sup −2} for 6-4PPs. This assay was also the first to demonstrate the detection of CPDs in human cells after only 15 min of irradiation under natural sunlight.

  10. Laser-induced Native Fluorescence Detection of Organic Molecules in Hydrothermal Vent Rocks

    Science.gov (United States)

    Harju, E.; Kidd, R. D.; Bhartia, R.; Conrad, P. G.

    2004-12-01

    We have developed a Multi-channel Deep Ultraviolet Excitation (McDuve) fluorescence detector that has been deployed at several Pacific hydrothermal vent sites [1]. The in situ McDuve detector was able to detect organic molecules at the vent site on rock surfaces and in the water, the signatures being distinguishable one from the other. The McDuve fluorescence detector uses a 224.3 nm helium-silver hollow cathode laser to induce native fluorescence from a sample. Spectral separation is achieved with optical band-pass filters which are coupled to photomultiplier tubes (PMTs) for detection. Samples were recovered at the vent sites and returned from the expedition for bench-top analysis for correlation of the McDuve observations with standard analytical tools-GCMS and X-ray diffraction (for mineralogical ID), as well as with a bench-top version of the McDuve fluorescence detector. Here we report the corroborative results of the laboratory studies. Several preserved samples were subjected to 224.3 nm ultraviolet excitation under wet and dry conditions. Organic molecules were detected on the wet samples analyzed in the lab, corroborating the in situ McDuve data. The fluorescence emission wavelengths associated with the detected organic molecules suggest they are 3-5 ring polycyclic aromatic hydrocarbons [2,3]. The samples were also pyrolized at 500 ºC to decompose any organic molecules present and subsequently reanalyzed. This McDuve analysis revealed a significant decrease in laser induced native fluorescence, a result consistent with the pyrolytic decomposition of the organic content of the rock samples. [1] Conrad, P.G., A.L. Lane, R. Bhartia, W. Hug, (March 2004) Optical Detection of Organic Chemical Biosignatures at Hydrothermal Vents 35th Lunar Plan. Sci. XXXV, 2055. [2] Karcher, W. (1985), Spectral Atlas of Polycyclic Aromatic Compounds, vol. I, Kluwer Academic Publishing Company, Dordrecht, Holland. [3] Bhartia, R., McDonald, G.D., Salas, E.C., Hug, W., Reid, R

  11. Laser ablation laser induced fluorescence for sensitive detection of heavy metals in water

    Science.gov (United States)

    Godwal, Yogesh

    Laser Induced Breakdown Spectroscopy LIBS is a fast non-contact technique for the analysis of the elemental composition using spectral information of the emission from a laser-induced plasma. For the LIBS studies in this thesis the focus has been in using very low energy, microjoule pulses in order to give high spatial resolution and minimize the laser system requirements. This is a regime that we refer to as microLIBS. Under such conditions it is important to maximize the signal detected to give the lowest limit of detection LOD possible. One technique to improve the signal to noise ratios is by coupling LIBS with Laser Induced Fluorescence. This is a technique where the first pulse creates a vapor plume and the second pulse tuned to a resonant absorption line of the species of interest re-excites the plume. We term this technique as Laser ablation Laser Induced Fluorescence LA-LIF. We have been investigating the performance of LA-LIF at low pulse energies (≤ 1 mJ for both pulses) for the detection of elemental contaminants in water. This technique allows reasonable performance compared to high energy single-pulse LIBS, but at a much reduced total energy expenditure. This allows LODs in the parts per billion range ppb range which typically cannot be obtained with low energy single pulse probing of the systems. This approach or exceeds the sensitivities which can be obtained with many shots using much larger energy systems. In this thesis we investigated the performance of LIBS at low pulse energies for the detection of Pb as a contaminant in water. An LOD of 70 ppb was obtained for an accumulation of 100 shots with the ablation laser pulse energy of 250 muJ and an excitation laser pulse energy of 8 muJ. A systematic study of the detector conditions was made for the system for the detection of Pb. Scaling laws for the LOD in terms of the pump and probe energies were measured and also the effect of detector gain, the gate delay and the gate width were studied. In

  12. Signal enhancement by a multi-layered substrate for mutagen detection using an SOS response-induced green fluorescent protein in genetically modified Escherichia coli.

    Science.gov (United States)

    Etoh, Hiroki; Yasuda, Mitsuru; Akimoto, Takuo

    2011-01-01

    In this paper, we describe a method to enhance the fluorescence signal of mutagen detection using SOS response-induced green fluorescence protein (GFP) in genetically modified Escherichia coli using a multi-layered substrate. To generate E. coli that express SOS response-induced GFP, we constructed a plasmid carrying the RecA promoter located upstream of the GFP gene and used it to transform E. coli BL21. The transformed strain was incubated with mitomycin C (MMC), a typical mutagen, and then immobilized on a multi-layered substrate with Ag and a thin Al(2)O(3) layer on a glass slide. Since the multi-layered substrate technique is an optical technique with potential to enhance the fluorescence of fluorophore placed on top of the substrate, the multi-layered substrate was expected to improve the fluorescence signal of mutagen detection. We obtained an average 14-fold fluorescence enhancement of MMC-induced GFP in the concentration range 1 to 1000 ng/ml. In addition, the lower detection limit of MMC was improved using this technique, and was estimated to be 1 ng/ml because of an enlargement of the difference between the blank and the signal of 1 ng/ml of MMC.

  13. Tracking of Mesenchymal Stem Cells with Fluorescence Endomicroscopy Imaging in Radiotherapy-Induced Lung Injury

    Science.gov (United States)

    Perez, Jessica R.; Ybarra, Norma; Chagnon, Frederic; Serban, Monica; Lee, Sangkyu; Seuntjens, Jan; Lesur, Olivier; El Naqa, Issam

    2017-01-01

    Mesenchymal stem cells (MSCs) have potential for reducing inflammation and promoting organ repair. However, limitations in available techniques to track them and assess this potential for lung repair have hindered their applicability. In this work, we proposed, implemented and evaluated the use of fluorescence endomicroscopy as a novel imaging tool to track MSCs in vivo. MSCs were fluorescently labeled and injected into a rat model of radiation-induced lung injury via endotracheal (ET) or intravascular (IV) administration. Our results show that MSCs were visible in the lungs with fluorescence endomicroscopy. Moreover, we developed an automatic cell counting algorithm to quantify the number of detected cells in each condition. We observed a significantly higher number of detected cells in ET injection compared to IV and a slight increase in the mean number of detected cells in irradiated lungs compared to control, although the latter did not reach statistical significance. Fluorescence endomicroscopy imaging is a powerful new minimally invasive and translatable tool that can be used to track and quantify MSCs in the lungs and help assess their potential in organ repair.

  14. In vivo Diagnosis of Cervical Intraepithelial Neoplasia Using 337-nm- Excited Laser-Induced Fluorescence

    Science.gov (United States)

    Ramanujam, N.; Mitchell, M. F.; Mahadevan, A.; Warren, S.; Thomsen, S.; Silva, E.; Richards-Kortum, R.

    1994-10-01

    Laser-induced fluorescence at 337-nm excitation was used in vivo to differentiate neoplastic [cervical intraepithelial neoplasia (CIN)], nonneoplastic abnormal (inflammation and human papilloma viral infection), and normal cervical tissues. A colposcope (low-magnification microscope used to view the cervix with reflected light) was used to identify 66 normal and 49 abnormal (5 inflammation, 21 human papilloma virus infection, and 23 CIN) sites on the cervix in 28 patients. These sites were then interrogated spectroscopically. A two-stage algorithm was developed to diagnose CIN. The first stage differentiated histologically abnormal tissues from colposcopically normal tissues with a sensitivity, specificity, and positive predictive value of 92%, 90%, and 88%, respectively. The second stage differentiated preneoplastic and neoplastic tissues from nonneoplastic abnormal tissues with a sensitivity, specificity, and positive predictive value of 87%, 73%, and 74%, respectively. Spectroscopic differences were consistent with a decrease in the absolute contribution of collagen fluorescence, an increase in the absolute contribution of oxyhemoglobin attenuation, and an increase in the relative contribution of reduced nicotinamide dinucleotide phosphate [NAD(P)H] fluorescence as tissue progresses from normal to abnormal in the same patient. These results suggest that in vivo fluorescence spectroscopy of the cervix can be used to diagnose CIN at colposcopy.

  15. Tunable KrF laser-induced fluorescence of C2 in a sooting flame

    Science.gov (United States)

    Brockhinke, A.; Hartlieb, A. T.; Kohse-Höinghaus, K.; Crosley, D. R.

    1998-11-01

    We have observed laser-induced fluorescence of two different band systems of C2 in a flame, excited by a tunable KrF laser near 248 nm. The first comprises several P and R lines of the (1,0) band of the e 3@g-a 3@u Fox-Herzberg system, with fluorescence bands extending past 350 nm. The second is the band head region of the (7,1) band of the D 1Du+LB' 1Dg+ system, with fluorescence at 232 nm from D to the X 1Dg+ ground state. Neither band has been previously observed in any environment. The flame in these experiments is highly sooting, and the C2 seen here is likely produced by laser vaporization of the soot with subsequent laser photolysis of a C2 precursor. In a rich flame, this fluorescence could cause interferences in other studies such as KrF laser Raman scattering. Moreover, signal level calculations suggest native C2 near 10 ppm could be readily observed using the Fox-Herzberg excitation. Raman measurements of major species (X̾.01) in the same flame, using the KrF laser, are in good agreement with a model prediction.

  16. Near-field measurements of vegetation by laser-induced fluorescence imaging

    Science.gov (United States)

    Sowinska, Malgorzata; Cunin, Bernard; Deruyver, Aline; Heisel, Francine; Miehe, Joseph-Albert; Langsdorf, Gabriele; Lichtenthaler, Hartmut K.

    1999-12-01

    In this paper, a validation of a new UV-A laser-induced fluorescence imaging system implemented in an all-road car for near-field remote sensing of vegetation will be presented. It has been developed as a part of a European Community Program INTERREG II and is consisting of three main parts: excitation, detection and control units. The excitation source is a frequency tripled Nd:YAG laser and the laser spot size is adjusted via a variable beam expander. Fluorescence images are recorded at four characteristic fluorescence bands: 440, 520, 690 and 740 nm with a gated intensified digital CCD camera. The laser head and camera are situated on a directed in site and azimuth platform which can be high up to 6 meters. The platform positioning, localization and distance detection, spot size determination and adjustment, focus, sharpness, selection of the filter, laser and camera synchronization, gain of the intensifier, real time visualization of images, acquisition time are controlled by a newly developed software which allows also image storage, analysis and treatment. Examples of remote sensing fluorescence images from several plant species recorded at a distance of 10 - 30 m will be given and discussed further in this paper.

  17. Tracking of Mesenchymal Stem Cells with Fluorescence Endomicroscopy Imaging in Radiotherapy-Induced Lung Injury

    Science.gov (United States)

    Perez, Jessica R.; Ybarra, Norma; Chagnon, Frederic; Serban, Monica; Lee, Sangkyu; Seuntjens, Jan; Lesur, Olivier; El Naqa, Issam

    2017-01-01

    Mesenchymal stem cells (MSCs) have potential for reducing inflammation and promoting organ repair. However, limitations in available techniques to track them and assess this potential for lung repair have hindered their applicability. In this work, we proposed, implemented and evaluated the use of fluorescence endomicroscopy as a novel imaging tool to track MSCs in vivo. MSCs were fluorescently labeled and injected into a rat model of radiation-induced lung injury via endotracheal (ET) or intravascular (IV) administration. Our results show that MSCs were visible in the lungs with fluorescence endomicroscopy. Moreover, we developed an automatic cell counting algorithm to quantify the number of detected cells in each condition. We observed a significantly higher number of detected cells in ET injection compared to IV and a slight increase in the mean number of detected cells in irradiated lungs compared to control, although the latter did not reach statistical significance. Fluorescence endomicroscopy imaging is a powerful new minimally invasive and translatable tool that can be used to track and quantify MSCs in the lungs and help assess their potential in organ repair. PMID:28102237

  18. Femtosecond study of light-induced fluorescence increase of the dark chromoprotein asFP595

    Energy Technology Data Exchange (ETDEWEB)

    Schuettrigkeit, Tanja A. [Department Chemie, Technische Universitaet Muenchen, 85747 Garching (Germany); Feilitzsch, Till von [Department Chemie, Technische Universitaet Muenchen, 85747 Garching (Germany); Kompa, Christian K. [Department Chemie, Technische Universitaet Muenchen, 85747 Garching (Germany); Lukyanov, Konstantin A. [Shemiakin and Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Moscow (Russian Federation); Savitsky, Alexander P. [A.N. Bach Institute of Biochemistry, Russian Academy of Sciences, Moscow (Russian Federation); Voityuk, Alexander A. [Institucio Catalana de Recerca i Estudis Avancats (ICREA), Institute of Computational Chemistry, Universitat de Girona (Spain); Michel-Beyerle, Maria E. [Department Chemie, Technische Universitaet Muenchen, 85747 Garching (Germany)], E-mail: michel-beyerle@ch.tum.de

    2006-04-21

    Femtosecond time-resolved spectroscopy is applied to study the mechanism of the light-induced increase of fluorescence quantum yield of the initially non-fluorescent (dark) chromoprotein asFP595. Spectroscopic and kinetic characteristics of this unique fluorescence 'kindling' phenomenon are: (i) the small Stokes shift of the dark chromophore consistent with either the zwitterion or the anion; (ii) the singlet excited state of the dark chromophore decaying predominantly with a time constant of {approx}320 fs corresponding to a fluorescence quantum yield {phi} {sub Fl} {<=} 10{sup -4}. Since ground state recovery occurs on the same time scale, this radiationless channel is assigned to internal conversion; (iii) the formation of the fluorescent species depending on the sequential absorption of two photons with a delay significantly exceeding the excitation pulse duration of 150 fs; (iv) the fluorescent species showing a red-shift of {approx}20 nm in absorption and emission, and an excited state lifetime of 2.2 ns. The ultrafast internal conversion of the excited dark state is attributed to the proximity of the S{sub 0} and S{sub 1} potential energy surfaces favored by the non-planarity of the chromophore as revealed in recent X-ray structures. Competing with internal conversion two different transformations of the chromophore structure are suggested which may be identified in a future X-ray structural analysis of the the photoconverted fluorescent state. The predominant kindling mechanism may be either (i) trans-cis isomerization or (ii) proton transfer between an excited zwitterion and the protein cleft. For mechanism (ii) the large dipole moment change of about 11 D upon S{sub 0}-S{sub 1} excitation of the chromophore would be crucial in order to initiate protein relaxation and deprotonation of a zwitterion. Both mechanisms are assumed to lead to a metastable planar structure responsible for the long-lived fluorescence of the chromophore &apos

  19. High-speed confocal fluorescence lifetime imaging microscopy by analog mean-delay method

    Science.gov (United States)

    Won, Youngjae; Kim, Donguk; Yang, Wenzhong; Kim, Dug Y.

    2010-02-01

    We have demonstrated the high-speed confocal fluorescence lifetime imaging microscopy (FLIM) by analog mean-delay (AMD) method. The AMD method is a new signal processing technique for calculation of fluorescence lifetime and it is very suitable for the high-speed confocal FLIM with good accuracy and photon economy. We achieved the acquisition speed of 7.7 frames per second for confocal FLIM imaging. Here, the highest photon detection rate for one pixel was larger than 125 MHz and averaged photon detection rate was more than 62.5 MHz. Based on our system, we successfully obtained a sequence of confocal fluorescence lifetime images of RBL-2H3 cell labeled with Fluo-3/AM and excited by 4αPDD (TRPV channel agonist) within one second.

  20. Applicability of UV laser-induced solid-state fluorescence spectroscopy for characterization of solid dosage forms.

    Science.gov (United States)

    Woltmann, Eva; Meyer, Hans; Weigel, Diana; Pritzke, Heinz; Posch, Tjorben N; Kler, Pablo A; Schürmann, Klaus; Roscher, Jörg; Huhn, Carolin

    2014-10-01

    High production output of solid pharmaceutical formulations requires fast methods to ensure their quality. Likewise, fast analytical procedures are required in forensic sciences, for example at customs, to substantiate an initial suspicion. We here present the design and the optimization of an instrumental setup for rapid and non-invasive characterization of tablets by laser-induced fluorescence spectroscopy (with a UV-laser (λ ex = 266 nm) as excitation source) in reflection geometry. The setup was first validated with regard to repeatability, bleaching phenomena, and sensitivity. The effect on the spectra by the physical and chemical properties of the samples, e.g. their hardness, homogeneity, chemical composition, and granule grain size of the uncompressed material, using a series of tablets, manufactured in accordance with design of experiments, was investigated. Investigation of tablets with regard to homogeneity, especially, is extremely important in pharmaceutical production processes. We demonstrate that multiplicative scatter correction is an appropriate tool for data preprocessing of fluorescence spectra. Tablets with different physical and chemical characteristics can be discriminated well from their fluorescence spectra by subjecting the results to principal component analysis.

  1. Interactions of a lytic peptide with supported lipid bilayers investigated by time-resolved evanescent wave-induced fluorescence spectroscopy

    Science.gov (United States)

    Rapson, Andrew C.; Gee, Michelle L.; Clayton, Andrew H. A.; Smith, Trevor A.

    2016-12-01

    We report investigations, using time-resolved and polarised evanescent wave-induced fluorescence methods, into the location, orientation and mobility of a fluorescently labelled form of the antimicrobial peptide, melittin, when it interacts with vesicles and supported lipid bilayers (SLBs). This melittin analogue, termed MK14-A430, was found to penetrate the lipid headgroup structure in pure, ordered-phase DPPC membranes but was located near the headgroup-water region when cholesterol was included. MK14-A430 formed lytic pores in SLBs, and an increase in pore formation with incubation time was observed through an increase in polarity and mobility of the probe. When associated with the Cholesterol-containing SLB, the probe displayed polarity and mobility that indicated a population distributed near the lipid headgroup-water interface with MK14-A430 arranged predominantly in a surface-aligned state. This study indicates that the lytic activity of MK14-A430 occurred through a pore-forming mechanism. The lipid headgroup environment experienced by the fluorescent label, where MK14-A430 displayed pore information, indicated that pore formation was best described by the toroidal pore model.

  2. Miniaturized fluorescent RNA dot blot method for rapid quantitation of gene expression

    Directory of Open Access Journals (Sweden)

    Yadetie Fekadu

    2004-06-01

    Full Text Available Abstract Background RNA dot blot hybridization is a commonly used technique for gene expression assays. However, membrane based RNA dot/slot blot hybridization is time consuming, requires large amounts of RNA, and is less suited for parallel assays of more than one gene at a time. Here, we describe a glass-slide based miniaturized RNA dot blot (RNA array procedure for rapid and parallel gene expression analysis using fluorescently labeled probes. Results RNA arrays were prepared by simple manual spotting of RNA onto amino-silane coated microarray glass slides, and used for two-color fluorescent hybridization with specific probes labeled with Cy3 and 18S ribosomal RNA house-keeping gene probe labeled with Cy5 fluorescent dyes. After hybridization, arrays were scanned on a fluorescent microarray scanner and images analyzed using microarray image analysis software. We demonstrate that this method gives comparable results to Northern blot analysis, and enables high throughput quantification of transcripts from nanogram quantities of total RNA in hundreds of samples. Conclusion RNA array on glass slide and detection by fluorescently labeled probes can be used for rapid and parallel gene expression analysis. The method is particularly well suited for gene expression assays that involve quantitation of many transcripts in large numbers of samples.

  3. The drought impact on satellite solar-induced chlorophyll fluorescence in China during 2007-2015

    Science.gov (United States)

    Li, Ruitao

    2016-04-01

    Drought is one of the most damaging and complicated natural hazards in the world. China is one of the countries which are most severely affected by drought. And there is a severe drought event in China every 2-3 years. From the beginning of the 1980s, some vegetation indices have been used to monitor vegetation under water stress. With the development of remote sensing technology, satellite solar-induced chlorophyll fluorescence (SIF) has emerged as a new method to monitor vegetation in recent years. Some studies have shown that compared with vegetation indices, SIF is more sensitive for vegetation functioning. However, the related studies using the satellite SIF is relatively limited in China. The objective of this study is to investigate the impact of drought on SIF by analyzing the relationships of SIF and crucial land surface parameter under the drought condition and to assess the adaption of satellite SIF in China. The SIF data are from the Global Ozone Monitoring Experiment 2 (GOME-2). Firstly, the widely used Palmer Drought Severity Index (PDSI) was used for drought events identification from 2007 to 2015 in China. On the basis of the identification results, we chose a number of areas of interest according to different land cover types and drought intensity. Then, we analyzed the relationships of SIF and land surface variables, i.e. normalized difference vegetation index (NDVI), the fraction of absorbed photosynthetically active radiation (fPAR), root-zone soil moisture (SMC) and surface skin temperatures (Tskin). The results show that the spatial patterns of negative SIF anomalies are closely relevant to the drought intensity. The decrease of SIF is aggravated in the phase of drought occurs. Moreover we find that the GOME-2 SIF is sensitive to fPAR and fluorescence yield. And the SIF is strongly correlated with SMC, Tskin and NDVI. But the SIF decreases more rapidly during the early time of drought events than NDVI. In other words, the SIF can well capture

  4. An Automated Comparative Observation System for Sun-Induced Chlorophyll Fluorescence of Vegetation Canopies

    Directory of Open Access Journals (Sweden)

    Xijia Zhou

    2016-05-01

    Full Text Available Detecting sun-induced chlorophyll fluorescence (SIF offers a new approach for remote sensing photosynthesis. However, to analyse the response characteristics of SIF under different stress states, a long-term time-series comparative observation of vegetation under different stress states must be carried out at the canopy scale, such that the similarities and differences in SIF change law can be summarized under different time scales. A continuous comparative observation system for vegetation canopy SIF is designed in this study. The system, which is based on a high-resolution spectrometer and an optical multiplexer, can achieve comparative observation of multiple targets. To simultaneously measure the commonly used vegetation index and SIF in the O2-A and O2-B atmospheric absorption bands, the following parameters are used: a spectral range of 475.9 to 862.2 nm, a spectral resolution of approximately 0.9 nm, a spectral sampling interval of approximately 0.4 nm, and the signal-to-noise ratio (SNR can be as high as 1000:1. To obtain data for both the upward radiance of the vegetation canopy and downward irradiance data with a high SNR in relatively short time intervals, the single-step integration time optimization algorithm is proposed. To optimize the extraction accuracy of SIF, the FluorMOD model is used to simulate sets of data according to the spectral resolution, spectral sampling interval and SNR of the spectrometer in this continuous observation system. These data sets are used to determine the best parameters of Fraunhofer Line Depth (FLD, Three FLD (3FLD and the spectral fitting method (SFM, and 3FLD and SFM are confirmed to be suitable for extracting SIF from the spectral measurements. This system has been used to observe the SIF values in O2-A and O2-B absorption bands and some commonly used vegetation index from sweet potato and bare land, the result of which shows: (1 the daily variation trend of SIF value of sweet potato leaves is

  5. An Automated Comparative Observation System for Sun-Induced Chlorophyll Fluorescence of Vegetation Canopies.

    Science.gov (United States)

    Zhou, Xijia; Liu, Zhigang; Xu, Shan; Zhang, Weiwei; Wu, Jun

    2016-05-27

    Detecting sun-induced chlorophyll fluorescence (SIF) offers a new approach for remote sensing photosynthesis. However, to analyse the response characteristics of SIF under different stress states, a long-term time-series comparative observation of vegetation under different stress states must be carried out at the canopy scale, such that the similarities and differences in SIF change law can be summarized under different time scales. A continuous comparative observation system for vegetation canopy SIF is designed in this study. The system, which is based on a high-resolution spectrometer and an optical multiplexer, can achieve comparative observation of multiple targets. To simultaneously measure the commonly used vegetation index and SIF in the O₂-A and O₂-B atmospheric absorption bands, the following parameters are used: a spectral range of 475.9 to 862.2 nm, a spectral resolution of approximately 0.9 nm, a spectral sampling interval of approximately 0.4 nm, and the signal-to-noise ratio (SNR) can be as high as 1000:1. To obtain data for both the upward radiance of the vegetation canopy and downward irradiance data with a high SNR in relatively short time intervals, the single-step integration time optimization algorithm is proposed. To optimize the extraction accuracy of SIF, the FluorMOD model is used to simulate sets of data according to the spectral resolution, spectral sampling interval and SNR of the spectrometer in this continuous observation system. These data sets are used to determine the best parameters of Fraunhofer Line Depth (FLD), Three FLD (3FLD) and the spectral fitting method (SFM), and 3FLD and SFM are confirmed to be suitable for extracting SIF from the spectral measurements. This system has been used to observe the SIF values in O₂-A and O₂-B absorption bands and some commonly used vegetation index from sweet potato and bare land, the result of which shows: (1) the daily variation trend of SIF value of sweet potato leaves is

  6. Stable and Size-Tunable Aggregation-Induced Emission Nanoparticles Encapsulated with Nanographene Oxide and Applications in Three-Photon Fluorescence Bioimaging.

    Science.gov (United States)

    Zhu, Zhenfeng; Qian, Jun; Zhao, Xinyuan; Qin, Wei; Hu, Rongrong; Zhang, Hequn; Li, Dongyu; Xu, Zhengping; Tang, Ben Zhong; He, Sailing

    2016-01-26

    Organic fluorescent dyes with high quantum yield are widely applied in bioimaging and biosensing. However, most of them suffer from a severe effect called aggregation-caused quenching (ACQ), which means that their fluorescence is quenched at high molecular concentrations or in the aggregation state. Aggregation-induced emission (AIE) is a diametrically opposite phenomenon to ACQ, and luminogens with this feature can effectively solve this problem. Graphene oxide has been utilized as a quencher for many fluorescent dyes, based on which biosensing can be achieved. However, using graphene oxide as a surface modification agent of fluorescent nanoparticles is seldom reported. In this article, we used nanographene oxide (NGO) to encapsulate fluorescent nanoparticles, which consisted of a type of AIE dye named TPE-TPA-FN (TTF). NGO significantly improved the stability of nanoparticles in aqueous dispersion. In addition, this method could control the size of nanoparticles' flexibly as well as increase their emission efficiency. We then used the NGO-modified TTF nanoparticles to achieve three-photon fluorescence bioimaging. The architecture of ear blood vessels in mice and the distribution of nanoparticles in zebrafish could be observed clearly. Furthermore, we extended this method to other AIE luminogens and showed it was widely feasible.

  7. Measurement of radiative lifetime in atomic samarium using simultaneous detection of laser-induced fluorescence and photoionization signals

    Indian Academy of Sciences (India)

    A C Sahoo; M L Shah; P K Mandal; A K Pulhani; G P Gupta; Vas Dev; B M Suri

    2014-02-01

    In this paper, we report the investigations of lifetime measurement of odd-parity energy level 19009.52 cm-1 of Sm I using simultaneous detection of laser-induced fluorescence and laserinduced photoionization signals employing pump–probe technique. To the best of our knowledge, this is for the first time that the results obtained using laser-induced fluorescence and photoionization techniques have been compared with each other. The obtained results match well with those reported in the literature.

  8. Improving Resolution of Confocal Laser Induced Fluorescence in Argon Helicon Plasma

    Science.gov (United States)

    Soderholm, Mark; Vandervort, Robert; Scime, Earl; McKee, John; McCarren, Dustin

    2014-10-01

    Laser Induced Fluorescence (LIF) provides measurements of flow speed, temperature and when absolutely calibrated, density of ions or neutrals in a plasma. Traditionally, laser induced fluorescence requires two ports on a plasma device. One port is used for laser injection and the other is used for fluorescence emission collection. Traditional LIF is tedious and time consuming to align. These difficulties motivate the development of an optical configuration that requires a single port and remains fully aligned at all times; confocal LIF. Our confocal optical design employs a single two inch diameter lens to both inject the laser light and collect the stimulated emission from an argon plasma. A dichroic mirror is used to separate the injected laser light from the collected emission. The measurement location is scanned radially by manually adjusting the final focusing lens position. In the initial version of the confocal optical system, measurements were poorly resolved radially because they were integrated over a fairly large path length (~4 cm) centered at the focal point. Here we present collected data from a modified configuration that significantly improves the special resolution of confocal measurements. The confocal measurements are compared to traditional, two-port, LIF measurements over the same radial range.

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

    Science.gov (United States)

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

    2015-09-01

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

  10. Polyamines induce aggregation of LHC II and quenching of fluorescence in vitro.

    Science.gov (United States)

    Tsiavos, Theodoros; Ioannidis, Nikolaos E; Kotzabasis, Kiriakos

    2012-05-01

    Dissipation of excess excitation energy within the light-harvesting complex of Photosystem II (LHC II) is a main process in plants, which is measured as the non-photochemical quenching of chlorophyll fluorescence or qE. We showed in previous works that polyamines stimulate qE in higher plants in vivo and in eukaryotic algae in vitro. In the present contribution we have tested whether polyamines can stimulate quenching in trimeric LHC II and monomeric light-harvesting complex b proteins from higher plants. The tetramine spermine was the most potent quencher and induced aggregation of LHC II trimers, due to its highly cationic character. Two transients are evident at 100 μM and 350 μM for the fluorescence and absorbance signals of LHC II respectively. On the basis of observations within this work, some links between polyamines and the activation of qE in vivo is discussed.

  11. Compact low-cost detector for in vivo assessment of microphytobenthos using laser induced fluorescence

    Science.gov (United States)

    Utkin, A. B.; Vieira, S.; Marques da Silva, J.; Lavrov, A.; Leite, E.; Cartaxana, P.

    2013-03-01

    The development of a compact low-cost detector for non-destructive assessment of microphytobenthos using laser induced fluorescence was described. The detector was built from a specially modified commercial miniature fiber optic spectrometer (Ocean Optics USB4000). Its usefulness is experimentally verified by the study of diatom-dominated biofilms inhabiting the upper layers of intertidal sediments of the Tagus Estuary, Portugal. It is demonstrated that, operating with a laser emitter producing 30 mJ pulses at the wavelength of 532 nm, the detector is capable to record fluorescence signals with sufficient intensity for the quantitative biomass characterization of the motile epipelic microphytobenthic communities and to monitor their migratory activity. This paves the way for building an entire emitter-detector LIF system for microphytobenthos monitoring, which will enable microalgae communities occupying hardly accessible intertidal flats to be monitored in vivo at affordable cost.

  12. A LabVIEW-Based Virtual Instrument System for Laser-Induced Fluorescence Spectroscopy.

    Science.gov (United States)

    Wu, Qijun; Wang, Lufei; Zu, Lily

    2011-01-01

    We report the design and operation of a Virtual Instrument (VI) system based on LabVIEW 2009 for laser-induced fluorescence experiments. This system achieves synchronous control of equipment and acquisition of real-time fluorescence data communicating with a single computer via GPIB, USB, RS232, and parallel ports. The reported VI system can also accomplish data display, saving, and analysis, and printing the results. The VI system performs sequences of operations automatically, and this system has been successfully applied to obtain the excitation and dispersion spectra of α-methylnaphthalene. The reported VI system opens up new possibilities for researchers and increases the efficiency and precision of experiments. The design and operation of the VI system are described in detail in this paper, and the advantages that this system can provide are highlighted.

  13. Application of quantitative light-induced fluorescence to determine the depth of demineralization of dental fluorosis in enamel microabrasion: a case report

    OpenAIRE

    PARK, Tae-Young; Choi, Han-Sol; Ku, Hee-Won; Kim, Hyun-Su; Lee, Yoo-Jin; Min, Jeong-Bum

    2016-01-01

    Enamel microabrasion has become accepted as a conservative, nonrestorative method of removing intrinsic and superficial dysmineralization defects from dental fluorosis, restoring esthetics with minimal loss of enamel. However, it can be difficult to determine if restoration is necessary in dental fluorosis, because the lesion depth is often not easily recognized. This case report presents a method for analysis of enamel hypoplasia that uses quantitative light-induced fluorescence (QLF) follow...

  14. Base pairing enhances fluorescence and favors cyclobutane dimer formation induced upon absorption of UVA radiation by DNA.

    Science.gov (United States)

    Banyasz, Akos; Vayá, Ignacio; Changenet-Barret, Pascale; Gustavsson, Thomas; Douki, Thierry; Markovitsi, Dimitra

    2011-04-13

    The photochemical properties of the DNA duplex (dA)(20)·(dT)(20) are compared with those of the parent single strands. It is shown that base pairing increases the probability of absorbing UVA photons, probably due to the formation of charge-transfer states. UVA excitation induces fluorescence peaking at ∼420 nm and decaying on the nanosecond time scale. The fluorescence quantum yield, the fluorescence lifetime, and the quantum yield for cyclobutane dimer formation increase upon base pairing. Such behavior contrasts with that of the UVC-induced processes.

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

    Energy Technology Data Exchange (ETDEWEB)

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

    2014-01-15

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

  16. Method for detecting binding events using micro-X-ray fluorescence spectrometry

    Science.gov (United States)

    Warner, Benjamin P.; Havrilla, George J.; Mann, Grace

    2010-12-28

    Method for detecting binding events using micro-X-ray fluorescence spectrometry. Receptors are exposed to at least one potential binder and arrayed on a substrate support. Each member of the array is exposed to X-ray radiation. The magnitude of a detectable X-ray fluorescence signal for at least one element can be used to determine whether a binding event between a binder and a receptor has occurred, and can provide information related to the extent of binding between the binder and receptor.

  17. Fluorescence quenching method for the determination of catechol with gold nanoparticles and tyrosinase hybrid system

    Institute of Scientific and Technical Information of China (English)

    Martin; M.F.Choi

    2010-01-01

    The determination method of catechol by fluorescence quenching was developed.The assay was based on the combination of the unique property of gold nanoparticles with tyrosinase enzymatic reaction.In the presence of tyrosinase,the fluorescence of gold nanoparticles was quenched by catechol which can be employed to detect catechol.Under the optimal conditions,a linear range 5.0×10~(-7)-1.0×10~(-3) mol L~(-1) and a detection limit 1.0×10~(-7) mol L~(-1) of catechol were obtained.o-Quinone intermediate produ...

  18. Characterisation of an inlet pre-injector laser induced fluorescence instrument for the measurement of ambient hydroxyl radicals

    Directory of Open Access Journals (Sweden)

    A. Novelli

    2014-01-01

    Full Text Available Ambient measurements of hydroxyl radicals (OH are challenging due to a high reactivity and consequently low concentration. The importance of OH as an atmospheric oxidant has resulted in a sustained effort leading to the development of a number of analytical techniques. Recent work has indicated that the laser-induced fluorescence of the OH molecules method based on the fluorescence assay by gas expansion technique (LIF-FAGE for the measurement of atmospheric OH in some environments may be influenced by artificial OH generated within the instrument, and a chemical method to remove this interference was implemented in a LIF-FAGE system by Mao et al. (2012. We have applied this method to our LIF-FAGE HORUS (HydrOxyl Radical Measurement Unit based on fluorescence Spectroscopy system, and developed and deployed an inlet pre-injector (IPI to determine the chemical zero level in the instrument via scavenging the ambient OH radical. We describe and characterise this technique in addition to its application at field sites in forested locations in Finland, Spain, and Germany. Ambient measurements show that OH generated within the HORUS instrument is a non-negligible fraction of the total OH signal, which can comprise 30% to 80% during the day and 60% to 100% during the night. The contribution of the background OH varied greatly between measurement sites and was likely related to the type and concentration of volatile organic compounds (VOCs present at each particular location. Two inter-comparisons in contrasting environments between the HORUS instrument and two different chemical ionisation mass spectrometers (CIMS are described to demonstrate the efficacy of the inlet-pre-injector and the necessity of the chemical zeroing method in such environments.

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

    DEFF Research Database (Denmark)

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

    2013-01-01

    This mini-review reports on applications of particular multiphoton excitation microscopy-based methodologies employed in our laboratory to study skin. These approaches allow in-depth optical sectioning of the tissue, providing spatially resolved information on specific fluorescence probes......' parameters. Specifically, by applying these methods, spatially resolved maps of water dipolar relaxation (generalized polarization function using the 6-lauroyl-2-(N,N-dimethylamino)naphthale probe), activity of protons (fluorescence lifetime imaging using a proton sensitive fluorescence probe--2,7-bis-(2...... excised skin, including applications of fluctuation correlation spectroscopy on transdermal penetration of liposomes are presented and discussed. The data from the different studies reported reveal the intrinsic heterogeneity of skin and also prove these strategies to be powerful noninvasive tools...

  20. Determination of L-phenylalanine by cucurbit[7]uril sensitized fluorescence quenching method

    Institute of Scientific and Technical Information of China (English)

    Chang Feng Li; Li Ming Du; Hao Wu; Ying Xia Chang

    2011-01-01

    The determination method of L-phenylalanine (LPA) by fluorescence quenching was developed. The assay was based on the combination of the cucurbit[7]uril (CB[7]) with palmatine hydrochloride (PAL) reaction. In the presence of CB[7], the fluorescence of PAL was quenched by LPA which can be employed to detect LPA. Under the optimal conditions, a linear range 3.63 × 10-8-9.68 × 10-6 mol/L and a detection limit 1.27 × 10-8 mol/L of LPA were obtained. The relative standard deviation (R.S.D) was 1.8% obtained from a series of11 standards each containing 6.05 × 10-6 mol/L oLPA. This paper also discusses the mechanism of fluorescence indicator probe.

  1. Concentration Measurements in a Cold Flow Model Annular Combustor Using Laser Induced Fluorescence

    Science.gov (United States)

    Morgan, Douglas C.

    1996-01-01

    A nonintrusive concentration measurement method is developed for determining the concentration distribution in a complex flow field. The measurement method consists of marking a liquid flow with a water soluble fluorescent dye. The dye is excited by a two dimensional sheet of laser light. The fluorescent intensity is shown to be proportional to the relative concentration level. The fluorescent field is recorded on a video cassette recorder through a video camera. The recorded images are analyzed with image processing hardware and software to obtain intensity levels. Mean and root mean square (rms) values are calculated from these intensity levels. The method is tested on a single round turbulent jet because previous concentration measurements have been made on this configuration by other investigators. The previous results were used to comparison to qualify the current method. These comparisons showed that this method provides satisfactory results. 'Me concentration measurement system was used to measure the concentrations in the complex flow field of a model gas turbine annular combustor. The model annular combustor consists of opposing primary jets and an annular jet which discharges perpendicular to the primary jets. The mixing between the different jet flows can be visualized from the calculated mean and rms profiles. Concentration field visualization images obtained from the processing provide further qualitative information about the flow field.

  2. Fluorescence measurements of serotonin-induced V-ATPase-dependent pH changes at the luminal surface in salivary glands of the blowfly Calliphora vicina.

    Science.gov (United States)

    Rein, Julia; Zimmermann, Bernhard; Hille, Carsten; Lang, Ingo; Walz, Bernd; Baumann, Otto

    2006-05-01

    Secretion in blowfly salivary glands is induced by the neurohormone serotonin and powered by a vacuolar-type H(+)-ATPase (V-ATPase) located in the apical membrane of the secretory cells. We have established a microfluorometric method for analysing pH changes at the luminal surface of the secretory epithelial cells by using the fluorescent dye 5-N-hexadecanoyl-aminofluorescein (HAF). After injection of HAF into the lumen of the tubular salivary gland, the fatty acyl chain of the dye molecule partitions into the outer leaflet of the plasma membrane and its pH-sensitive fluorescent moiety is exposed at the cell surface. Confocal imaging has confirmed that HAF distributes over the entire apical membrane of the secretory cells and remains restricted to this membrane domain. Ratiometric analysis of HAF fluorescence demonstrates that serotonin leads to a reversible dose-dependent acidification at the luminal surface. Inhibition by concanamycin A confirms that the serotonin-induced acidification at the luminal surface is due to H(+) transport across the apical membrane via V-ATPase. Measurements with pH-sensitive microelectrodes corroborate a serotonin-induced luminal acidification and demonstrate that luminal pH decreases by about 0.4 pH units at saturating serotonin concentrations. We conclude that ratiometric measurements of HAF fluorescence provide an elegant method for monitoring V-ATPase-dependent H(+) transport in the blowfly salivary gland in vivo and for analysing the spatiotemporal pattern of pH changes at the luminal surface.

  3. PhotoSpec - Ground-based Remote Sensing of Solar-Induced Chlorophyll Fluorescence: First Results

    Science.gov (United States)

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

    2016-12-01

    Solar-Induced Chlorophyll Fluorescence (SIF) emitted from vegetation can be used as a proxy for photosynthetic activity and is 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 from environmental conditions, water stress, or radiation. We have developed a novel ground-based spectrometer system for measuring SIF from natural ecosystems. 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, and also includes a commercial photosynthetic active radiation (PAR) sensor. The spectrometers cover a SIF retrieval wavelength range at high spectral resolution (670 - 780 nm, 0.1 nm FWHM), and also provide moderate resolution spectra (400 - 800 nm, 1.5 nm FWHM) to retrieve vegetation indices and the photochemical reflectance index (PRI). We report on results of the first continuous field measurements of this novel system at Stunt Ranch Santa Monica Mountains UC Reserve, where the PhotoSpec instrument was monitoring SIF of four native Californian shrubland species with different adaptations to seasonal summer drought. We report on the correlation with CO2 fluxes over both the growing season and the hot summer period in 2016. We also show detailed measurements of the diurnal cycle of the SIF signal of single broad leaves, as well as dark-light transitions, under controlled experimental conditions. In addition to demonstrating the instrumental set-up, retrieval algorithm, and instrument performance, our results illustrate that SIF measurements at the leaf to ecosystem scale are needed to understand and interpret the SIF signals retrieved at larger scales.

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

    Energy Technology Data Exchange (ETDEWEB)

    Neij, H.

    1998-11-01

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

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

    Energy Technology Data Exchange (ETDEWEB)

    Neij, H.

    1998-11-01

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

  6. A novel fluorescence derivatization method combined with HPLC for determining the activities of endogenous caspase.

    Science.gov (United States)

    Liu, Jiachi; Lu, Ye; Liang, Jianying

    2012-11-07

    A novel fluorescence derivatization method combined with HPLC was developed to detect the activity of caspase-3 and -8 in two cell lines (Hela cells and A549 cells) which were activated by low temperature-assisted ultraviolet irradiation (LT-UV), mitomycin C (MMC) and camptothecin during the apoptosis, respectively. Two peptide substrates for either caspase-3 or -8 were designed, of which peptide fragments were obtained by enzymatic modification, followed by fluorescence derivatization. A single fluorescent product was formed when a peptide was heated at 120 °C for 10 min in a neutral aqueous medium (pH 7.0) containing catechol, sodium periodate and sodium borate. Commercial kits for detecting the activity of caspase-3 and -8 were used as a control. The relative activity of the caspases detected by fluorescence derivatization was similar to that obtained by commercial kits, which indicated that the novel method is reliable. The activity assays of recombinant human caspases showed that the novel method provided higher selectivity than that of commercial kits, which proved it to be more accurate for determining the activity of caspases in apoptosis.

  7. Quantitative method to assess caries via fluorescence imaging from the perspective of autofluorescence spectral analysis

    Science.gov (United States)

    Chen, Q. G.; Zhu, H. H.; Xu, Y.; Lin, B.; Chen, H.

    2015-08-01

    A quantitative method to discriminate caries lesions for a fluorescence imaging system is proposed in this paper. The autofluorescence spectral investigation of 39 teeth samples classified by the International Caries Detection and Assessment System levels was performed at 405 nm excitation. The major differences in the different caries lesions focused on the relative spectral intensity range of 565-750 nm. The spectral parameter, defined as the ratio of wavebands at 565-750 nm to the whole spectral range, was calculated. The image component ratio R/(G + B) of color components was statistically computed by considering the spectral parameters (e.g. autofluorescence, optical filter, and spectral sensitivity) in our fluorescence color imaging system. Results showed that the spectral parameter and image component ratio presented a linear relation. Therefore, the image component ratio was graded as 1.62 to quantitatively classify sound, early decay, established decay, and severe decay tissues, respectively. Finally, the fluorescence images of caries were experimentally obtained, and the corresponding image component ratio distribution was compared with the classification result. A method to determine the numerical grades of caries using a fluorescence imaging system was proposed. This method can be applied to similar imaging systems.

  8. Fluorescence spectroscopy

    DEFF Research Database (Denmark)

    Bagatolli, Luis

    2016-01-01

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

  9. Separation and quantitation of phycobiliproteins using phytic acid in capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Viskari, Pertti J; Colyer, Christa L

    2002-10-01

    The similar electrophoretic mobilities and sizes of several of the phycobiliproteins, which are derived from the photosynthetic apparatus of cyanobacteria and eukaryotic algae, render their separation and quantitation a challenging problem. However, we have developed a suitable capillary electrophoresis (CE) method that employs a phytic acid-boric acid buffer and laser-induced fluorescence (LIF) detection with a single 594 nm He-Ne laser. This method takes advantage of the remarkably high quantum yields of these naturally fluorescent proteins, which can be attributed to their linear tetrapyrrole chromophores covalently bound to cysteinyl residues. As such, limits of detection of 1.18 x 10(-14), 5.26 x 10(-15), and 2.38 x 10(-15) mol/l were obtained for R-phycoerythrin, C-phycocyanin, and allophycocyanin proteins, respectively, with a linear dynamic range of eight orders of magnitude in each case. Unlike previously published CE-LIF methods, this work describes the separation of all three major classes of phycobiliproteins in under 5 min. Very good recoveries, ranging from 93.2 to 105.5%, were obtained for a standard mixture of the phycobiliproteins, based on seven-point calibration curves for both peak height and peak area. It is believed that this development will prove useful for the determination of phycobiliprotein content in naturally occurring cyanobacteria populations, thus providing a useful tool for understanding biological and chemical oceanographic processes.

  10. Development of an semi-automatic and sensitive photochemically induced fluorescence sensor for the determination of thiamethoxam in vegetables.

    Science.gov (United States)

    Jiménez-López, J; Ortega-Barrales, P; Ruiz-Medina, A

    2016-01-01

    The determination of thiamethoxam (TMX), a widely known neonicotinoid pesticide, by a multicommutated optosensing device implemented with photochemically induced fluorescence (PIF) has been developed. The combination of both methodologies allows, on one hand a quick on-line photodegradation of TMX and, on the other hand, the preconcentration, quantification and desorption of the fluorescent photoproduct generated once retained on C18 silica gel filling the flow-cell which was monitored at 353 and 407nm for excitation and emission wavelengths, respectively. The proposed analytical method presents a detection limit of 3.6ngmL(-1) by using Multicommutated Flow Injection Analysis (MCFIA) as flow methodology. Recovery experiments have been carried out in different kinds of vegetables at levels same or below the legislated maximum residue limit, demonstrating that this method combines advantages such as simplicity, high sensibility and high selectivity, in addition to fulfill the requirements for its applications in quality control. The obtained results in the analysis of real samples were in good agreement with those provided by a reference liquid chromatography (HPLC) method.

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

    Science.gov (United States)

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

    2015-12-01

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

  12. Evaluation of algorithm methods for fluorescence spectra of cancerous and normal human tissues

    Science.gov (United States)

    Pu, Yang; Wang, Wubao; Alfano, Robert R.

    2016-03-01

    The paper focus on the various algorithms on to unravel the fluorescence spectra by unmixing methods to identify cancerous and normal human tissues from the measured fluorescence spectroscopy. The biochemical or morphologic changes that cause fluorescence spectra variations would appear earlier than the histological approach; therefore, fluorescence spectroscopy holds a great promise as clinical tool for diagnosing early stage of carcinomas and other deceases for in vivo use. The method can further identify tissue biomarkers by decomposing the spectral contributions of different fluorescent molecules of interest. In this work, we investigate the performance of blind source un-mixing methods (backward model) and spectral fitting approaches (forward model) in decomposing the contributions of key fluorescent molecules from the tissue mixture background when certain selected excitation wavelength is applied. Pairs of adenocarcinoma as well as normal tissues confirmed by pathologist were excited by selective wavelength of 340 nm. The emission spectra of resected fresh tissue were used to evaluate the relative changes of collagen, reduced nicotinamide adenine dinucleotide (NADH), and Flavin by various spectral un-mixing methods. Two categories of algorithms: forward methods and Blind Source Separation [such as Principal Component Analysis (PCA) and Independent Component Analysis (ICA), and Nonnegative Matrix Factorization (NMF)] will be introduced and evaluated. The purpose of the spectral analysis is to discard the redundant information which conceals the difference between these two types of tissues, but keep their diagnostically significance. The facts predicted by different methods were compared to the gold standard of histopathology. The results indicate that these key fluorophores within tissue, e.g. tryptophan, collagen, and NADH, and flavin, show differences of relative contents of fluorophores among different types of human cancer and normal tissues. The

  13. A rapid and sensitive method to detect mycobacterium tuberculosis DNA by fluorescence polarization

    Institute of Scientific and Technical Information of China (English)

    白玉杰; 赵锦荣; 薛丽; 刘爱翔; 张文红; 郭晏海; 闫小君

    2004-01-01

    Objective: To develop a new high sensitivity, rapid and simple mycobacterium tuberculosis DNA detection method using fluorescence polarization technology. Methods: In our asymmetric PCR protocol, 100 times mole of TB-R primer was added than in the usual symmetric PCR to get enough single strands PCR product. The probe TB-5'-TAMRA and PCR products were mixed in a tube and incubate for 5 - 15 min at 46 C. The polarization (mp) was measured using victor2Multilabel Plate Reader. Results: Asymmetric and symmetric PCR products were analyzed by the FP method. Asymmetric PCR products are detected more sensitively than symmetric ones. The polarization values of probe associated with asymmetric products were much higher than with symmetric products. Conclusion: This fluorescence polarization assay in conjunction with asymmetric PCR is a powerful and widely applicable method for the rapid and sensitive detection of micro-organisms in clinical laboratories.

  14. Probing the connection of PBSs to the photosystems in Spirulina platensis by artificially induced fluorescence fluctuations

    Energy Technology Data Exchange (ETDEWEB)

    Li Heng [Key Laboratory of Photochemistry, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080 (China); Yang Shuzhen [Key Laboratory of Photochemistry, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080 (China); Xie Jie [Key Laboratory of Photochemistry, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080 (China); Zhao Jingquan [Key Laboratory of Photochemistry, Center for Molecular Science, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100080 (China)]. E-mail: zhaojq@iccas.ac.cn

    2007-01-15

    The molecular architecture and the structural connections of phycobilisomes (PBSs) to the photosystems in the intact cells of Spirulina platensis were studied by taking advantage of glycerol- and betaine-induced fluorescence fluctuations. Generally, with a selective excitation of C-phycocyanin (C-PC), glycerol could induce not only decoupling of PBSs from the photosystems but also of C-PC rods from allophycocyanin (APC) cores, while betaine could strengthen the connection of PBSs to the thylakoid membrane but induce a partial dissociation of PBS. On the other hand, glycerol did not exert an influence on the fluorescence spectra of the photosystems in isolated thylakoid membrane. Therefore, it was deduced that glycerol could provide a molecular environment to weaken the hydrophobic interactions of not only the L{sub CM} with the membrane but also the linker polypeptides to the water-soluble phycobiliproteins (C-PC and APC), while the betaine could strengthen the hydrophobic interaction of L{sub CM} with the membrane but weaken the electrostatic interaction of linker polypeptides to C-PC and/or APC.

  15. Choline-induced selective fluorescence quenching of acetylcholinesterase conjugated Au@BSA clusters.

    Science.gov (United States)

    Mathew, Meegle S; Baksi, Ananya; Pradeep, T; Joseph, Kuruvilla

    2016-07-15

    We have developed a highly selective sensitive fluorescent detection of acetylcholine (ACh) using bovine serum albumin (BSA) protected atomically precise clusters of gold. The gold quantum clusters (AuQC@BSA) synthesized using bovine serum albumin and conjugated with acetylcholinesterase (AChE), an enzyme specific for acetylcholine, resulting in AuQC@BSA-AChE. The enzyme, AChE hydrolyzes acetylcholine (ACh) to choline (Ch) which in turn interacts with AuQC@BSA-AChE and quenches its fluorescence, enabling sensing. We have carried out the real time monitoring of the hydrolysis of ACh using electrospray ionization mass spectrometry (ESI MS) to find out the mechanism of fluorescent quenching. The validity of present method for determination of concentration of acetylcholine in real system such as blood was demonstrated. Further, the sensor, AuQC@BSA-AChE can be easily coated on paper and an efficient and cheap sensor can be developed and detection limit for ACh is found to be 10nM. The fluorescent intensity of AuQC@BSA-AChE is sensitive towards acetylcholine in range of 10nM to 6.4µM. This suggests that AuQC@BSA-AChE has an excellent potential to be used for diagnosis of various neuropsychological and neuropsychiatric disorders.

  16. Hydrangea-like magneto-fluorescent nanoparticles through thiol-inducing assembly

    Science.gov (United States)

    Chen, Shun; Zhang, Junjun; Song, Shaokun; Xiong, Chuanxi; Dong, Lijie

    2017-01-01

    Magneto-fluorescent nanoparticles (NPs), recognized as an emerging class of materials, have drawn much attention because of their potential applications. Due to surface functionalization and thiol-metal bonds, a simple method has been put forward for fabricating hydrangea-like magneto-fluorescent Fe3O4-SH@QD NPs, through assembling thiol-modified Fe3O4 NPs with sub-size multi-layer core/shell CdSe/CdS/ZnS QDs. After a refined but controllable silane hydrolysis process, thiol-modified Fe3O4 was fabricated, resulting in Fe3O4-SH@QD NPs with QDs, while preventing the quenching of the QDs. As a result, the core Fe3O4 NPs were 18 nm in diameter, while the scattered CdSe/CdS/ZnS QDs were 7 nm in diameter. The resultant magneto-fluorescent Fe3O4-SH@QD NPs exhibit efficient fluorescence, superparamagnetism at room temperature, and rapid response to the external field, which make them ideal candidates for difunctional probes in MRI and bio-labels, targeting and photodynamic therapy, and cell tracking and separation.

  17. Temperature-Induced Protein Conformational Changes in Barley Root Plasma Membrane-Enriched Microsomes: II. Intrinsic Protein Fluorescence.

    Science.gov (United States)

    Caldwell, C R

    1987-07-01

    The membrane-bound proteins of barley (Hordeum vulgare L. cv Conquest) root plasma membrane-enriched microsomes displayed fluorescence typical of protein-associated trytophan residues. The protein fluorescence intensity was sensitive to variations in sample temperature. The temperature-induced decline in protein fluorescence intensity was nonlinear with slope discontinuities at about 12 and 32 degrees C. Detergents at levels above their critical micelle concentration enhanced protein fluorescence. Glutaraldehyde reduced protein fluorescence. Protein fluorescence polarization increased at temperatures above 30 degrees C. Both the rate of tryptophan photoionization and the fluorescence intensity of the photoionization products suggested alterations in membrane protein conformation between 12 and 32 degrees C. The quenching of the intrinsic protein fluorescence by acrylamide and potassium iodide indicated changes in accessibility of the extrinsic agents to the protein tryptophan residues beginning at about 14 degrees C. The results indicate thermally induced changes in the dynamics of the membrane proteins over the temperature range of 12 to 32 degrees C which could account for the complex temperature dependence of the barley root plasma membrane ATPase.

  18. Capillary electrophoresis with laser-induced fluorescence detection for studying amino acid uptake by yeast during beer fermentation.

    Science.gov (United States)

    Turkia, Heidi; Sirén, Heli; Penttilä, Merja; Pitkänen, Juha-Pekka

    2015-01-01

    The amino acid composition of cultivation broth is known to affect the biomass accumulation, productivity, and vitality of yeast during cultivation. A separation method based on capillary electrophoresis with laser-induced fluorescence (LIF) detection was developed for the determination of amino acid consumption by Saccharomyces cerevisiae during beer fermentation. Intraday relative standard deviations were less than 2.1% for migration times and between 2.9% and 9.9% for peak areas. Interday relative standard deviations were less than 2.5% for migration times and between 4.4% and 18.9% for peak areas. The quantification limit was even as low as 62.5 pM which equals to below attomole level detection. The method was applied to study the rate of amino acid utilization during beer fermentation.

  19. Trace Detection of Metastable Helium Molecules in Superfluid Helium by Laser-Induced Fluorescence

    CERN Document Server

    McKinsey, D N; Nikkel, J A; Rellergert, W

    2005-01-01

    We describe an approach to detecting ionizing radiation that combines the special properties of superfluid helium with the sensitivity of quantum optics techniques. Ionization in liquid helium results in the copious production of metastable helium molecules, which can be detected by laser-induced fluorescence. Each molecule can be probed many times using a cycling transition, resulting in the detection of individual molecules with high signal to noise. This technique could be used to detect neutrinos, weakly interacting massive particles, and ultracold neutrons, and to image superfluid flow in liquid He-4.

  20. Fluorescence Ratiometric Properties Induced by Nanoparticle Plasmonics and Nanoscale Dye Dynamics

    Directory of Open Access Journals (Sweden)

    Aron Hakonen

    2013-01-01

    Full Text Available Nanoscale transport of merocyanine 540 within/near the plasmon field of gold nanoparticles was recognized as an effective inducer of single-excitation dual-emission ratiometric properties. With a high concentration of the signal transducer (ammonium, a 700% increase in fluorescence was observed at the new red-shifted emission maximum, compared to a nanoparticle free sensor membrane. A previously nonrecognized isosbestic point is demonstrated at  nm. The mechanism can be utilized for enhanced and simplified ratiometric optical chemical sensors and potentially for thin film engineering to make solar cells more effective and stable by a broader and more regulated absorption.

  1. Theoretical study of determining orientation and alignment of symmetric top molecule using laser-induced fluorescence

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    General expressions used for extracting the orientation and alignment parameters of a symmetric top molecule from laser-induced fluorescence (LIF) intensity are derived by employing the density matrix approach. The molecular orientation and alignment are described by molecular state multipoles. Excitation and detection are circularly and linearly polarized lights, respectively. In general cases, the LIF intensity is a complex function of the initial molecular state multipoles, the dynamic factors and the excitation-detection geometrical factors. It contains a population, ten orientation and fourteen alignment multipoles. The problem of how to extract the initial molecular state multipoles from the resolved LIF intensity is discussed.

  2. Acetone, a laser-induced fluorescence study with rotational resolution at 320 nm

    Science.gov (United States)

    Zuckermann, Hanna; Haas, Yehuda; Drabbels, Marcel; Heinze, Johannes; Meerts, W. Leo; Reuss, Joerg; van Bladel, John

    1992-06-01

    The forbidden S 1←S 0 transition of acetone has been investigated by laser-induced fluorescence measurements with a resolution of 270 MHz. The rotational structure demonstrates, that (i) one deals with a-type transitions and (ii) there is a strong coupling between the torsional motion of the two CH 3 groups and the tunneling, out-of-plane wagging motion ( v23) of acetone. The interpretation of torsion-vibrational combination bands is less conclusive and thus the discussion still has a preliminary character.

  3. In-vivo optical detection of cancer using chlorin e6 – polyvinylpyrrolidone induced fluorescence imaging and spectroscopy

    OpenAIRE

    Soo Khee; Bhuvaneswari Ramaswamy; Thong Patricia SP; Chin William WL; Heng Paul WS; Olivo Malini

    2009-01-01

    Abstract Background Photosensitizer based fluorescence imaging and spectroscopy is fast becoming a promising approach for cancer detection. The purpose of this study was to examine the use of the photosensitizer chlorin e6 (Ce6) formulated in polyvinylpyrrolidone (PVP) as a potential exogenous fluorophore for fluorescence imaging and spectroscopic detection of human cancer tissue xenografted in preclinical models as well as in a patient. Methods Fluorescence imaging was performed on MGH human...

  4. ARTICLES: Laser-induced Fluorescence Spectrum of CoS Between 15200 and 19000 cm-1

    Science.gov (United States)

    Zhen, Jun-feng; Wang, Li; Qin, Cheng-bing; Zhang, Qun; Chen, Yang

    2010-06-01

    Laser-induced fluorescence excitation spectra of jet-cooled CoS molecules have been recorded in the energy range of 15200-19000 cm-1. Five transition progressions have been reported for the first time, the assignments of these progressions have been derived from a rotational analysis of vibronic bands and they are determined to be [15.58]4Δ7/2-X4Δ7/2, [16.02]4Δ7/2-X4Δ7/2, [16.50]4Δ7/2-X4Δ7/2, [17.80]4Π5/2-X4Δ7/2, and [18.00]4Δ7/2-X4Δ7/2 transitions. In addition, under the supersonic jet condition the fluorescent lifetimes of these vibronic states were measured by exponentially fitting the fluorescence decay. Based on the observed spectra and the measured lifetimes of the vibronic states, the newly identified electronic states are discussed.

  5. Laser Induced Fluorescence Spectroscopy of Neutral and Ionized Polycyclic Aromatic Hydrocarbons in the Cosmic Simulation Chamber

    Science.gov (United States)

    Bejaoui, Salma; Salama, Farid; Contreras, Cesar; Sciamma O'Brien, Ella; Foing, Bernard; Pascale, Ehrenfreund

    2015-01-01

    Polycyclic aromatic hydrocarbon (PAH) molecules are considered the best carriers to account for the ubiquitous infrared emission bands. PAHs have also been proposed as candidates to explain the diffuse interstellar bands (DIBs), a series of absorption features seen on the interstellar extinction curve and are plausible carriers for the extended red emission (ERE), a photoluminescent process associated with a wide variety of interstellar environments. Extensive efforts have been devoted over the past two decades to characterize the physical and chemical properties of PAH molecules and ions in space. Absorption spectra of PAH molecules and ions trapped in solid matrices have been compared to the DIBs. Absorption spectra of several cold, isolated gas-phase PAHs have also been measured under experimental conditions that mimic the interstellar conditions. The purpose of this study is to provide a new dimension to the existing spectroscopic database of neutral and single ionized PAHs that is largely based on absorption spectra by adding emission spectroscopy data. The measurements are based on the laser induced fluorescence (LIF) technique and are performed with the Pulsed Discharge Nozzle (PDN) of the COSmIC laboratory facility at NASA Ames laboratory. The PDN generates a plasma in a free supersonic jet expansion to simulate the physical and the chemical conditions in interstellar environments. We focus, here, on the fluorescence spectra of large neutral PAHs and their cations where there is a lack of fluorescence spectroscopy data. The astronomical implications of the data (e.g., ERE) are examined.

  6. Laser-induced fluorescence detection platform for point-of-care testing

    Science.gov (United States)

    Berner, Marcel; Hilbig, Urs; Schubert, Markus B.; Gauglitz, Günter

    2017-08-01

    Point-of-care testing (POCT) devices for continuous low-cost monitoring of critical patient parameters require miniaturized and integrated setups for performing quick high-sensitivity analyses, away from central clinical laboratories. This work presents a novel and promising laser-induced fluorescence platform for measurements in direct optical test formats that leads towards such powerful POCT devices based on fluorescence-labeled immunoassays. Ultimate sensitivity of thin film photodetectors, integrated with microfluidics, and a comprehensive optimization of all system components aim at low-level signal detection in the targeted biosensor application. The setup acquires fluorescence signals from the volume of a microfluidic channel. An innovative sandwiching process forms a flow channel in the microfluidic chips by embedding laser-cut double-sided adhesive tapes. The custom fit of amorphous silicon based photodiode arrays to the geometry of the flow channel enables miniaturization, fully adequate for POCT devices. A free-beam laser excitation with line focus provides excellent alignment stability, allows for easy and reliable swapping of the disposable microfluidic chips, and therewith greatly improves the ease of use of the resulting integrated device. As a proof-of-concept of this novel in-volume measurement approach, the limit of detection for the dye DY636-COOH in pure water as a model fluorophore is examined and found to be 26 nmol l-1 .

  7. Mechanism of laser induced fluorescence signal generation in InCl3-ethanol mixture flames

    Science.gov (United States)

    Fang, Bolang; Hu, Zhiyun; Zhang, Zhenrong; Li, Guohua; Shao, Jun; Feng, Guobin

    2017-05-01

    Nonlinear regime Two-line Atomic Fluorescence (NTLAF) is a promising technique for two-dimensional thermometry. A key challenge is seeding of indium atoms into flame. This work aims at investigating the mechanism of Indium LIF signal generation in a fuel-rich InCl3-ethanol premixed flame. Several types of images including natural emission of the flame itself, natural emission of CH, natural emission of OH, natural emission at 410 nm/451 nm of indium atom, and laser induced fluorescence at 410 nm/451 nm were obtained. The indium atom was generated in the flame front, and could survive in the post-flame zone for a while which is benefit for making NTLAF measurements. Further detail mechanism of fluorescence signals generation in InCl3-ethanol solution burning was investigated. The conclusion which probable to be drew is that to gain high NTLAF signals, the size of liquid droplets should be well controlled, neither to be too large nor to be gasified.

  8. An Intelligent Optical Dissolved Oxygen Measurement Method Based on a Fluorescent Quenching Mechanism

    Directory of Open Access Journals (Sweden)

    Fengmei Li

    2015-12-01

    Full Text Available Dissolved oxygen (DO is a key factor that influences the healthy growth of fishes in aquaculture. The DO content changes with the aquatic environment and should therefore be monitored online. However, traditional measurement methods, such as iodometry and other chemical analysis methods, are not suitable for online monitoring. The Clark method is not stable enough for extended periods of monitoring. To solve these problems, this paper proposes an intelligent DO measurement method based on the fluorescence quenching mechanism. The measurement system is composed of fluorescent quenching detection, signal conditioning, intelligent processing, and power supply modules. The optical probe adopts the fluorescent quenching mechanism to detect the DO content and solves the problem, whereas traditional chemical methods are easily influenced by the environment. The optical probe contains a thermistor and dual excitation sources to isolate visible parasitic light and execute a compensation strategy. The intelligent processing module adopts the IEEE 1451.2 standard and realizes intelligent compensation. Experimental results show that the optical measurement method is stable, accurate, and suitable for online DO monitoring in aquaculture applications.

  9. Ns-scale time-resolved laser induced fluorescence imaging for detection of fecal contamination on apples

    Science.gov (United States)

    Kim, Moon S.; Lefcourt, Alan M.; Chen, Yud-Ren

    2004-11-01

    Our laboratory has been utilizing fluorescence techniques as a potential means for detection of quality and wholesomeness of food products. A system with a short pulse light source such as a laser coupled with a gated detector can be used to harvest fluorescence in ambient light conditions from biological samples with relatively low fluorescence yields. We present a versatile multispectral laser-induced fluorescence (LIF) imaging system capable of ns-scale time resolved fluorescence. The system is equipped with a tunable pulse laser system that emits in the visible range from 410 nm to 690 nm. Ns-scale, time-dependent multispectral fluorescence emissions of apples contaminated with a range of diluted cow feces were acquired. Four spectral bands, F670, F680, F685 and F730, centered near the emission peak wavelengths of the major constituents responsible for the red fluorescence emissions from apples artificially contaminated with cow feces were examined to determine a suitable single red fluorescence band and optimal ns-gate window for detection of fecal contamination on apples. The results based on the ns decay curves showed that 670 nm with 10 nm full width at half maximum (FWHM) at a gate-delay of 4 ns from the laser excitation peak provided the greatest differences in time-dependent fluorescence responses between feces contaminated spots and apples surfaces.

  10. A fluorescence method to detect and quantitate sterol esterification by lecithin:cholesterol acyltransferase.

    Science.gov (United States)

    Homan, Reynold; Esmaeil, Nadia; Mendelsohn, Laurel; Kato, Gregory J

    2013-10-01

    We describe a simple but sensitive fluorescence method to accurately detect the esterification activity of lecithin:cholesterol acyltransferase (LCAT). The new assay protocol employs a convenient mix, incubate, and measure scheme. This is possible by using the fluorescent sterol dehydroergosterol (DHE) in place of cholesterol as the LCAT substrate. The assay method is further enhanced by incorporation of an amphiphilic peptide in place of apolipoprotein A-I as the lipid emulsifier and LCAT activator. Specific fluorescence detection of DHE ester synthesis is achieved by employing cholesterol oxidase to selectively render unesterified DHE nonfluorescent. The assay accurately detects LCAT activity in buffer and in plasma that is depleted of apolipoprotein B lipoproteins by selective precipitation. Analysis of LCAT activity in plasmas from control subjects and sickle cell disease (SCD) patients confirms previous reports of reduced LCAT activity in SCD and demonstrates a strong correlation between plasma LCAT activity and LCAT content. The fluorescent assay combines the sensitivity of radiochemical assays with the simplicity of nonradiochemical assays to obtain accurate and robust measurement of LCAT esterification activity. Copyright © 2013 Elsevier Inc. All rights reserved.

  11. Research on the discrimination methods of algae based on the fluorescence excitation spectra

    Institute of Scientific and Technical Information of China (English)

    HU Xupeng; SU Rongguo; ZOU Weiming; REN Shijun; WANG Hongtao; CHAI Xiaoping; WANG Yiming

    2010-01-01

    The excitation spectra of chlorophyll(Chl)fluorescence can be used to differentiate phytoplankton populations at phylum level in vivo and in situ within a few minutes.The investigated phytoplankton divisions(Dinophyta,Bacillariophyta,Chrysophyta,Cyanophyta,Cryptophyta,Chlorophyta)are each characterized by a specific composition of photosynthetic antenna pigments and,consequently,by a specific excitation spectrum of the Chl fluorescence.Norm excitation spectra(emission of 680 nm and excitation of 400-600 nm)of every division were obtained from several species per division by a F4500 fluorescence spectrophotometer.Fisher's linear discriminant analysis of the norm spectra shows that the divisions could be discriminated.The discrimination method,established by multivariate linear regression and weighted least squares,was used to differentiate the phytoplankton samples cultured in the laboratory and samples collected from the Jiaozhao Bay at division level.The correctly discriminated samples were more than 94% for single algal species ones,more than 84% for simulatively mixed ones,more than 83% for real mixed ones and 100% for samples collected from the Jiaozhou Bay for the dominant species.The method for phytoplankton differentiation described here can be applied to routine checking by fluorescence spectrophotometer,and benefit the monitoring and supervision tasks related to phytoplankton populations in the marine environments.

  12. A Graphene Oxide-Based Fluorescent Method for the Detection of Human Chorionic Gonadotropin

    Directory of Open Access Journals (Sweden)

    Ning Xia

    2016-10-01

    Full Text Available Human chorionic gonadotropin (hCG has been regarded as a biomarker for the diagnosis of pregnancy and some cancers. Because the currently used methods (e.g., disposable Point of Care Testing (POCT device for hCG detection require the use of many less stable antibodies, simple and cost-effective methods for the sensitive and selective detection of hCG have always been desired. In this work, we have developed a graphene oxide (GO-based fluorescent platform for the detection of hCG using a fluorescein isothiocyanate (FITC-labeled hCG-specific binding peptide aptamer (denoted as FITC-PPLRINRHILTR as the probe, which can be manufactured cheaply and consistently. Specifically, FITC-PPLRINRHILTR adsorbed onto the surface of GO via electrostatic interaction showed a poor fluorescence signal. The specific binding of hCG to FITC-PPLRINRHILTR resulted in the release of the peptide from the GO surface. As a result, an enhanced fluorescence signal was observed. The fluorescence intensity was directly proportional to the hCG concentration in the range of 0.05–20 IU/mL. The detection limit was found to be 20 mIU/mL. The amenability of the strategy to hCG analysis in biological fluids was demonstrated by assaying hCG in the urine samples.

  13. A general method for the detection of large CAG repeat expansions by fluorescent PCR.

    Science.gov (United States)

    Warner, J P; Barron, L H; Goudie, D; Kelly, K; Dow, D; Fitzpatrick, D R; Brock, D J

    1996-12-01

    The expansion of a tandemly repeated trinucleotide sequence, CAG, is the mutational mechanism for several human genetic diseases. We present a generally applicable PCR amplification method using a fluorescently labelled locus specific primer flanking the CAG repeat together with paired primers amplifying from multiple priming sites within the CAG repeat. Triplet repeat primed PCR (TP PCR) gives a characteristic ladder on the fluorescence trace enabling the rapid identification of large pathogenetic CAG repeats that cannot be amplified using flanking primers. We used our method to test a cohort of 183 people from myotonic dystrophy families including unaffected subjects and spouses. Eighty five clinically affected subjects with expanded alleles on Southern blot analysis were all correctly identified by TP PCR. This method is applicable for any human diseases involving CAG repeat expansions.

  14. 10 CFR Appendix Q to Subpart B of... - Uniform Test Method for Measuring the Energy Consumption of Fluorescent Lamp Ballasts

    Science.gov (United States)

    2010-01-01

    ... 10 Energy 3 2010-01-01 2010-01-01 false Uniform Test Method for Measuring the Energy Consumption... Appendix Q to Subpart B of Part 430—Uniform Test Method for Measuring the Energy Consumption of Fluorescent... reference; see § 430.3). The test for measuring standby mode energy consumption of fluorescent lamp ballasts...

  15. Silica nanoparticle-based microfluidic immunosensor with laser-induced fluorescence detection for the quantification of immunoreactive trypsin.

    Science.gov (United States)

    Seia, Marco A; Stege, Patricia W; Pereira, Sirley V; De Vito, Irma E; Raba, Julio; Messina, Germán A

    2014-10-15

    The purpose of this study was to develop a silica nanoparticle-based immunosensor with laser-induced fluorescence (LIF) as a detection system. The proposed device was applied to quantify the immunoreactive trypsin (IRT) in cystic fibrosis (CF) newborn screening. A new ultrasonic procedure was used to extract the IRT from blood spot samples collected on filter papers. After extraction, the IRT reacted immunologically with anti-IRT monoclonal antibodies immobilized on a microfluidic glass chip modified with 3-aminopropyl functionalized silica nanoparticles (APSN-APTES-modified glass chips). The bounded IRT was quantified by horseradish peroxidase (HRP)-conjugated anti-IRT antibody (anti-IRT-Ab) using 10-acetyl-3,7-dihydroxyphenoxazine (ADHP) as enzymatic mediator. The HRP catalyzed the oxidation of nonfluorescent ADHP to highly fluorescent resorufin, which was measured by LIF detector, using excitation lambda at 561nm and emission at 585nm. The detection limits (LODs) calculated for LIF detection and for a commercial enzyme-linked immunosorbent assay (ELISA) test kit were 0.87 and 4.2ngml(-1), respectively. The within- and between-assay variation coefficients for the LIF detection procedure were below 6.5%. The blood spot samples collected on filter papers were analyzed with the proposed method, and the results were compared with those of the reference ELISA method, demonstrating a potential usefulness for the clinical assessment of IRT during the early neonatal period.

  16. The analysis of fountain pen inks by capillary electrophoresis with ultraviolet/visible absorbance and laser-induced fluorescence detection.

    Science.gov (United States)

    Rohde, E; Vogt, C; Heineman, W R

    1998-01-01

    Information on the identity of inks adds to the circumstantial evidence in legal cases involving fraudulent documents. In combination with optical methods, multiple thin-layer chromatography (TLC) is currently the analytical tool used by forensic chemists to separate, compare and distinguish inks based on their dye composition. In our studies, capillary electrophoresis (CE) was used for the analysis of water-soluble fountain pen inks. Inks are complex mixtures of synthetic organic and inorganic dyes, surfactants, resins and other components. The investigations included the development of an electrophoretic separation method, the optimization of an extraction procedure for inks from paper as well as the evaluation of ultraviolet/visible (UV/VIS) absorbance and laser-induced fluorescence (LIF) detection for the analysis of inks by CE. Good results for the separation of 17 blue and black inks of different manufacturers and countries of origin were obtained with 100 mM borate buffer, pH 8.0, containing 20% methanol. The electropherograms of the inks and their extracts from paper showed patterns that were in most cases distinctly different from each other. Ultraviolet/visible scans can be used to compare spectra of the separated main and trace components of inks. Fluorescence detection at different excitation and emission wavelengths was more sensitive, but added to the complexity of the electropherograms due to the excitation of coextracted fluorescing paper components. The resolution power of CE combined with the information content provided by the detection modes investigated prove CE to be a powerful tool for the identification of water-soluble inks used on paper documents.

  17. SiO multi-line laser-induced fluorescence for quantitative temperature imaging in flame-synthesis of nanoparticles

    Science.gov (United States)

    Chrystie, Robin S. M.; Feroughi, Omid M.; Dreier, Thomas; Schulz, Christof

    2017-04-01

    Silicon monoxide (SiO) is an intermediate in the gas-phase synthesis of SiO2 nanoparticles and coatings. We demonstrate a method for in situ imaging the gas-phase temperature via multi-line laser-induced fluorescence (LIF) using excitation in the A 1Π- X 1Σ+(0,0) band near 235 nm. A low-pressure lean (3 kPa, φ = 0.39) premixed hydrogen/oxygen flame was seeded with 210 ppm hexamethyldisiloxane (HMDSO) to produce SiO2 nanoparticles. Spectral regions with no interference from other species in the flame were located, and the excitation-spectral range that provides the best temperature sensitivity was determined from numerical experiments. Quenching rates of the selected transitions were also determined from fluorescence lifetime measurements, and found to be independent of the excited rotational state. Upon laser light-sheet excitation, images of fluorescence were recorded for a sequence of excitation wavelengths and pixel-wise multi-line fitting of the spectra yields temperature images. The results were compared against multi-line NO-LIF temperature imaging measurements using the A 2Σ+- X 2Π(0,0) band near 225 nm from 500 ppm NO added to the gas flow as a thermometry target. Both methods show good qualitative agreement with each other and demonstrate that temperature can be evaluated from the zone in the reactor where SiO is naturally present without adding tracers. SiO LIF exhibited high signal-to-noise ratios of the order of ten times that of NO LIF.

  18. Quantitative measurement of hydroxyl radical (OH) concentration in premixed flat flame by combining laser-induced fluorescence and direct absorption spectroscopy

    Science.gov (United States)

    Chen, Shuang; Su, Tie; Li, Zhong-Shan; Bai, Han-Chen; Yan, Bo; Yang, Fu-Rong

    2016-10-01

    An accurate and reasonable technique combining direct absorption spectroscopy and laser-induced fluorescence (LIF) methods is developed to quantitatively measure the concentrations of hydroxyl in CH4/air flat laminar flame. In our approach, particular attention is paid to the linear laser-induced fluorescence and absorption processes, and experimental details as well. Through measuring the temperature, LIF signal distribution and integrated absorption, spatially absolute OH concentrations profiles are successfully resolved. These experimental results are then compared with the numerical simulation. It is proved that the good quality of the results implies that this method is suitable for calibrating the OH-PLIF measurement in a practical combustor. Project supported by the National Natural Science Foundation of China (Grant No. 11272338), the Science and Technology on Scramjet Key Laboratory Funding, China (Grant No. STSKFKT 2013004), and the China Scholarship Council.

  19. Suppression of Kasha's rule as a mechanism for fluorescent molecular rotors and aggregation-induced emission

    Science.gov (United States)

    Qian, Hai; Cousins, Morgan E.; Horak, Erik H.; Wakefield, Audrey; Liptak, Matthew D.; Aprahamian, Ivan

    2017-01-01

    Although there are some proposed explanations for aggregation-induced emission, a phenomenon with applications that range from biosensors to organic light-emitting diodes, current understanding of the quantum-mechanical origin of this photophysical behaviour is limited. To address this issue, we assessed the emission properties of a series of BF2-hydrazone-based dyes as a function of solvent viscosity. These molecules turned out to be highly efficient fluorescent molecular rotors. This property, in addition to them being aggregation-induced emission luminogens, enabled us to probe deeper into their emission mechanism. Time-dependent density functional theory calculations and experimental results showed that the emission is not from the S1 state, as predicted from Kasha's rule, but from a higher energy (>S1) state. Furthermore, we found that suppression of internal conversion to the dark S1 state by restricting the rotor rotation enhances fluorescence, which leads to the proposal that suppression of Kasha's rule is the photophysical mechanism responsible for emission in both viscous solution and the solid state.

  20. PNA-induced assembly of fluorescent proteins using DNA as a framework.

    Science.gov (United States)

    Gholami, Zahra; Brunsveld, Luc; Hanley, Quentin

    2013-08-21

    Controlled alignment of proteins on molecular frameworks requires the development of facile and orthogonal chemical approaches and molecular scaffolds. In this work, protein-PNA conjugates are brought forward as new chemical components allowing efficient assembly and alignment on DNA scaffolds. Site-selective monomeric teal fluorescent protein (mTFP)-peptide nucleic acid (PNA) (mTFP-PNA) conjugation was achieved by covalent linkage of the PNA to the protein through expressed protein ligation (EPL). A DNA beacon, with 6-Fam and Dabcyl at its ends, acts as a framework to create an assembled hetero-FRET system with the mTFP-PNA conjugate. Using fluorescence intensity, frequency domain lifetime measurements, and anisotropy measurements, the system was shown to produce FRET as indicated by decreased donor intensity, decreased donor lifetime, and increased donor anisotropy. Extension of the DNA scaffold allowed for the assembly of multiple mTFP-PNA constructs. Efficient formation of protein dimers and oligomers on the DNA-PNA frameworks could be shown, as visualized via size exclusion chromatography (SEC) and electrophoresis (SDS-PAGE). Assembly of multiple proteins in a row induced homo-FRET for the mTFP-PNA's assembled on the DNA scaffolds. The oligonucleotide framework allows an induced and controllable assembly of proteins by fusing them to PNAs directed to align on DNA scaffolds.

  1. A fluorescence anisotropy method for measuring protein concentration in complex cell culture media.

    Science.gov (United States)

    Groza, Radu Constantin; Calvet, Amandine; Ryder, Alan G

    2014-04-22

    The rapid, quantitative analysis of the complex cell culture media used in biopharmaceutical manufacturing is of critical importance. Requirements for cell culture media composition profiling, or changes in specific analyte concentrations (e.g. amino acids in the media or product protein in the bioprocess broth) often necessitate the use of complicated analytical methods and extensive sample handling. Rapid spectroscopic methods like multi-dimensional fluorescence (MDF) spectroscopy have been successfully applied for the routine determination of compositional changes in cell culture media and bioprocess broths. Quantifying macromolecules in cell culture media is a specific challenge as there is a need to implement measurements rapidly on the prepared media. However, the use of standard fluorescence spectroscopy is complicated by the emission overlap from many media components. Here, we demonstrate how combining anisotropy measurements with standard total synchronous fluorescence spectroscopy (TSFS) provides a rapid, accurate quantitation method for cell culture media. Anisotropy provides emission resolution between large and small fluorophores while TSFS provides a robust measurement space. Model cell culture media was prepared using yeastolate (2.5 mg mL(-1)) spiked with bovine serum albumin (0 to 5 mg mL(-1)). Using this method, protein emission is clearly discriminated from background yeastolate emission, allowing for accurate bovine serum albumin (BSA) quantification over a 0.1 to 4.0 mg mL(-1) range with a limit of detection (LOD) of 13.8 μg mL(-1). Copyright © 2014. Published by Elsevier B.V.

  2. Determination of human albumin in serum and urine samples by constant-energy synchronous fluorescence method.

    Science.gov (United States)

    Madrakian, Tayyebeh; Bagheri, Habibollah; Afkhami, Abbas

    2015-08-01

    A sensitive spectrofluorimetric method using constant-energy synchronous fluorescence technique is proposed for the determination of human albumin without separation. In this method, no reagent was used for enhancement of the fluorescence signal of albumin in the solution. Effects of some parameters, such as energy difference between excitation and emission monochromators (ΔE), emission and excitation slit widths and scan rate of wavelength were studied and the optimum conditions were established. For this purpose factorial design and response surface method were employed for optimization of the effective parameters on the fluorescence signal. The results showed that the scan rate of the wavelength has no significant effect on the analytical signal. The calibration curve was linear in the range 0.1-220.0 µg mL(-1) of albumin with a detection limit of 7.0 × 10(-3)  µg mL(-1). The relative standard deviations (RSD) for six replicate measurements of albumin were calculated as 2.2%, 1.7% and 1.3% for 0.5, 10.0 and 100.0 µg mL(-1) albumin, respectively. Furthermore the proposed method has been employed for the determination of albumin in human serum and urine samples.

  3. A method of measuring gold nanoparticle concentrations by x-ray fluorescence for biomedical applications

    Energy Technology Data Exchange (ETDEWEB)

    Wu Di; Li Yuhua; Wong, Molly D.; Liu Hong [Center for Bioengineering and School of Electrical and Computer Engineering, University of Oklahoma, Norman, Oklahoma 73019 (United States)

    2013-05-15

    Purpose: This paper reports a technique that enables the quantitative determination of the concentration of gold nanoparticles (GNPs) through the accurate detection of their fluorescence radiation in the diagnostic x-ray spectrum. Methods: Experimentally, x-ray fluorescence spectra of 1.9 and 15 nm GNP solutions are measured using an x-ray spectrometer, individually and within chicken breast tissue samples. An optimal combination of excitation and emission filters is determined to segregate the fluorescence spectra at 66.99 and 68.80 keV from the background scattering. A roadmap method is developed that subtracts the scattered radiation (acquired before the insertion of GNP solutions) from the signal radiation acquired after the GNP solutions are inserted. Results: The methods effectively minimize the background scattering in the spectrum measurements, showing linear relationships between GNP solutions from 0.1% to 10% weight concentration and from 0.1% to 1.0% weight concentration inside a chicken breast tissue sample. Conclusions: The investigation demonstrated the potential of imaging gold nanoparticles quantitatively in vivo for in-tissue studies, but future studies will be needed to investigate the ability to apply this method to clinical applications.

  4. Comparison of three-dimensional fluorescence analysis methods for predicting formation of trihalomethanes and haloacetic acids

    Institute of Scientific and Technical Information of China (English)

    Nicolás M.Peleato; Robert C.Andrews

    2015-01-01

    This work investigated the application of several fluorescence excitation-emission matrix analysis methods as natural organic matter (NOM) indicators for use in predicting the formation of trihalomethanes (THMs) and haloacetic acids (HAAs).Waters from four different sources (two rivers and two lakes) were subjected to jar testing followed by 24 hr disinfection by-product formation tests using chlorine.NOM was quantified using three common measures:dissolved organic carbon,ultraviolet absorbance at 254 nm,and specific ultraviolet absorbance as well as by principal component analysis,peak picking,and parallel factor analysis of fluorescence spectra.Based on multi-linear modeling of THMs and HAAs,principle component (PC) scores resulted in the lowest mean squared prediction error of cross-folded test sets (THMs:43.7 (μg/L)2,HAAs:233.3 (μg/L)2).Inclusion of principle components representative of protein-like material significantly decreased prediction error for both THMs and HAAs.Parallel factor analysis did not identify a protein-like component and resulted in prediction errors similar to traditional NOM surrogates as well as fluorescence peak picking.These results support the value of fluorescence excitation-emission matrix-principal component analysis as a suitable NOM indicator in predicting the formation of THMs and HAAs for the water sources studied.

  5. Experimental Study of Laser-induced Fluorescence of Anthracene in Soil%土壤中蒽的激光诱导荧光实验研究

    Institute of Scientific and Technical Information of China (English)

    何俊; 邓琥; 武志翔; 马有良

    2011-01-01

    It is complicated to detect Polycyclic Aromatic Hydrocarbons (PAHs) contamination soil by High Performance Liquid Chromatography (HPLC) with the conventional method, the Laser-induced Fluorescence (LIF) is used to on-site detect anthracene, one of the PAHs in soil, and it is shown that the Laser-induced Fluorescence can facilitate the monitoring of the soil pollution. According to the fluorescence properties of anthracene, the optical path and sample holder are designed to directly excite the fluorescence of the mixture of anthracene and the soil sample by using Nd: YAG laser at 355 nm, and the fluorescence spectra are recorded by the fiber spectrometer. It is shown that there are obvious emission fluorescence peaks at 375 nm, 391 nm, 419 nm and 445 nm, and different mass firactions exhibit good linearity to the fluorescence intensities with the correlation coefficients up to 0.999. The effects of the changes in experimental conditions on the fluorescence properties of anthracene are studied, and it is demonstrated by experiment that it is feasible to on-site quantitatively detect anthracene in the soil by using Laser-induced Fluorescence.%常规利用液相色谱法检测土壤中的多环芳烃污染需要复杂的萃取过程,采用激光诱导荧光法直接检测土壤中的多环芳烃,有利于实现对土壤污染的监控.文章以多环芳烃中的蒽为研究对象,根据蒽的荧光特性,设计了实验光路和样品台,采用Nd:YAG激光器355 nm波长直接激发蒽和土壤混合样本的荧光,利用光纤光谱仪记录荧光光谱,实验发现在375nm、391nm、419nm和445nm出现了明显的发射荧光峰,不同质量分数和其荧光强度呈现出良好的线性,相关系数达到0.999;通过实验研究了实验条件的改变对土壤中蒽的荧光特性的影响.实验结果表明,利用激光诱导荧光的方法对土壤中蒽进行直接定量检测是可行的.

  6. Serum protein profile study of clinical samples using high performance liquid chromatography-laser induced fluorescence: case of cervical and oral cancers

    Science.gov (United States)

    Karemore, Gopal; Sujatha, .; Rai, Lavanya; Pai, Keerthilatha M.; Kartha, V. B.; Santhosh C., .

    2009-02-01

    The serum protein profiles of normal subjects, patients diagnosed with cervical cancer, and oral cancer were recorded using High Performance Liquid Chromatography combined with Laser Induced Fluorescence detection (HPLC-LIF). Serum protein profiles of the above three classes were tested for establishing the ability of HPLC-LIF protein profiling technique for discrimination, using hard clustering and Fuzzy clustering methods. The clustering algorithms have quite successfully classified the profiles as belonging to normal, cancer of cervix, and oral cancer conditions.

  7. One for all--a highly efficient and versatile method for fluorescent immunostaining in fish embryos.

    Directory of Open Access Journals (Sweden)

    Daigo Inoue

    Full Text Available BACKGROUND: For the detection and sub-cellular (co-localization of proteins in the context of the tissue or organism immunostaining in whole mount preparations or on sections is still the best approach. So far, each antibody required its own fixation and antigen retrieval protocol so that optimizing immunostaining turned out to be tedious and time consuming. METHODOLOGY/PRINCIPAL FINDING: Here we present a novel method to efficiently retrieve the antigen in a widely applicable standard protocol, facilitating fluorescent immunostaining of both cryosections and whole mount preparations in zebrafish (Danio rerio and medaka (Oryzias latipes. CONCLUSIONS/SIGNIFICANCE: Our method overcomes the loss of sections and damage of tissue and cell morphology, and allows parallel immunostaining in multiple colors, co-immunostaining with fluorescent proteins in transgenic fish lines and in combination with whole mount in situ hybridization.

  8. Modified in situ antimicrobial susceptibility testing method based on cyanobacteria chlorophyll a fluorescence.

    Science.gov (United States)

    Heliopoulos, Nikolaos S; Galeou, Angeliki; Papageorgiou, Sergios K; Favvas, Evangelos P; Katsaros, Fotios K; Stamatakis, Kostas

    2016-02-01

    The chlorophyll a fluorescence based antimicrobial susceptibility testing (AST) method presented in a previous work was based on the measurement of Chl a fluorescence of the gram(-) cyanobacterium Synechococcus sp. PCC 7942. Synechococcus sp. PCC 7942 as a gram(-) bacterium is affected by antibacterial agents via mechanisms affecting all gram(-) bacteria, however, as an exclusively phototrophic organism it would also be affected by photosynthesis inhibitory action of an agent that otherwise has no antibacterial properties. In this report, the method is modified by replacing the exclusively phototrophic Synechococcus sp. PCC 7942 with the Synechocystis sp. PCC 6714, capable of both phototrophic and heterotrophic growth in order to add versatility and better reflect the antibacterial effects of surfaces under study towards nonphotosynthetic bacteria. Copyright © 2015 Elsevier B.V. All rights reserved.

  9. Phase Structures of Microemulsions Determined by the Steady-State Fluorescence Method

    Institute of Scientific and Technical Information of China (English)

    2001-01-01

    The steady-state fluorescence method has been tentatively used to determine the phase structures of microemulsion systems consisting of cetyltrimethylammonium bromide (CTAB), n-butanol (n-C4H9OH), octane (n-C5H18), and water. The excimer/monomer intensity ratio (Ie/Im) of pyrene has demonstrated that the various structures in the microemulsion phase region can be distinguished. The results are consistent with electrical conductivity data already reported.

  10. An ultrasensitive method for the determination of melamine using cadmium telluride quantum dots as fluorescence probes

    Energy Technology Data Exchange (ETDEWEB)

    Li, Xiafei; Li, Jin; Kuang, Huiyan; Feng, Lei; Yi, Shoujun; Xia, Xiaodong; Huang, Haowen [School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China); Key Laboratory of Theoretical Chemistry and Molecular Simulation of Ministry of Education of China, Hunan University of Science and Technology, Xiangtan 411201 (China); Chen, Yong; Tang, Chunran [School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China); Zeng, Yunlong, E-mail: yunlongzeng1955@126.com [School of Chemistry and Chemical Engineering, Hunan University of Science and Technology, Xiangtan 411201 (China); Key Laboratory of Theoretical Chemistry and Molecular Simulation of Ministry of Education of China, Hunan University of Science and Technology, Xiangtan 411201 (China); State Key Laboratory for Chemo/Biosensing and Chemometrics, College of Chemistry and Chemical Engineering, Hunan University, Changsha 410082 (China)

    2013-11-13

    Graphical abstract: Melamine takes place of the TGA on the surface of TGA-CdTe QDs with negative charge to form melamine coated QDs changing the surface charge of the QDs, resulting the fluorescence quenched as the QDs aggregation occurred by electrostatic attraction of the two opposite charged nanocrystals. -- Highlights: •An ultrasensitive and selective method for the determination of melamine was developed at pH 11.0. •The selectivity of the method was improved. •The sensitivity of the method enhanced obviously as the CdTe QDs have higher QYs at pH 11. •The sensitivity and linear range for the analysis are size dependent using QDs PL probes. •Melamine takes the place of TGA resulting fluorescence quenched of QDs. -- Abstract: An ultrasensitive and simple method for the determination of melamine was developed based on the fluorescence quenching of thioglycolic acid (TGA) capped CdTe quantum dots (QDs) at pH 11.0. In strong alkaline aqueous solution, the selectivity of the method has been greatly improved due to most heavy metal ions show no interference as they are in the precipitation form or in their anion form. Furthermore, CdTe quantum dots have higher quantum yields at higher pH. The method has a wider concentration range and lower detection limit. The influence factors on the determination of melamine were investigated and the optimum conditions were determined. Under optimum conditions, the fluorescence intensity change of TGA coated CdTe quantum dots was linearly proportional to melamine over a concentration range from 1.0 × 10{sup −11} to 1.0 × 10{sup −5} mol L{sup −1} with a correlation coefficient of 0.9943 and a detection limit of 5 × 10{sup −12} mol L{sup −1}. The mechanism of fluorescence quenching of the QDs has been proposed based on the infrared spectroscopy information and electrophoresis experiments in presence of melamine under alkaline condition. The proposed method was employed to detect trace melamine in milk powder

  11. Uncertainties in hot-wire measurements of compressible turbulent flows implied by comparisons with laser-induced fluorescence

    Science.gov (United States)

    Mckenzie, R. L.; Logan, P.

    1986-01-01

    A hot-wire anemometer and a new nonintrusive laser-induced fluorescence (LIF) technique are used to survey a Mach 2 turbulent boundary layer. The hot-wire anemometer's ability to accurately measure mass flux, temperature, and density fluctuations in a compressible flow is examined by comparing its results with those obtained using LIF. Several methods of hot-wire calibration are used, and the uncertainties in their measurements of various fluctuating flow parameters are determined. The results show that although a hot-wire operated at high overheat can measure mass flux fluctuations, temperature and density fluctuations are not determined accurately from such measurements. However, a hot-wire operated at multiple overheats can be used to measure static and total temperature fluctuations. The presence of pressure fluctuations and their correlation with density can prevent the use of hot-wire data to determine density fluctuations.

  12. Decoding of quantum dots encoded microbeads using a hyperspectral fluorescence imaging method.

    Science.gov (United States)

    Liu, Yixi; Liu, Le; He, Yonghong; Zhu, Liang; Ma, Hui

    2015-05-19

    We presented a decoding method of quantum dots encoded microbeads with its fluorescence spectra using line scan hyperspectral fluorescence imaging (HFI) method. A HFI method was developed to attain both the spectra of fluorescence signal and the spatial information of the encoded microbeads. A decoding scheme was adopted to decode the spectra of multicolor microbeads acquired by the HFI system. Comparison experiments between the HFI system and the flow cytometer were conducted. The results showed that the HFI system has higher spectrum resolution; thus, more channels in spectral dimension can be used. The HFI system detection and decoding experiment with the single-stranded DNA (ssDNA) immobilized multicolor beads was done, and the result showed the efficiency of the HFI system. Surface modification of the microbeads by use of the polydopamine was characterized by the scanning electron microscopy and ssDNA immobilization was characterized by the laser confocal microscope. These results indicate that the designed HFI system can be applied to practical biological and medical applications.

  13. Near-infrared fluorescence image quality test methods for standardized performance evaluation

    Science.gov (United States)

    Kanniyappan, Udayakumar; Wang, Bohan; Yang, Charles; Ghassemi, Pejhman; Wang, Quanzeng; Chen, Yu; Pfefer, Joshua

    2017-03-01

    Near-infrared fluorescence (NIRF) imaging has gained much attention as a clinical method for enhancing visualization of cancers, perfusion and biological structures in surgical applications where a fluorescent dye is monitored by an imaging system. In order to address the emerging need for standardization of this innovative technology, it is necessary to develop and validate test methods suitable for objective, quantitative assessment of device performance. Towards this goal, we develop target-based test methods and investigate best practices for key NIRF imaging system performance characteristics including spatial resolution, depth of field and sensitivity. Characterization of fluorescence properties was performed by generating excitation-emission matrix properties of indocyanine green and quantum dots in biological solutions and matrix materials. A turbid, fluorophore-doped target was used, along with a resolution target for assessing image sharpness. Multi-well plates filled with either liquid or solid targets were generated to explore best practices for evaluating detection sensitivity. Overall, our results demonstrate the utility of objective, quantitative, target-based testing approaches as well as the need to consider a wide range of factors in establishing standardized approaches for NIRF imaging system performance.

  14. Homogeneous DNA Detection Based on Fluorescence Quenching by Nanoparticles in Single-step Format :Target-Induced Configuration Transform

    Institute of Scientific and Technical Information of China (English)

    ZHANG,Songbai; WU,Zaisheng; XIE,Ming; SHEN,Guoli; YU,Ruqin

    2009-01-01

    A new strategy for homogeneous detection of DNA hybridization in single-step format was developed based on fluorescence quenching by gold nanoparticles.The gold nanoparticle is functionalized with 5'-thiolated 48-base oligonucleotide(probe sequence),whose 3'-terminus is labeled with fluorescein(FAM),a negatively charged fluo-rescencc dye.The oligonucleotide adopts all extended configuration due to the electrostatic repulsion between nega-tively charged gold nanoparticle and the FAM-attached probe sequence.After addition of the complementary target sequence,specific DNA hybridization induces a conformation change of the probe from an extended stmcture to an arch.1ike configuration,which brings the fluorophore and the gold nanoparticle in close proximity.The fluorescence is efficiently quenched by gold nanoparticles.The fluorescence quenching efficiency is related to the target concen.tration,which allows the quantitative detection for target sequence in a sample.A linear detection range from 1.6 to 209.4 nmoI/L Was obtained under the optimized experimental conditions with a detection limit of 0.1 nmol,L.In the assay system,the gold nanoparticles act as both nanoscaffolds and nanoquenchers.Furthermore,the proposed strategy.in which only two DNA sequences arc involved,is not only different from the traditional molecular bea-cons or reverse molecular beacons but also different from the commonly used sandwich hybridization methods.In addition,the DNA hybridization detection was achieved in homogenous solution in a single-step format,which al-lows real.time detection and quantification with other advantages such as easy operation and elimination of washing steps.

  15. Design and evaluation of capillary coupled with optical fiber light-emitting diode induced fluorescence detection for capillary electrophoresis.

    Science.gov (United States)

    Ji, Hongyun; Li, Meng; Guo, Lihong; Yuan, Hongyan; Wang, Chunling; Xiao, Dan

    2013-09-01

    A new detector, capillary coupled with optical fiber LED-induced fluorescence detector (CCOF-LED-IFD, using CCOF for short), is introduced for CE. The strategy of the present work was that the optical fiber and separation capillary were, in the parallel direction, fastened in a fixation capillary with larger inner diameter. By employing larger inner diameter, the fixation capillary allowed the large diameter of the optical fiber to be inserted into it. By transmitting an enhanced excitation light through the optical fiber, the detection sensitivity was improved. The advantages of the CCOF-CE system were validated by the detection of riboflavin, and the results were compared to those obtained by the in-capillary common optical fiber LED-induced fluorescence detector (IC-COF-LED-IFD, using COF for short). The LODs of CCOF-CE and COF-CE were 0.29 nM and 11.0 nM (S/N = 3), respectively. The intraday (n = 6) repeatability and interday (n = 6) reproducibility of migration time and corresponding peak area for both types of CE were all less than 1.10 and 3.30%, respectively. The accuracy of the proposed method was judged by employing standard addition method, and recoveries obtained were in the range of 98.0-102.4%. The results indicated that the sensitivity of the proposed system was largely improved, and that its reproducibility and accuracy were satisfactory. The proposed system was successfully applied to separate and determine riboflavin in real sample.

  16. Kerosene detection using laser induced fluorescence imaging for aeronautical engines application; Detection du kerozene par imagerie de fluorescence induite par laser, pour application sur foyer aeronautique

    Energy Technology Data Exchange (ETDEWEB)

    Baranger, Ph.

    2004-10-15

    The new concepts of aeronautical engines, developed to follow the evolution of the European standards of pollution, are generally based on an improvement of the processes of liquid fuel injection and mixture in the combustion chamber. There is currently no model mature enough to work without experimental validation. The purpose of this thesis is to assess the possibility of measuring the kerosene (Jet A1) vapour distribution by PLIF (Planar Laser Induced Fluorescence). That measurement technique must quantitatively image the instantaneous concentrations fields of the vaporized fuel in a spray. The implementation of such a technique needs an experimental spectroscopic study, which was realized on the vapour of fuel. First of all, this study allowed us to determine the properties of the kerosene fluorescence spectrum versus physical parameters such as temperature, pressure or gas mixture composition, especially in presence of oxygen molecules. Then, it was shown that the fluorescence spectrum of the fuel could be reproduce in all physical conditions by a single mixture of four aromatics. Their photophysical properties were also analyzed. Following this spectroscopic study, a phenomenological model for the fluorescence of the gaseous fuel was set up. This model led us to a protocol for an optical diagnostic on this fuel vapour. An experiment was set up to test the implementation and the limits of this technique in simple laboratory conditions. This experiment confirmed that this is indeed a promising technique for the diagnostic of the fuel vapour in aeronautical engine. (author)

  17. Optical imaging of breast tumors and of gastrointestinal cancer by laser-induced fluorescence.

    Science.gov (United States)

    Ebert, Bernd; Grosenick, Dirk

    2013-01-01

    Optical imaging offers a high potential for noninvasive detection of cancer in humans. Recent advances in instrumentation for diffuse optical imaging have led to new capabilities for the detection of cancer in highly scattering tissue such as the female breast. We review recent developments in the detection of breast cancer in humans by fluorescent contrast agents. So far, the unspecific contrast agents indocyanine green (ICG) and omocyanine have been applied, whereas molecular probes for direct targeted imaging of this disease are still in preclinical research. We discuss recent improvements in the differentiation of malignant and benign lesions with ICG based on its enhanced extravasation in breast cancer. Whereas fluorescence imaging in thick tissue layers is hampered by strong light scattering, tissue surfaces can be investigated with high spatial resolution. As an example for superficial tumors, lesions of the gastrointestinal tract (GI) are discussed. In these investigations, protoporphyrin IX is used as a tumor-specific (due to its strong enhancement in tumor cells) target for spectroscopic identification and imaging. We present a time-gated method for fluorescence imaging and spectroscopy with strong suppression of tissue autofluorescence and show results on patients with Barrett's esophagus and with colitis ulcerosa.

  18. Highly sensitive analysis of flavonoids by zwitterionic microemulsion electrokinetic chromatography coupled with light-emitting diode-induced fluorescence detection.

    Science.gov (United States)

    Cao, Wan; Hu, Shuai-Shuai; Li, Xing-Ying; Pang, Xiao-Qing; Cao, Jun; Ye, Li-Hong; Dai, Han-Bin; Liu, Xiao-Juan; Da, Jian-Hua; Chu, Chu

    2014-09-05

    A rapid zwitterionic microemulsion electrokinetic chromatography (ZI-MEEKC) approach coupled with light-emitting-diode-induced fluorescence (LED-IF, 480nm) detection was proposed for the analysis of flavonoids. In the optimization process, we systematically investigated the separation conditions, including the surfactants, cosurfactants, pH, buffers and fluorescence parameters. It was found that the baseline separation of the seven flavonoids was obtained in less than 5min with a running buffer consisting of 92.9% (v/v) 5mM sodium borate, 0.6% (w/v) ZI surfactant, 0.5% (w/v) ethyl acetate and 6.0% (w/v) 1-butanol. High sensitivity was obtained by the application of LED-IF detection. The limits of detection for seven flavonoids were in the range of 3.30×10(-8) to 2.15×10(-6)molL(-1) without derivatization. Ultimately, the detection method was successfully applied to the analysis of flavonoids in hawthorn plant and food products with satisfactory results. Copyright © 2014 Elsevier B.V. All rights reserved.

  19. Förster resonance energy transfer and protein-induced fluorescence enhancement as synergetic multi-scale molecular rulers

    Science.gov (United States)

    Ploetz, Evelyn; Lerner, Eitan; Husada, Florence; Roelfs, Martin; Chung, Sangyoon; Hohlbein, Johannes; Weiss, Shimon; Cordes, Thorben

    2016-09-01

    Advanced microscopy methods allow obtaining information on (dynamic) conformational changes in biomolecules via measuring a single molecular distance in the structure. It is, however, extremely challenging to capture the full depth of a three-dimensional biochemical state, binding-related structural changes or conformational cross-talk in multi-protein complexes using one-dimensional assays. In this paper we address this fundamental problem by extending the standard molecular ruler based on Förster resonance energy transfer (FRET) into a two-dimensional assay via its combination with protein-induced fluorescence enhancement (PIFE). We show that donor brightness (via PIFE) and energy transfer efficiency (via FRET) can simultaneously report on e.g., the conformational state of double stranded DNA (dsDNA) following its interaction with unlabelled proteins (BamHI, EcoRV, and T7 DNA polymerase gp5/trx). The PIFE-FRET assay uses established labelling protocols and single molecule fluorescence detection schemes (alternating-laser excitation, ALEX). Besides quantitative studies of PIFE and FRET ruler characteristics, we outline possible applications of ALEX-based PIFE-FRET for single-molecule studies with diffusing and immobilized molecules. Finally, we study transcription initiation and scrunching of E. coli RNA-polymerase with PIFE-FRET and provide direct evidence for the physical presence and vicinity of the polymerase that causes structural changes and scrunching of the transcriptional DNA bubble.

  20. Förster resonance energy transfer and protein-induced fluorescence enhancement as synergetic multi-scale molecular rulers

    Science.gov (United States)

    Ploetz, Evelyn; Lerner, Eitan; Husada, Florence; Roelfs, Martin; Chung, SangYoon; Hohlbein, Johannes; Weiss, Shimon; Cordes, Thorben

    2016-01-01

    Advanced microscopy methods allow obtaining information on (dynamic) conformational changes in biomolecules via measuring a single molecular distance in the structure. It is, however, extremely challenging to capture the full depth of a three-dimensional biochemical state, binding-related structural changes or conformational cross-talk in multi-protein complexes using one-dimensional assays. In this paper we address this fundamental problem by extending the standard molecular ruler based on Förster resonance energy transfer (FRET) into a two-dimensional assay via its combination with protein-induced fluorescence enhancement (PIFE). We show that donor brightness (via PIFE) and energy transfer efficiency (via FRET) can simultaneously report on e.g., the conformational state of double stranded DNA (dsDNA) following its interaction with unlabelled proteins (BamHI, EcoRV, and T7 DNA polymerase gp5/trx). The PIFE-FRET assay uses established labelling protocols and single molecule fluorescence detection schemes (alternating-laser excitation, ALEX). Besides quantitative studies of PIFE and FRET ruler characteristics, we outline possible applications of ALEX-based PIFE-FRET for single-molecule studies with diffusing and immobilized molecules. Finally, we study transcription initiation and scrunching of E. coli RNA-polymerase with PIFE-FRET and provide direct evidence for the physical presence and vicinity of the polymerase that causes structural changes and scrunching of the transcriptional DNA bubble. PMID:27641327

  1. Human neutrophil elastase inhibition studied by capillary electrophoresis with laser induced fluorescence detection and microscale thermophoresis.

    Science.gov (United States)

    Syntia, Fayad; Nehmé, Reine; Claude, Bérengère; Morin, Philippe

    2016-01-29

    Capillary electrophoresis-laser induced fluorescence (CZE-LIF) and microscale thermophoresis (MST) were used for the first time to study the inhibition of human neutrophil elastase (HNE). We recently studied HNE kinetics (Km and Vmax) by developing an in-capillary CZE-LIF assay based on transverse diffusion of laminar flow profiles (TDLFP) for reactant mixing. In this work, the former assay was adapted to monitor HNE inhibition. Two natural well known HNE inhibitors from the triterpene family, ursolic acid and oleanolic acid, were tested to validate the developed assay. Since the solubility of pentacyclic triterpenes in aqueous media where the enzymatic reaction will take place is limited, the effect of DMSO and ethanol on HNE was studied using microscale thermophoresis (MST). An agglomeration of the enzyme was revealed when preparing the inhibitor in 5% (v/v) DMSO. This phenomenon did not occur in the presence of ethanol. Therefore, ethanol was used as inhibitor solvent, at a limited percentage of 20% (v/v). In these conditions and after optimization of the TDLFP approach, the repeatability (RSD on migration times and peak-areas inferior to 2.2%) of the CZE-LIF assay and the sensitivity (LOQ of few nM) were found to be satisfactory for conducting inhibition assays. IC50 values for ursolic and oleanolic acid were successfully determined. They were respectively equal to 5.62±0.10μM (r(2)=0.9807; n=3) and to 8.21±0.23μM (r(2)=0.9887; n=3). Excellent agreement was found between the results obtained by CE and those reported in literature which validates the developed method. Particularly, the CE-based assay is able to rank HNE inhibitors relative to each other. Furthermore, MST technique was used for evaluating HNE interaction with the ursolic acid. Up to 16 capillaries were automatically processed to obtain in one titration experiment the dissociation constant for the HNE-ursolic acid complex. Ki was found to be 2.72±0.66μM (n=3) which is in excellent agreement

  2. Fast methods for analysis of neurotransmitters from single cell and monitoring their releases in central nervous system by capillary electrophoresis, fluorescence microscopy and luminescence imaging

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ziqiang [Iowa State Univ., Ames, IA (United States)

    1999-12-10

    Fast methods for separation and detection of important neurotransmitters and the releases in central nervous system (CNS) were developed. Enzyme based immunoassay combined with capillary electrophoresis was used to analyze the contents of amino acid neurotransmitters from single neuron cells. The release of amino acid neurotransmitters from neuron cultures was monitored by laser induced fluorescence imaging method. The release and signal transduction of adenosine triphosphate (ATP) in CNS was studied with sensitive luminescence imaging method. A new dual-enzyme on-column reaction method combined with capillary electrophoresis has been developed for determining the glutamate content in single cells. Detection was based on monitoring the laser-induced fluorescence of the reaction product NADH, and the measured fluorescence intensity was related to the concentration of glutamate in each cell. The detection limit of glutamate is down to 10-8 M level, which is 1 order of magnitude lower than the previously reported detection limit based on similar detection methods. The mass detection limit of a few attomoles is far superior to that of any other reports. Selectivity for glutamate is excellent over most of amino acids. The glutamate content in single human erythrocyte and baby rat brain neurons were determined with this method and results agreed well with literature values.

  3. Fast methods for analysis of neurotransmitters from single cell and monitoring their releases in central nervous system by capillary electrophoresis, fluorescence microscopy and luminescence imaging

    Energy Technology Data Exchange (ETDEWEB)

    Wang, Ziqiang

    1999-12-10

    Fast methods for separation and detection of important neurotransmitters and the releases in central nervous system (CNS) were developed. Enzyme based immunoassay combined with capillary electrophoresis was used to analyze the contents of amino acid neurotransmitters from single neuron cells. The release of amino acid neurotransmitters from neuron cultures was monitored by laser induced fluorescence imaging method. The release and signal transduction of adenosine triphosphate (ATP) in CNS was studied with sensitive luminescence imaging method. A new dual-enzyme on-column reaction method combined with capillary electrophoresis has been developed for determining the glutamate content in single cells. Detection was based on monitoring the laser-induced fluorescence of the reaction product NADH, and the measured fluorescence intensity was related to the concentration of glutamate in each cell. The detection limit of glutamate is down to 10{sup {minus}8} M level, which is 1 order of magnitude lower than the previously reported detection limit based on similar detection methods. The mass detection limit of a few attomoles is far superior to that of any other reports. Selectivity for glutamate is excellent over most of amino acids. The glutamate content in single human erythrocyte and baby rat brain neurons were determined with this method and results agreed well with literature values.

  4. Extraction and optical fluorescence method for the measurement of trace beryllium in soils.

    Science.gov (United States)

    Agrawal, Anoop; Cronin, John P; Agrawal, Akshay; Tonazzi, Juan C L; Adams, Lori; Ashley, Kevin; Brisson, Michael J; Duran, Brandy; Whitney, Gary; Burrell, Anthony K; McCleskey, T Mark; Robbins, James; White, Kenneth T

    2008-03-15

    Beryllium metal and beryllium oxide are important industrial materials used in a variety of applications in the electronics, nuclear energy, and aerospace industries. These materials are highly toxic, they must be disposed of with care, and exposed workers need to be protected. Recently, a new analytical method was developed that uses dilute ammonium bifluoride for extraction of beryllium and a high quantum yield optical fluorescence reagent to determine trace amounts of beryllium in airborne and surface samples. The sample preparation and analysis procedure was published by both ASTM International and the National Institute for Occupational Safety and Health (NIOSH). The main advantages of this method are its sensitivity, simplicity, use of lower toxicity materials, and low capital costs. Use of the technique for analyzing soils has been initiated to help meet a need at several of the U.S. Department of Energy legacy sites. So far this work has mainly concentrated on developing a dissolution protocol for effectively extracting beryllium from a variety of soils and sediments so that these can be analyzed by optical fluorescence. Certified reference materials (CRM) of crushed rock and soils were analyzed for beryllium content using fluorescence, and results agree quantitatively with reference values.

  5. [Commercial orange juice beverages detection by fluorescence spectroscopy combined with PCA-ED and PLSR methods].

    Science.gov (United States)

    Hu, Yang-jun; Zhu, Chun; Chen, Guo-qing; Zhang, Yong; Kong, Fan-biao; Li, Run; Zhu, Zhuo-wei; Wang, Xu; Gao, Shu-mei

    2014-08-01

    In order to classify the orange juiice beverages effectively, the fluorescence character differences of two kinds of orange juice beverages including 100% orange juice and orange drink were analyzed and compared, principal component analysis combined with Euclidean distance was adopted to classify two kinds of orange juice beverages, and ideal classification results were obtained. Meanwhile, the orange juice content estimation model was established by using fluorescence spectroscopy combined with partial least squares regression method, and the correlation coefficient R, root mean square error of calibration RMSEC and root mean square error of prediction RMSEP were 0.997, 0.87% and 2.05%, respectively. The experimental results indicate that the calibration model offers comparatively accurate content estimation, which reflect the actual orange juice content in the commercial orange juice beverages. The exploration to classify orange juice beverages was carried out from two aspects of qualitative and quantitative analysis by employing fluorescence spectroscopy combined with chemometrics method, which can provide a new idea for the classification and adulteration detection of commercial orange juice beverages, and also can give certain reference basis for the quality control of orange juice raw material.

  6. Study of Sugar Cane Management Systems in Brazil Using Laser Induced Fluorescence

    Science.gov (United States)

    Cabral, Jader; Villas-Boas, Paulino; Carvalho, Camila; Corá, José Eduardo; Milori, Débora

    2014-05-01

    Brazil is the largest producer of cane sugar, consequently, is a leader in the production of bio-ethanol, a clean and renewable energy that fits the model of sustainable economy as discussed and pursued by our society. Our state of São Paulo concentrates 60% of national production, representing a sizeable share in the range of world production. All this economic potential is closely monitored by the scientific community, which develops numerous studies seeking an improvement in production efficiency and reduced environmental impacts caused by the planting. However, the study of soil samples, in plantation areas, demands results about the content and structural forms of organic matter (OM). Also, the soil carbon stocks depend on the type of management. Our goal is to study OM of soil samples from four sugar cane management systems: (i) unburned cane harvest, (ii) preharvest burned, (iii) addition of sugarcane bagasse ash and (iv) addition of residue from the extraction of sucrose, using Laser Induced Fluorescence Spectroscopy of solid state. All the emission spectra were acquired using the system called LIFS-405, which consists of a diode laser Coherent, model cube with excitation at 405 nm, maximum output power of 50mJ and a mini-spectrometer, Ocean Optics USB2000-high sensitivity, with range of 194-894 nm and a fiber-optic bundle design (six excitation fibers in a circular path and one central fiber the collect the fluorescence). In this work, we will present the preliminary results evolving the humification index (HLIFS) of soil OM and total carbon amount (TC) for the different types of management. HLIFS shows a close correlation with the humification index of humic acid in solution obtained by means 2D conventional fluorescence spectroscopy.

  7. Stepwise multi-photon activation fluorescence reveals a new method of melanoma imaging for dermatologists

    Science.gov (United States)

    Lai, Zhenhua; Lian, Christine; Ma, Jie; Yu, Jingyi; Gu, Zetong; Rajadhyaksha, Milind; DiMarzio, Charles A.

    2014-02-01

    Previous research has shown that the stepwise multi-photon activated fluorescence (SMPAF) of melanin, activated by a continuous-wave (CW) mode near infrared (NIR) laser, is a low cost and reliable method of detecting melanin. SMPAF images of melanin in a mouse hair and a formalin fixed mouse melanoma were compared with conventional multiphoton fluorescence microscopy (MPFM) images and confocal reflectance microscopy (CRM) images, all of which were acquired at an excitation wavelength of 920 nm, to further prove the effectiveness of SMPAF in detecting melanin. SMPAF images add specificity for melanin detection to MPFM images and CRM images. Melanin SMPAF can be a promising technology to enable melanoma imaging for dermatologists.

  8. Interphase cytogenetics in pathology: principles, methods, and applications of fluorescence in situ hybridization (FISH).

    Science.gov (United States)

    Werner, M; Wilkens, L; Aubele, M; Nolte, M; Zitzelsberger, H; Komminoth, P

    1997-01-01

    Characteristic chromosome aberrations have been identified in various tumors. Fluorescence in situ hybridization (FISH) using specific probes that are generated by vector cloning or in vitro amplification and labeled with fluorescent dyes allow for the detection of these genetic changes in interphase cells. This technique, that is also referred to as "interphase cytogenetics", can be performed in cytological preparations as well as in sections of routinely formaldehyde-fixed and paraffin-embedded tissue. In cancer research and diagnostics, interphase cytogenetics by FISH is used to detect numerical chromosome changes and structural aberrations, e.g., translocations, deletions, or amplifications. In this technical overview, we explain the principles of the FISH method and provide protocols for FISH in cytological preparations and paraffin sections. Moreover, possible applications of FISH are discussed.

  9. Fluorescence fluctuation microscopy: a diversified arsenal of methods to investigate molecular dynamics inside cells.

    Science.gov (United States)

    Weidemann, Thomas; Mücksch, Jonas; Schwille, Petra

    2014-10-01

    Fluorescence microscopy provides insight into the subcellular organization of biological functions. However, images are snap shots averaging over a highly dynamic molecular system. Fluorescence fluctuation microscopy, employing similar detection technology, encompasses a powerful arsenal of analysis tools that investigate the molecular heterogeneity in space and time. Analyzing signal fluctuations from small ensembles (several hundred particles) reveals their concentration, the stoichiometry, the stochastic motion, as well as superimposed signatures of the environment such as spatial confinement and binding events. Thus, fluctuation analysis provides access to dynamic molecular properties that can be used to build physical models of cellular processes. In the last decade these methods experienced a remarkable diversification, which we revisit here with a particular focus on live cell applications.

  10. A solvothermal method to synthesize fluorescent carbon nanoparticles and application to photocatalysis and electrocatalysis.

    Science.gov (United States)

    Hongren, Li; Feng, Li; Aimin, Deng

    2015-09-01

    A novel solvothermal approach to synthesize fluorescent carbon nanoparticles (CNPs) was developed using glucose and ammonium oxalate as the carbon source, and glycol as the solvent. The solution of as-prepared CNPs emitted blue-green fluorescence under ultraviolet (UV) light at 365 nm, and the carbon nanoparticle formation was investigated through XRD, TEM, DLS, FT-IR, UV, PL, XPS. The particle was well dispersed with an average diameter of about 10-30 nm. In contrast with previous methods, in this synthesis process neither strong acid treatment nor further surface modification was necessary. The solution of as-prepared CNPs were applied to photocatalytic degradation of mountain green in the present H2 O2 , and the decolorization rate was over 97% when the reaction time was more than 7 h under visible light. The as-prepared CNPs were also applied to electrocatalysis and showed excellent electrocatalytic activity.

  11. A new screening method for flunitrazepam in vodka and tequila by fluorescence spectroscopy.

    Science.gov (United States)

    Leesakul, Nararak; Pongampai, Sirintip; Kanatharana, Proespichaya; Sudkeaw, Pravit; Tantirungrotechai, Yuthana; Buranachai, Chittanon

    2013-01-01

    A new screening method for flunitrazepam in colourless alcoholic beverages based on a spectroscopic technique is proposed. Absorption and steady-state fluorescence of flunitrazepam and its protonated form with various acids were investigated. The redshift of the wavelength of maximum absorption was distinctively observed in protonated flunitrazepam. An emissive fluorescence at 472 nm was detected in colourless spirits (vodka and tequila) at room temperature. 2-M perchloric acid was the most appropriated proton source. By using electron ionization mass spectrometry and time-dependent density functional theory calculations, the possible structure of protonated flunitrazepam was identified to be 2-nitro-N-methylacridone, an acridone derivative as opposed to 2-methylamino-5-nitro-2'-fluorobenzophenone, a benzophenone derivative.

  12. Establishing a cellular FRET-based fluorescence plate reader assay to monitor proNGF-induced cross-linking of sortilin and the neurotrophin receptor p75(NTR).

    Science.gov (United States)

    Skeldal, Sune; Kjaergaard, Maj M; Alwasel, Saleh; Nyengaard, Jens R

    2015-01-01

    Whereas the proform of the nerve growth factor (proNGF) is crucial for eliminating superfluous cells during neuronal development it also promotes apoptosis following brain trauma and neuronal injury. The apoptotic signal is elicited upon formation of a trimeric receptor complex also containing the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay to monitor the interaction between fluorescently tagged sortilin and p75(NTR) in live cells. The method is based on a standard fluorescent plate reader found in many biochemical laboratories and the results are evaluated using a microscopy-based quantified sensitized acceptor emission FRET approach making use of a pair of FRET standard constructs. As a result, the effect of proNGF on the interaction between sortilin and p75(NTR) can be evaluated in live cells allowing for screening and selection of therapeutic compounds interfering with proNGF-induced cell death.

  13. Design and development of a LabVIEW-based LED-induced fluorescence spectroscopy system with applications in non-destructive quality assessment of agricultural products

    Science.gov (United States)

    Abbasi, Hamed; Nazeri, Majid; Mireei, Seyed Ahmad

    2016-01-01

    Over the past several years, the demand for high quality agricultural products has been remarkably increased. Thus, it is important to use non-destructive methods for product quality monitoring. LED-induced fluorescence spectroscopy has proved its potential for nondestructive detection of some defects in agricultural products, such as tissue browning and bruising. Due to such defects, changes in the polyphenol and chlorophyll contents occur which can be considered as the visible marks of decreasing fruit quality. In the present work, a fluorescence spectrometer (spectrofluorometer) controlled by LabVIEW software was designed and developed. In this spectrometer, a consumer-grade webcam was used as an imaging sensor. The spectrometer was able to measure the fluorescence spectra directly from the fruit and vegetable surface in the desired regions. To do so, the spectrometer was equipped with a suitable fiber-optic probe. The hardware solution was based on data acquisition working on the USB platform and controlled by the application running on the PC. In this system, light emitting diodes with different wavelengths were used as the excitation sources for inducing fluorescence spectra of some famous fruits and vegetables.

  14. Fluorescence Resonance Energy Transfer Analysis of Bid Activation in Living Cells during Ultraviolet-induced Apoptosis

    Institute of Scientific and Technical Information of China (English)

    Yinyuan WU; Da XING; Lei LIU; Tongsheng CHEN; Wei R. CHEN

    2007-01-01

    Ultraviolet (UV) irradiation is a DNA-damaging agent that triggers apoptosis through both the membrane death receptor and mitochondrial apoptotic signaling pathways. Bid, a pro-apoptotic Bcl-2family member, is important in most cell types to apoptosis in response to DNA damage. In this study, a recombinant plasmid, YFP-Bid-CFP, comprised of yellow and cyan fluorescent protein and a full length Bid,was used as a fluorescence resonance energy transfer analysis (FRET) probe. Using the FRET technique based on YFP-Bid-CFP, we found that Bid activation was initiated at 9±1 h after UV irradiation, and the average duration of the activation was 75± 10 min. Bid activation coincided with a collapse of the mitochondrial membrane potential with an average duration of 50±10 min. When cells were pretreated with Z-IETD-fmk(caspase-8 specific inhibitor) the process of Bid activation was completely inhibited, but the apoptosis was only partially affected. Z-DEVD-fmk (caspase-3 inhibitor) and Z-FA-fmk (non asp specific inhibitor) did not block Bid activation. Furthermore, the endogenous Bid activation with or without Z-IETD-fmk in response to UV irradiation was confirmed by Western blotting. In summary, using the FRET technique, we observed the dynamics of Bid activation during UV-induced apoptosis and found that it was a caspase-8 dependent event.

  15. Comparison of Sun-Induced Chlorophyll Fluorescence Estimates Obtained from Four Portable Field Spectroradiometers

    Directory of Open Access Journals (Sweden)

    Tommaso Julitta

    2016-02-01

    Full Text Available Remote Sensing of Sun-Induced Chlorophyll Fluorescence (SIF is a research field of growing interest because it offers the potential to quantify actual photosynthesis and to monitor plant status. New satellite missions from the European Space Agency, such as the Earth Explorer 8 FLuorescence EXplorer (FLEX mission—scheduled to launch in 2022 and aiming at SIF mapping—and from the National Aeronautics and Space Administration (NASA such as the Orbiting Carbon Observatory-2 (OCO-2 sampling mission launched in July 2014, provide the capability to estimate SIF from space. The detection of the SIF signal from airborne and satellite platform is difficult and reliable ground level data are needed for calibration/validation. Several commercially available spectroradiometers are currently used to retrieve SIF in the field. This study presents a comparison exercise for evaluating the capability of four spectroradiometers to retrieve SIF. The results show that an accurate far-red SIF estimation can be achieved using spectroradiometers with an ultrafine resolution (less than 1 nm, while the red SIF estimation requires even higher spectral resolution (less than 0.5 nm. Moreover, it is shown that the Signal to Noise Ratio (SNR plays a significant role in the precision of the far-red SIF measurements.

  16. Laser Induced Dual Fluorescence Ratiometric Technique for Mixing Characterization in Microfluidic Systems

    Science.gov (United States)

    Bedding, David; Hidrovo, Carlso

    2016-11-01

    Increasing the rate of mixing within microfluidic systems is vitally important in understanding biological and chemical reaction kinetics and mechanisms. The small length scales characteristic of these systems which translate into highly viscous, Stokes flows result in mixing that is primarily dominated by diffusion. In order to counteract this, an approach that utilizes inertial droplet collisions to promote chaotic advection between two mixing species has been developed. A Laser-Induced Dual Fluorescence (LIDF) system in conjunction with a high-speed camera and appropriate optics are used to capture two intensity fields providing information about the mixing process as well as the excitation intensity field over the volume of interest. The rate of mixing for the coalescing droplets was quantified by taking the standard deviation of the first intensity field over time, while the second intensity field provides information about the intensity field. A ratiometric imaging approach allows removal of mixing fluorescence signal noise in the form of variation in excitation intensity, primarily from the lasing patterns and lensing effects within the interrogation volume. NSF CAREER Award Grant CBET - 1151091.

  17. Light induced fluorescence for predicting API content in tablets: sampling and error.

    Science.gov (United States)

    Domike, Reuben; Ngai, Samuel; Cooney, Charles L

    2010-05-31

    The use of a light induced fluorescence (LIF) instrument to estimate the total content of fluorescent active pharmaceutical ingredient in a tablet from surface sampling was demonstrated. Different LIF sampling strategies were compared to a total tablet ultraviolet (UV) absorbance test for each tablet. Testing was completed on tablets with triamterene as the active ingredient and on tablets with caffeine as the active ingredient, each with a range of concentrations. The LIF instrument accurately estimated the active ingredient within 10% of total tablet test greater than 95% of the time. The largest error amongst all of the tablets tested was 13%. The RMSEP between the techniques was in the range of 4.4-7.9%. Theory of the error associated with the surface sampling was developed and found to accurately predict the experimental error. This theory uses one empirically determined parameter: the deviation of estimations at different locations on the tablet surface. As this empirical parameter can be found rapidly, correct use of this prediction of error may reduce the effort required for calibration and validation studies of non-destructive surface measurement techniques, and thereby rapidly determine appropriate analytical techniques for estimating content uniformity in tablets.

  18. Detection of adenosine triphosphate in HeLa cell using capillary electrophoresis-laser induced fluorescence detection based on aptamer and graphene oxide.

    Science.gov (United States)

    Fang, Bi-Yun; Yao, Ming-Hao; Wang, Chun-Yuan; Wang, Chao-Yang; Zhao, Yuan-Di; Chen, Fang

    2016-04-01

    A method for ATP quantification based on dye-labeled aptamer/graphene oxide (aptamer/GO) using capillary electrophoresis-laser induced fluorescence (CE-LIF) detecting technique has been established. In this method, the carboxyfluorescein (FAM)-labelled ATP aptamers were adsorbed onto the surface of GO, leading to the fluorescence quenching of FAM; after the incubation with a limited amount of ATP, stronger affinity between ATP aptamer and ATP resulted in the desorption of aptamers and the fluorescence restoration of FAM. Then, aptamer-ATP complex and excess of aptamer/GO and GO were separated and quantified by CE-LIF detection. It was shown that a linear relation was existing in the CE-LIF peak intensity of aptamer-ATP and ATP concentration in range of 10-700 μM, the regression equation was F=1.50+0.0470C(ATP) (R(2)=0.990), and the limit of detection was 1.28 μM (3S/N, n=5), which was one order magnitude lower than that of detection in solution by fluorescence method. The approach with excellent specificity and reproducibility has been successfully applied to detecting concentration of ATP in HeLa cell.

  19. Imaging and imagining the spatio-temporal variations of photosynthesis - remote sensing of sun-induced fluorescence to understand physiological changes of the photosynthetic apparatus

    Science.gov (United States)

    Rascher, Uwe

    2010-05-01

    Light use efficiency of photosynthesis dynamically adapts to environmental factors, which lead to complex spatio-temporal variations of photosynthesis on various scales from the leaf to the canopy level. The need to scale leaf-level physiology to ecosystem responses and climate feedbacks has been emphasized recently in the context of global climate change research. Recently the FLuorescence EXplorer (FLEX) mission that proposed to launch a satellite for the global monitoring of steady-state chlorophyll fluorescence in terrestrial vegetation was selected for pre-phase A by European Space Agency (ESA). This method aims for mapping photosynthetic efficiency by quantifying steady state fluorescence in the so called Fraunhofer lines. In preparation for this satellite mission an extensive field campaign was conducted. The CEFLES2 campaign during the Carbo Europe Regional Experiment Strategy was designed to provide simultaneous airborne measurements of solar induced fluorescence and CO2 fluxes. It was combined with extensive ground-based quantification of leaf- and canopy-level processes. The aim of this campaign was to test if fluorescence signal detected from an airborne platform can be used to improve estimates of plant mediated exchange on the mesoscale. Canopy fluorescence was quantified from four airborne platforms using a combination of novel sensors including a prototype airborne sensor AirFLEX quantifing fluorescence in the oxygen A and B bands, and the first employment of the high performance imaging spectrometer HYPER delivering spatially explicit and multi-temporal transects across the whole region. During three measurement periods in April, June and September 2007 structural, functional and radiometric characteristics of more than 20 different vegetation types in the Les Landes region, Southwest France, were extensively characterized on the ground focussing especially on quantifying plant mediated exchange processes (photosynthetic electron transport, CO2

  20. Protective effect of light emitting diode phototherapy on fluorescent light induced retinal damage in Wistar strain albino rats.

    Science.gov (United States)

    Ahamed Basha, A; Mathangi, D C; Shyamala, R; Ramesh Rao, K

    2014-09-01

    Artificial light at night alters retinal physiology. Several studies have shown that light emitting diode phototherapy protects the retina from the damaging effects of acute light exposure. The aim of this study has been to elucidate the protective effects of 670 nm LED light on retinal damage induced by chronic fluorescent light in Wistar rats. Male Wistar albino rats were divided into four groups: group 1 were control (CL), group 2, 3 and 4 were exposed to fluorescent light (FL), LED preexposure+fluorescent light exposure (LL) and only LED light exposure (OL) respectively. All animals were maintained in their specific exposure regime for 30 days. Fluorescent light of 1800 lx was exposed between 8 pm to 8 am. Rats were exposed to therapeutic LED light of 670 nm of 9 J/cm2 at 25 mW/cm2 for 6 min duration. Histopathological changes in the retina were studied. Animals of the FL group showed a significant reduction in the outer nuclear layer thickness and cell count in addition to the total thickness of the retina. LL group which were exposed to 670 nm LED prior to exposure to fluorescent light showed a significant decrease in the degree of damage. 670 nm LED light preexposure is protective to retinal cells against fluorescent light-induced damage. Copyright © 2014 Elsevier GmbH. All rights reserved.

  1. Effect of LED photobiomodulation on fluorescent light induced changes in cellular ATPases and Cytochrome c oxidase activity in Wistar rat.

    Science.gov (United States)

    A, Ahamed Basha; C, Mathangi D; R, Shyamala

    2016-12-01

    Fluorescent light exposure at night alters cellular enzyme activities resulting in health defects. Studies have demonstrated that light emitting diode photobiomodulation enhances cellular enzyme activities. The objectives of this study are to evaluate the effects of fluorescent light induced changes in cellular enzymes and to assess the protective role of pre exposure to 670 nm LED in rat model. Male Wistar albino rats were divided into 10 groups of 6 animals each based on duration of exposure (1, 15, and 30 days) and exposure regimen (cage control, exposure to fluorescent light [1800 lx], LED preexposure followed by fluorescent light exposure and only LED exposure). Na(+)-K(+) ATPase, Ca(2+) ATPase, and cytochrome c oxidase of the brain, heart, kidney, liver, and skeletal muscle were assayed. Animals of the fluorescent light exposure group showed a significant reduction in Na(+)-K(+) ATPase and Ca(2+) ATPase activities in 1 and 15 days and their increase in animals of 30-day group in most of the regions studied. Cytochrome c oxidase showed increase in their level at all the time points assessed in most of the tissues. LED light preexposure showed a significant enhancement in the degree of increase in the enzyme activities in almost all the tissues and at all the time points assessed. This study demonstrates the protective effect of 670 nm LED pre exposure on cellular enzymes against fluorescent light induced change.

  2. Manipulating quantum dot fluorescence by utilizing Brownian induced near-field interactions with plasmonic nanoparticles

    Science.gov (United States)

    Palombo, Nola

    Quantum dots (QDs) are semiconductor nanocrystals with size-dependent optical properties; thus making them supreme fluorophores. Plasmonic nanoparticles (PNPs), such as gold and silver nanoparticles, support localized surface plasmons on their surface. When the localized surface plasmons are excited, a highly concentrated electromagnetic field is formed near the particle. Therefore, if a QD is within the near-field of a PNP, the emission or excitation of the QD can be enhanced. However, due to Forster Resonance Energy Transfer (FRET), the QD fluorescence could instead be quenched by the proximity of PNPs. Whether enhancement or quenching occurs, is dependent upon the distance and geometry of the nanoparticles. Enhanced QD fluorescence would be helpful in biomedical sensing and imaging and solar energy conversion applications. In addition, quenched QD fluorescence caused by FRET could be applied to FRET-based sensing and imaging in medical diagnosis. This master's thesis first theoretically models the stochastic movement of QDs and PNPs in an aqueous solution. The simulation is based upon the Direct Simulation Monte Carlo method coupled with Langevin equations. Using this simulation, we were able to predict the percentage of QDs in the near-field region of PNPs. The percentage of QDs in the near-field region of GNPs for a concentration of 1 × 1013 QDs/mL and 5 × 108 GNPs/mL, is a very small percentage of 2 × 10-5%. Yet, the concentration of QDs in the near-field region of GNPs was calculated to be 1,510,000 QDs mL-1. In addition, this master's thesis experimentally explores the enhancement and quenching of QD emission for different concentrations and sizes of PNPs in aqueous solutions. The fluorescence spectra of two types of QD-PNP mixtures were measured. The first mixture was QDs and gold nanoparticles (GNPs) dispersed in distilled water, where the emission wavelength of the QDs matches the localized surface plasmon excitation wavelength of the GNPs. The second

  3. Oxygen plasma flow properties deduced from laser-induced fluorescence and probe measurements

    Science.gov (United States)

    Löhle, Stefan; Eichhorn, Christoph; Steinbeck, Andreas; Lein, Sebastian; Herdrich, Georg; Röser, Hans-Peter; Auweter-Kurtz, Monika

    2008-04-01

    Estimation of the local dissociation degree and the local mass-specific enthalpy of a pure oxygen plasma flow determined mainly from laser-induced fluorescence measurements are reported. Measurements have been conducted for several generator parameters in an inductively heated plasma wind tunnel. Additional probe measurements of total pressure together with the deduced translational temperature are used to estimate the local mass-specific enthalpy. For a reference condition, full dissociation has been measured. The measured translational temperature of atomic oxygen for this condition is T = 3500 K. Subsequently, the local mass-specific enthalpy has been derived using these local density and temperature measurements. For the reference condition the estimated value of h = 27 MJ/kg is in good agreement with the probe measurements and results from diode laser absorption spectroscopy.

  4. High resolution isotope shifts and hyperfine structure measurements of tungsten by laser induced fluorescence spectroscopy

    CERN Document Server

    Lee, Jeongwon; Leanhardt, Aaron

    2012-01-01

    Isotope shifts and hyperfine structure of tungsten were studied in the near UV range. We have used laser induced fluorescence spectroscopy on a pulsed supersonic beam to probe the 5D0 -> 5F1 transition at 384.9 nm, 7S3 -> 7P4 transition at 400.9 nm, and 7S3 -> 7P3 transition at 407.4 nm. Three new magnetic hyperfine constants are reported for 7P3,7P4, and 5F1 states. The isotope shifts of the 384.9 nm transition are presented for the first time, and the isotope shifts of 400.9 nm and 407.4 nm transition are measured with an order of magnitude higher precision compared to the previous measurements. As a result, the nuclear parameters lambda and lambda_{rel} are extracted from the isotope shifts with an improved precision.

  5. Tundra photosynthesis captured by satellite-observed solar-induced chlorophyll fluorescence

    Science.gov (United States)

    Luus, K. A.; Commane, R.; Parazoo, N. C.; Benmergui, J.; Euskirchen, E. S.; Frankenberg, C.; Joiner, J.; Lindaas, J.; Miller, C. E.; Oechel, W. C.; Zona, D.; Wofsy, S.; Lin, J. C.

    2017-02-01

    Accurately quantifying the timing and magnitude of respiration and photosynthesis by high-latitude ecosystems is important for understanding how a warming climate influences global carbon cycling. Data-driven estimates of photosynthesis across Arctic regions often rely on satellite-derived enhanced vegetation index (EVI); we find that satellite observations of solar-induced chlorophyll fluorescence (SIF) provide a more direct proxy for photosynthesis. We model Alaskan tundra CO2 cycling (2012-2014) according to temperature and shortwave radiation and alternately input EVI or SIF to prescribe the annual seasonal cycle of photosynthesis. We find that EVI-based seasonality indicates spring "green-up" to occur 9 days prior to SIF-based estimates, and that SIF-based estimates agree with aircraft and tower measurements of CO2. Adopting SIF, instead of EVI, for modeling the seasonal cycle of tundra photosynthesis can result in more accurate estimates of growing season duration and net carbon uptake by arctic vegetation.

  6. A Lagrangian Interpretation of Laser Induced Fluorescence Signals in a Plasma

    Science.gov (United States)

    Chu, Feng; Skiff, Fred; Berumen, Jorge; Mattingly, Sean; Hood, Ryan

    2015-11-01

    Laser induced fluorescence (LIF) is a nonintrusive diagnostic technique that has found applications in the study of a wide range of fundamental and applied problems. Thus it is important to make a correct interpretation of LIF signals. We adopt a Lagrangian approach to model LIF signals by introducing a non-linear conditional probability function P(x,v,tx',v',t'). A simulation is performed to compute the LIF signals and the results are presented. We investigate how mean-field waves affect these signals and metastable state birth rates. The ultimate goal is to construct the complete model for LIF signals by combining optical pumping, mean-field wave effect and metastable state birth rate modulation. This research is supported by the Department of Energy under grant No. DOE DE-FG02-99ER54543.

  7. Average OH density in alternating current dielectric barrier discharge by laser-induced fluorescence technique

    Science.gov (United States)

    Yang, Hongliang; Feng, Chunlei; Gao, Liang; Ding, Hongbin

    2015-10-01

    The average OH density in atmospheric He-H2O(0.4%) needle-plate dielectric barrier discharge (DBD) was measured by the asynchronous laser-induced fluorescence (LIF) technique and the fluctuation of OH radical density was measured simultaneously to prove that the average OH density can be obtained by the asynchronous LIF technique. The evolution of the average OH density in four different discharge patterns, namely, negative barrier corona discharge, glow discharge, multi glow discharge, and streamer discharge, was studied, and it was found that the average OH density has an observable increase from corona discharge to streamer discharge. The main mechanism of OH production in the four different discharge patterns was analyzed. It was shown that the main mechanism of OH production in negative barrier corona discharge is electron direct collision dissociation, whereas in the other three discharge patterns the He metastable Penning ionization is the main process.

  8. Containerless laser-induced fluorescence study of vaporization and optical properties for sapphire and alumina

    Energy Technology Data Exchange (ETDEWEB)

    Nordine, P.C.; Schiffman, R.A. (Midwest Research Institute, Kansas City, MO (USA) Yale Univ., New Haven, CT (USA))

    1988-09-01

    Evaporation of aluminum oxide was studied from 1,800 to 2,327 K by laser-induced fluorescence (LIF) detection of Al atom vapor over sapphire and alumina spheres that were levitated in an argon gas jet and heated with a continuous wave CO{sub 2} laser. Optical properties were determined from apparent specimen temperatures measured with an optical pyrometer and true temperatures deduced from the LIF intensity versus temperature measurements using the known temperature dependence of the Al atom vapor concentration in equilibrium with Al{sub 2}O{sub 3}. The effects of impurities and dissolved oxygen on the high-temperature optical properties of aluminum oxide were discussed.

  9. Laser-induced fluorescence measurements on plasma science experiments at PPPL

    Energy Technology Data Exchange (ETDEWEB)

    Koepke, Mark

    2011-12-20

    Collaborative research between WVU and PPPL was carried out at WVU for the purpose of incorporating the sophisticated diagnostic technique known as laser-induced fluorescence (LIF) in the Paul-Trap Simulation Experiment (PTSX) at PPPL. WVU assembled a LIF system at WVU, transported it to PPPL, helped make LIF experiments on the PTSX device, participated in PTSX science, and trained PPPL staff in LIF techniques. In summary, WVU refurbished a non-operational LIF system being loaned from University of Maryland to PPPL and, by doing so, provided PPPL with additional diagnostic capability for its PTSX device and other General Plasma Science experiments. WVU students, staff, and faculty will visit PPPL to collaborate on PTSX experiments in the future.

  10. A novel laser-induced fluorescence scheme for Ar-I in a plasma

    Energy Technology Data Exchange (ETDEWEB)

    Short, Zachary D., E-mail: zdshort@mix.wvu.edu; Siddiqui, M. Umair; Henriquez, Miguel F.; McKee, John S.; Scime, Earl E. [Department of Physics and Astronomy, West Virginia University, Morgantown, West Virginia 26506 (United States)

    2016-01-15

    Here we describe a novel infrared laser-induced fluorescence scheme for the 1s{sub 2} state of Ar-I using an 841.052 nm (vacuum) Sacher tunable diode laser oscillator and compare it to an established 667.913 nm (vacuum) 1s{sub 4}-pumping Ar-I LIF scheme using a master oscillator power amplifier laser [A. M. Keesee et al. Rev. Sci. Instrum. 75, 4091 (2004)]. The novel scheme exhibits a significantly greater signal-to-noise ratio for a given injected laser power than the established scheme. We argue that this is caused by less intense spontaneous Ar-I radiation near the LIF emission wavelength for the 1s{sub 2} scheme as compared to the 1s{sub 4} scheme. In addition we present an updated iodine cell spectrum around the 1s{sub 4} LIF scheme pump wavelength.

  11. Quantitative analysis of tetracycline-inducible expression of the green fluorescent protein gene in transgenic chickens.

    Science.gov (United States)

    Koo, Bon Chul; Kwon, Mo Sun; Roh, Ji Yeol; Kim, Minjee; Kim, Jin-Hoi; Kim, Teoan

    2012-01-01

    The use of transgenic farm animals as "bioreactors" to address the growing demand for biopharmaceuticals, both in terms of increased quantity and greater number, represents a key development in the advancement of medical science. However, the potential for detrimental side-effects as a result of uncontrolled constitutive expression of foreign genes in transgenic animals is a well-recognized limitation of such systems. Previously, using a tetracycline-inducible expression system, we demonstrated the induction of expression of a transgene encoding green fluorescent protein (GFP) in transgenic chickens by feeding with doxycycline, a tetracycline derivative; expression of GFP reverted to pre-induction levels when the inducer was removed from the diet. As a proof of principle study, however, quantitative assessment of expression was not possible, as only one G0 and one G1 transgenic chicken was obtained. In the current study, a sufficient number of G2 and G3 transgenic chickens were obtained, and quantification analysis demonstrated up to a 20-fold induction of expression by doxycycline. In addition, stable transmission of the transgene without any apparent genetic modifications was observed through several generations. The use of an inducible expression system that can be regulated by dietary supplementation could help mitigate the physiological disruption that can occur in transgenic animals as a result of uncontrolled constitutive expression of a transgene. Importantly, these results also support the use of the retroviral system for generating transgenic animals with minimal risk in terms of biosafety.

  12. An optical fiber taper fluorescent probe for detection of nitro-explosives based on tetraphenylethylene with aggregation-induced emission

    Science.gov (United States)

    Liu, Fukun; Cui, Minxin; Ma, Jiajun; Zou, Gang; Zhang, Qijin

    2017-07-01

    In this work, we report a novel optical fiber taper fluorescent probe for detection of nitro-explosives. The probe was fabricated by an in-situ photo-plating through evanescent wave and transmitted light initiated thiol-ene ;click; reaction, from which a cross-linked fluorescence porous polymer film was covalently bonded on the surface of the fiber taper. The film exhibits well-organized porous structure due to the presence of polyhedral oligomeric vinylsilsesquioxane moieties, and simultaneously displays strong fluorescence from tetraphenylethylene with aggregation-induced emission property. These two characters make the probe show a remarkable sensitivity, anti-photo-bleaching and a repeatability in detection of TNT and DNT vapors by fluorescence quenching. In addition, the detection is not interfered in the presence of other volatile organic gases.

  13. Determination of the biodiesel content in diesel/biodiesel blends: a method based on fluorescence spectroscopy.

    Science.gov (United States)

    Scherer, Marisa D; Oliveira, Samuel L; Lima, Sandro M; Andrade, Luis H C; Caires, Anderson R L

    2011-05-01

    Blends of biodiesel and diesel are being used increasingly worldwide because of environmental, economic, and social considerations. Several countries use biodiesel blends with different blending limits. Therefore, it is necessary to develop or improve methods to quantify the biodiesel level in a diesel/biodiesel blend, to ensure compliance with legislation. The optical technique based on the absorption of light in the mid-infrared has been successful for this application. However, this method presents some challenges that must be overcome. In this paper, we propose a novel method, based on fluorescence spectroscopy, to determine the biodiesel content in the diesel/biodiesel blend, which allows in loco measurements by using portable systems. The results showed that this method is both practical and more sensitive than the standard optical method. © Springer Science+Business Media, LLC 2011

  14. Detection of polycyclic aromatic hydrocarbons (PAHs) in raw menhaden fish oil using fluorescence spectroscopy: Method development.

    Science.gov (United States)

    Pena, Edwin A; Ridley, Lauren M; Murphy, Wyatt R; Sowa, John R; Bentivegna, Carolyn S

    2015-09-01

    Raw menhaden fish oil was developed for biomonitoring polycyclic aromatic hydrocarbons (PAHs) using fluorescence spectroscopy. Menhaden (Genus Brevoortia) were collected in 2010 and/or 2011 from Delaware Bay, New Jersey, USA; James River, Virginia, USA; Vermillion Bay, Louisiana, USA (VBLA); and Barataria Bay, Louisiana, USA (BBLA). Barataria Bay, Louisiana received heavy oiling from the Deepwater Horizon oil spill. Method development included determining optimal wavelengths for PAH detection, fish oil matrix interferences, and influence of solvent concentration on extraction. Results showed that some fish oils contained high molecular weight PAH-like compounds in addition to other fluorescent compounds such as albumin and vitamin A and vitamin E. None of these naturally occurring compounds interfered with detection of high molecular weight PAHs. However, data suggested that the lipid component of fish oil was altering fluorescence spectra by supporting the formation of PAH excimers. For example, the most intense excitation wavelength for hydroxypyrene shifted from Ex285/Em430 to Ex340/Em430. Comparison of Deepwater Horizon crude oil and fish oil spectra indicated that some fish oils contained crude oil-like PAHs. Using wavelengths of Ex360/Em430, fish oil concentrations were calculated as 3.92 μg/g, 0.61 μg/g, and 0.14 μg/g for a Delaware Bay sample, BBLA 2011, and VBLA 2011, respectively. Overall, these results supported using menhaden fish oil to track PAH exposures spatially and temporally.

  15. An enhanced method of obtaining uniform excitation radiation for fluorescence microscopy

    Science.gov (United States)

    Dimas, Chris F.; Kuta, John J.; Hubert, Manfred

    2003-12-01

    A novel lightsource to provide the excitation radiation for fluorescence microscopy is presented and its performance is compared to the current de factor standard in the field: mercury short arc lamps. This novel light source is remote to the microscope, and the radiation is coupled to the microscope via a liquid lightguide or fiber optic cable using special coupling optics. We present measurements made on some common fluorescent microscopes that show the new light source provides for higher overall optical power delivered to the sample and provides more uniform illumination of the microscopes' field of view in comparison to the standard short arc lamps. Using the definition of the Koehler illumination rules it is shown that the inherent design of the remote source makes it resistant to many non-uniformities and misalignments commonly enountered with the short arc lamp sources; thereby providing for a consistent, uniform irradiance and intensity distribution of the entrance pupil to the microscope. The experimental method used to quantitatively measure the uniformity of the excitation radiation at the microscope's objective plane is also discussed and shown to be far more reliable than other techniques which rely upon fluorescent radiation from synthetic samples placed at the objective plane.

  16. Distribution of aluminum phthalocyanine disulfonate in an oral squamous cell carcinoma model. In vivo fluorescence imaging compared with ex vivo analytical methods

    NARCIS (Netherlands)

    Witjes, MJH; Mank, AJG; Speelman, OC; Posthumus, R; Nooren, CAAM; Nauta, JM; Roodenburg, JLN

    1997-01-01

    Photosensitizer-induced fluorescence is studied as a technique for the detection of cancer, Therefore we investigated the ability of a photosensitizer, aluminum phthalocyanine disulfonate (AlPcS2), to localize in tumor tissue. In vivo endoscopic fluorescence imaging, fluorescence microscopy, convent

  17. A general method for site specific fluorescent labeling of recombinant chemokines.

    Directory of Open Access Journals (Sweden)

    Tetsuya Kawamura

    Full Text Available Chemokines control cell migration in many contexts including development, homeostasis, immune surveillance and inflammation. They are also involved in a wide range of pathological conditions ranging from inflammatory diseases and cancer, to HIV. Chemokines function by interacting with two types of receptors: G protein-coupled receptors on the responding cells, which transduce signaling pathways associated with cell migration and activation, and glycosaminoglycans on cell surfaces and the extracellular matrix which organize and present some chemokines on immobilized surface gradients. To probe these interactions, imaging methods and fluorescence-based assays are becoming increasingly desired. Herein, a method for site-specific fluorescence labeling of recombinant chemokines is described. It capitalizes on previously reported 11-12 amino acid tags and phosphopantetheinyl transferase enzymes to install a fluorophore of choice onto a specific serine within the tag through a coenzyme A-fluorophore conjugate. The generality of the method is suggested by our success in labeling several chemokines (CXCL12, CCL2, CCL21 and mutants thereof and visualizing them bound to chemokine receptors and glycosaminoglycans. CXCL12 and CCL2 showed the expected co-localization on the surface of cells with their respective receptors CXCR4 and CCR2 at 4 °C, and co-internalization with their receptors at 37 °C. By contrast, CCL21 showed the presence of large discrete puncta that were dependent on the presence of both CCR7 and glycosaminoglycans as co-receptors. These data demonstrate the utility of this labeling approach for the detection of chemokine interactions with GAGs and receptors, which can vary in a chemokine-specific manner as shown here. For some applications, the small size of the fluorescent adduct may prove advantageous compared to other methods (e.g. antibody labeling, GFP fusion by minimally perturbing native interactions. Other advantages of the method

  18. Simultaneous determination of amino acids in tea leaves by micellar electrokinetic chromatography with laser-induced fluorescence detection.

    Science.gov (United States)

    Yan, Jin; Cai, Yuanli; Wang, Yufei; Lin, Xia; Li, Hui

    2014-01-15

    A rapid and effective method of micellar electrokinetic chromatography with laser-induced fluorescence detection was developed for the simultaneous determination of amino acids in tea leaves. Pre-column derivatization of the analytes used 4-chloro-7-nitrobenzofurazan (NDB-Cl). Optimal separation was achieved at +20kV using an uncoated fused silica capillary (40.0cm effective length, 50.2cm total length, 75μm internal diameter), as well as 20mM sodium borate (pH 8.5), 20mM Brij 35, and acetonitrile 10% (v/v) as running buffers. Within 11min, 15 amino acids were separated completely. The optimized method demonstrated good linearity (r(2)⩾0.9990), precision (⩽6.65%), accuracy (85.50-112.74%), and sensitivity (0.1ng/mL-100ng/mL). The method successfully determined the quantity of amino acids in five different tea leaves; furthermore, theanine was identified as the most abundant amino acid in teas. The proposed method showed great potential in further investigations on the biofunctions of different tea samples.

  19. Data preprocessing method for fluorescence molecular tomography using a priori information provided by CT.

    Science.gov (United States)

    Fu, Jianwei; Yang, Xiaoquan; Meng, Yuanzheng; Luo, Qingming; Gong, Hui

    2012-01-01

    The combined system of micro-CT and fluorescence molecular tomography (FMT) offers a new tool to provide anatomical and functional information of small animals in a single study. To take advantages of the combined system, a data preprocessing method is proposed to extract the valid data for FMT reconstruction algorithms using a priori information provided by CT. The boundary information of the animal and animal holder is extracted from reconstructed CT volume data. A ray tracing method is used to trace the path of the excitation beam, calculate the locations and directions of the optional sources and determine whether the optional sources are valid. To accurately calculate the projections of the detectors on optical images and judge their validity, a combination of perspective projection and inverse ray tracing method are adopted to offer optimal performance. The imaging performance of the combined system with the presented method is validated through experimental rat imaging.

  20. Radiation induced chromatin conformation changes analysed by fluorescent localization microscopy, statistical physics, and graph theory.

    Science.gov (United States)

    Zhang, Yang; Máté, Gabriell; Müller, Patrick; Hillebrandt, Sabina; Krufczik, Matthias; Bach, Margund; Kaufmann, Rainer; Hausmann, Michael; Heermann, Dieter W

    2015-01-01

    It has been well established that the architecture of chromatin in cell nuclei is not random but functionally correlated. Chromatin damage caused by ionizing radiation raises complex repair machineries. This is accompanied by local chromatin rearrangements and structural changes which may for instance improve the accessibility of damaged sites for repair protein complexes. Using stably transfected HeLa cells expressing either green fluorescent protein (GFP) labelled histone H2B or yellow fluorescent protein (YFP) labelled histone H2A, we investigated the positioning of individual histone proteins in cell nuclei by means of high resolution localization microscopy (Spectral Position Determination Microscopy = SPDM). The cells were exposed to ionizing radiation of different doses and aliquots were fixed after different repair times for SPDM imaging. In addition to the repair dependent histone protein pattern, the positioning of antibodies specific for heterochromatin and euchromatin was separately recorded by SPDM. The present paper aims to provide a quantitative description of structural changes of chromatin after irradiation and during repair. It introduces a novel approach to analyse SPDM images by means of statistical physics and graph theory. The method is based on the calculation of the radial distribution functions as well as edge length distributions for graphs defined by a triangulation of the marker positions. The obtained results show that through the cell nucleus the different chromatin re-arrangements as detected by the fluorescent nucleosomal pattern average themselves. In contrast heterochromatic regions alone indicate a relaxation after radiation exposure and re-condensation during repair whereas euchromatin seemed to be unaffected or behave contrarily. SPDM in combination with the analysis techniques applied allows the systematic elucidation of chromatin re-arrangements after irradiation and during repair, if selected sub-regions of nuclei are

  1. Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Hui Su

    2001-05-25

    Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, the author introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, they demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm{sup 2} for 40-{micro}m wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection. In the second part of this dissertation, the author used laser-induced native fluorescence coupled with capillary electrophoresis (LINF-CE) and microscope imaging to study the single cell degranulation. On the basis of good temporal correlation with events observed through an optical microscope, they have identified individual peaks in the fluorescence electropherograms as serotonin released from the granular core on contact with the surrounding fluid.

  2. One- and two-photon induced fluorescence spectroscopy enabling the detection of localized aflatoxin contamination in individual maize kernels

    Science.gov (United States)

    Smeesters, L.; Meulebroeck, W.; Raeymaekers, S.; Thienpont, H.

    2016-04-01

    The presence of carcinogenic aflatoxins in food and feed products is a major worldwide problem. To date, the aflatoxin contamination can only be detected by the use of destructive sample-based chemical analyses. Therefore, we developed an optical setup able to detect the localized aflatoxin contamination in individual maize kernels, on the basis of one- and two- photon induced fluorescence spectroscopy. Our developed optical configuration comprises a tunable titanium-sapphire laser (710nm-830nm) in combination with second harmonic wavelength generation (355nm-415nm), enabling the measurement of both one- and two-photon induced fluorescence spectra. Moreover, an accurate scanning of the kernel's surface was induced by the use of automated translation stages, allowing to study the localized maize contamination. First, the operation of the setup is validated by the characterization of pure aflatoxin B1 powder. Second, the fluorescence spectra of healthy (maize kernels (>70ppb aflatoxin B1) were measured, after excitation with 365nm, 730nm, 750nm and 780nm. For both the one- and two- photon induced fluorescence processes, the presence of the aflatoxin inside the contaminated maize kernels influenced the intrinsic fluorescence signals. Based on the fluorescence spectrum between 400nm and 550nm, we defined a detection criterion to identify the contaminated maize kernels. Furthermore, we demonstrate the sensing of the localized contamination level, indicating both contaminated maize kernels with a high contamination level in a limited surface area (as small as 1mm2) as with a lower contamination spread over a large surface area (up to 20mm2). As a result, our developed measurement methodology allows the identification of the localized aflatoxin contamination, paving the way to the non-destructive, real-time and high-sensitive industrial scanning-based detection of aflatoxins in food products.

  3. Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Su, Hui [Iowa State Univ., Ames, IA (United States)

    2001-01-01

    Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, the author introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, they demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm2 for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection. In the second part of this dissertation, the author used laser-induced native fluorescence coupled with capillary electrophoresis (LINF-CE) and microscope imaging to study the single cell degranulation. On the basis of good temporal correlation with events observed through an optical microscope, they have identified individual peaks in the fluorescence electropherograms as serotonin released from the granular core on contact with the surrounding fluid.

  4. Noninvasive control of dental calculus removal: qualification of two fluorescence methods

    Science.gov (United States)

    Gonchukov, S.; Sukhinina, A.; Bakhmutov, D.; Biryukova, T.

    2013-02-01

    The main condition of periodontitis prevention is the full calculus removal from the teeth surface. This procedure should be fulfilled without harming adjacent unaffected tooth tissues. Nevertheless the problem of sensitive and precise estimating of tooth-calculus interface exists and potential risk of hard tissue damage remains. In this work it was shown that fluorescence diagnostics during calculus removal can be successfully used for precise noninvasive detection of calculus-tooth interface. In so doing the simple implementation of this method free from the necessity of spectrometer using can be employed. Such a simple implementation of calculus detection set-up can be aggregated with the devices of calculus removing.

  5. Indirect detection of superoxide in RAW 264.7 macrophage cells using microchip electrophoresis coupled to laser-induced fluorescence.

    Science.gov (United States)

    de Campos, Richard P S; Siegel, Joseph M; Fresta, Claudia G; Caruso, Giuseppe; da Silva, José A F; Lunte, Susan M

    2015-09-01

    Superoxide, a naturally produced reactive oxygen species (ROS) in the human body, is involved in many pathological and physiological signaling processes. However, if superoxide formation is left unregulated, overproduction can lead to oxidative damage to important biomolecules, such as DNA, lipids, and proteins. Superoxide can also lead to the formation of peroxynitrite, an extremely hazardous substance, through its reaction with endogenously produced nitric oxide. Despite its importance, quantitative information regarding superoxide production is difficult to obtain due to its high reactivity and low concentrations in vivo. MitoHE, a fluorescent probe that specifically reacts with superoxide, was used in conjunction with microchip electrophoresis (ME) and laser-induced fluorescence (LIF) detection to investigate changes in superoxide production by RAW 264.7 macrophage cells following stimulation with phorbol 12-myristate 13-acetate (PMA). Stimulation was performed in the presence and absence of the superoxide dismutase (SOD) inhibitors, diethyldithiocarbamate (DDC) and 2-metoxyestradiol (2-ME). The addition of these inhibitors resulted in an increase in the amount of superoxide specific product (2-OH-MitoE(+)) from 0.08 ± 0.01 fmol (0.17 ± 0.03 mM) in native cells to 1.26 ± 0.06 fmol (2.5 ± 0.1 mM) after PMA treatment. This corresponds to an approximately 15-fold increase in intracellular concentration per cell. Furthermore, the addition of 3-morpholino-sydnonimine (SIN-1) to the cells during incubation resulted in the production of 0.061 ± 0.006 fmol (0.12 ± 0.01 mM) of 2-OH-MitoE(+) per cell on average. These results demonstrate that indirect superoxide detection coupled with the use of SOD inhibitors and a separation method is a viable method to discriminate the 2-OH-MitoE(+) signal from possible interferences.

  6. Fluorescence diagnosis in tissue injury

    Science.gov (United States)

    Maciel, Vitória H.; Ferreira, Juliana; Bagnato, Vanderlei S.

    2009-06-01

    Background and Objectives: The paper aim was to evaluate the efficacy of the fluorescence spectroscopy in the detection of UV-induced skin change of Wistar rats. Study Design/ Materials and Methods: In a group male Wistar rats, the skin damage was produced by an UV-C lamp, periodically monitored using the laser-induced fluorescence, until complete healing process. After determining a characteristic emission band present in the fluorescence spectra of the induced injuries, the amplitude band monitoring allowed the follow up on the injury and the recovery. Results: We observed the appearance of two new emission bands more evident at the injury spectra when compared to the spectrums from normal non-exposed tissue. Following such spectral bands was possible to observe the establishment and recovery. Conclusions: The fluorescence spectroscopy is a promising technique in distinguishing between normal and UV induced skin change helping the evaluation of changes which are irreversible cancer tissue characteristics.

  7. Teaching about photosynthesis with simple equipment: analysis of light-induced changes in fluorescence and reflectance of plant leaves.

    Science.gov (United States)

    Björn, Lars Olof; Li, Shaoshan

    2013-10-01

    Solar energy absorbed by plants results in either reflection or absorption. The latter results in photosynthesis, fluorescence, or heat. Measurements of fluorescence changes have been used for monitoring processes associated with photosynthesis. A simple method to follow changes in leaf fluorescence and leaf reflectance associated with nonphotochemical quenching and light acclimation of leaves is described. The main equipment needed consists of a green-light emitting laser pointer, a digital camera, and a personal computer equipped with the camera acquisition software and the programs ImageJ and Excel. Otherwise, only commonly available cheap materials are required.

  8. Functionalization of embedded thiol-ene waveguides for evanescent wave-induced fluorescence detection in a microfluidic device

    DEFF Research Database (Denmark)

    Feidenhans, Nikolaj A.; Jensen, Thomas Glasdam; Lafleur, Josiane P.;

    2013-01-01

    We demonstrate the use of functional surface groups inherently present on off-stoichiometric thiol-ene polymers, for site-specific immobilization of biomolecules and detection by evanescent wave-induced fluorescence. An optofluidic chip featuring an embedded thiol-ene waveguide was selectively fu...

  9. A Quantitative Theoretical Framework For Protein-Induced Fluorescence Enhancement-Förster-Type Resonance Energy Transfer (PIFE-FRET)

    NARCIS (Netherlands)

    Lerner, Eitan; Ploetz, Evelyn; Hohlbein, Johannes; Cordes, Thorben; Weiss, Shimon

    2016-01-01

    Single-molecule, protein-induced fluorescence enhancement (PIFE) serves as a molecular ruler at molecular distances inaccessible to other spectroscopic rulers such as Förster-type resonance energy transfer (FRET) or photoinduced electron transfer. In order to provide two simultaneous measurements of

  10. A Quantitative Theoretical Framework for Protein-Induced Fluorescence Enhancement-Förster-Type Resonance Energy Transfer (PIFE-FRET)

    NARCIS (Netherlands)

    Lerner, Eitan; Ploetz, Evelyn; Hohlbein, Johannes; Cordes, Thorben; Weiss, Shimon

    2016-01-01

    Single-molecule, protein-induced fluorescence enhancement (PIFE) serves as a molecular ruler at molecular distances inaccessible to other spectroscopic rulers such as Förster-type resonance energy transfer (FRET) or photoinduced electron transfer. In order to provide two simultaneous measurements

  11. Fluorescence Aggregation-Caused Quenching versus Aggregation-Induced Emission: A Visual Teaching Technology for Undergraduate Chemistry Students

    Science.gov (United States)

    Ma, Xiaofeng; Sun, Rui; Cheng, Jinghui; Liu, Jiaoyan; Gou, Fei; Xiang, Haifeng; Zhou, Xiangge

    2016-01-01

    A laboratory experiment visually exploring two opposite basic principles of fluorescence of aggregation-caused quenching (ACQ) and aggregation-induced emission (AIE) is demonstrated. The students would prepared two salicylaldehyde-based Schiff bases through a simple one-pot condensation reaction of one equiv of 1,2-diamine with 2 equiv of…

  12. Spaceborne Sun-Induced Vegetation Fluorescence Time Series from 2007 to 2015 Evaluated with Australian Flux Tower Measurements

    NARCIS (Netherlands)

    Sanders, Abram; Verstraeten, Willem; Kooreman, Maurits; Leth, Van Thomas; Beringer, Jason; Joiner, Joanna

    2016-01-01

    A global, monthly averaged time series of Sun-induced Fluorescence (SiF), spanning January 2007 to June 2015, was derived from Metop-A Global Ozone Monitoring Experiment 2 (GOME-2) spectral measurements. Far-red SiF was retrieved using the filling-in of deep solar Fraunhofer lines and atmospheric ab

  13. Native Fluorescence Detection Methods and Detectors for Naphthalene and/or Other Volatile Organic Compound Vapors

    Science.gov (United States)

    Hug, William F. (Inventor); Bhartia, Rohit (Inventor); Reid, Ray D. (Inventor); Lane, Arthur L. (Inventor)

    2014-01-01

    Naphthalene, benzene, toluene, xylene, and other volatile organic compounds have been identified as serious health hazards. This is especially true for personnel working with JP8 jet fuel and other fuels containing naphthalene as well as other hazardous volatile organic compounds (VOCs). Embodiments of the invention are directed to methods and apparatus for near-real-time in-situ detection and accumulated dose measurement of exposure to naphthalene vapor and other hazardous gaseous VOCs. The methods and apparatus employ excitation of fluorophors native or endogenous to compounds of interest using light sources emitting in the ultraviolet below 300 nm and measurement of native fluorescence emissions in distinct wavebands above the excitation wavelength. The apparatus of some embodiments are cell-phone-sized sensor/dosimeter "badges" to be worn by personnel potentially exposed to naphthalene or other hazardous VOCs. The badge sensor of some embodiments provides both real time detection and data logging of exposure to naphthalene or other VOCs of interest from which both instantaneous and accumulated dose can be determined. The badges employ a new native fluorescence based detection method to identify and differentiate VOCs. The particular focus of some embodiments are the detection and identification of naphthalene while other embodiments are directed to detection and identification of other VOCs like aromatic hydrocarbons such as benzene, toluene, and xylene.

  14. A virtual source pattern method for fluorescence tomography with structured light.

    Science.gov (United States)

    Ducros, Nicolas; D'Andrea, Cosimo; Bassi, Andrea; Valentini, Gianluca; Arridge, Simon

    2012-06-21

    In order to reduce both acquisition and reconstruction times, illumination and detection in fluorescence diffuse optical tomography (FDOT) have recently evolved from a point-based to a pattern-based approach. The use of structured illumination, offering the ability to project any pattern of light onto the object, associated with the compression of the acquired fluorescence images has paved the way for a new generation of fast reconstruction algorithms for FDOT. However, the choice of the most appropriate set of source patterns is still an open problem. Here, the use of typical source patterns is investigated on experimental data. Reconstructions of similar qualities are obtained for the different types of source patterns. We found that the performances of structured illumination are limited by the required positivity of the source patterns. To alleviate this problem, we introduce a novel method, namely the virtual source pattern method, which allows for considering any kind of patterns, e.g., with negative and complex intensities. This new method provides a significant increase of the contrast of the reconstruction and also a reduction of the reconstruction error, especially when virtual wavelet source patterns are considered.

  15. Simultaneous determination of vigabatrin and amino acid neurotransmitters in brain microdialysates by capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Benturquia, Nadia; Parrot, Sandrine; Sauvinet, Valérie; Renaud, Bernard; Denoroy, Luc

    2004-07-01

    Capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD) coupled to in vivo microdialysis sampling was used in order to monitor simultaneously a drug and several neurotransmitters in the brain extracellular fluid. Determination of the antiepileptic drug vigabatrin and the amino acid neurotransmitters glutamate (Glu), l-aspartate (l-Asp) and gamma-aminobutyric acid (GABA) was performed on low-concentration samples which were derivatized with naphthalene-2,3-dicarboxaldehyde (NDA) and separated using a pH 9.2 75 mM sodium borate running buffer containing 60 mM sodium dodecyl sulfate (SDS) and 5mM hydroxypropyl-beta-cyclodextrin (HP-beta-CD). Glu, l-Asp and vigabatrin derivatized at a concentration of 1.0 x 10(-9) M, and GABA derivatized at a concentration of 5.0 x 10(-9) M, produced peaks with signal-to-noise ratios of 8:1, 8:1, 4:1 and 5:1, respectively. The nature of the neurotransmitter peaks found in rat brain microdialysates was confirmed by both electrophoretic and pharmacological validations. This method was used for monitoring vigabatrin and amino acid neurotransmitters in microdialysates from the rat striatum during intracerebral infusion of the drug and revealed rapid vigabatrin-induced changes in GABA and Glu levels. This original application of CE-LIFD coupled to microdialysis represents a powerful tool for pharmacokinetic/pharmacodynamic investigations.

  16. Absorption into fluorescence. A method to sense biologically relevant gas molecules.

    Science.gov (United States)

    Strianese, Maria; Varriale, Antonio; Staiano, Maria; Pellecchia, Claudio; D'Auria, Sabato

    2011-01-01

    In this work we present an innovative optical sensing methodology based on the use of biomolecules as molecular gating nano-systems. Here, as an example, we report on the detection of analytes related to climate change. In particular, we focused our attention on the detection of nitric oxide (NO) and oxygen (O2). Our methodology builds on the possibility of modulating the excitation intensity of a fluorescent probe used as a transducer and a sensor molecule whose absorption is strongly affected by the binding of an analyte of interest used as a filter. The two simple conditions that have to be fulfilled for the method to work are: (a) the absorption spectrum of the sensor placed inside the cuvette, and acting as the recognition element for the analyte of interest, should strongly change upon the binding of the analyte and (b) the fluorescence dye transducer should exhibit an excitation band which overlaps with one or more absorption bands of the sensor. The absorption band of the sensor affected by the binding of the specific analyte should overlap with the excitation band of the transducer. The high sensitivity of fluorescence detection combined with the use of proteins as highly selective sensors makes this method a powerful basis for the development of a new generation of analytical assays. Proof-of-principle results showing that cytochrome c peroxidase (CcP) for NO detection and myoglobin (Mb) for O2 detection can be successfully used by exploiting our new methodology are reported. The proposed technology can be easily expanded to the determination of different target analytes.

  17. Analysis of the anti-Parkinson drug pramipexole in human urine by capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Musenga, Alessandro; Kenndler, Ernst; Morganti, Emanuele; Rasi, Fabrizio; Raggi, Maria Augusta

    2008-09-19

    A sensitive method based on capillary electrophoresis with laser-induced fluorescence detection has been developed for the analysis of the non-ergoline dopamine agonist pramipexole in human urine. Separation was carried out in uncoated fused silica capillaries (75microm internal diameter, 75.0 and 60.0cm total and effective length, respectively), with a background electrolyte composed of borate buffer (50mM, pH 10.3), tetrabutylammonium bromide (30mM), and acetone (15%, v/v). Applying a 20kV voltage, the electrophoretic run is completed within 12min. A sample pre-treatment procedure based on liquid/liquid extraction with ethyl acetate, followed by derivatisation of pramipexole with fluorescein isothiocyanate at pH 9, allows the complete removal of biological interferences, with extraction yields always higher than 94.5%. Method validation gave good linearity (r(2)=0.9992) in the 25.0-1000ngmL(-1) range; limit of detection and limit of quantitation were 10.0 and 25.0ngmL(-1), respectively; precision was 90.0. The method was applied to the analysis of urine samples from patients undergoing therapy with pramipexole.

  18. Remote measurement of photosynthetic efficiency using laser induced fluorescence transient (LIFT) technique.

    Science.gov (United States)

    Pieruschka, R.; Rascher, U.; Klimov, D.; Kolber, Z. S.; Berry, J. A.

    2007-12-01

    An understanding of spatial and temporal diversity of photosynthetic processes, water and energy exchange of complex plant canopies is essential for carbon and climate models. Remote sensing from space or aircraft platforms provides the only practical way to characterize the vast extent of plant canopies around the globe, but the basis for relating physiological processes to remote sensing is still largely theoretical. Experiments that bridge this gap are needed. Chlorophyll fluorescence measurements have been widely applied to quantify photosynthetic efficiency and non- photochemical energy dissipation non-destructively in photosynthetically active organisms. The most commonly used Pulse Amplitude Modulated (PAM) technique provides a saturating light pulse and is not practical at the canopy scale. We report here on a recently developed technique, Laser Induced Fluorescence Transient (LIFT), capable of remote measurement of photosynthetic efficiency of selected leaves at a distance of up to 50 m and we present here continuous studies on plans growing under natural conditions during the beginning of the winter season and the onset of summer drought in this Mediterranean climate. i) Lichens showed a strong diurnal variation in photosynthetic efficiency which correlated with relative humidity; ii) Photosynthetic efficiency of annual grass decreased with progressing drought stress; iii) An oak canopy showed very little variation of quantum yield from leaf out in spring to summer; iv) The combined effect of low temperature and high light intensity during an early winter strongly reduced the photosynthetic efficiency of four different species in response to chilling stress. These measures with the LIFT correlated well with (more limited) sampling by PAM fluoromentry and gas exchange. The ability to make continuous, automatic and remote measurements of photosynthetic efficiency of leaves with the LIFT provides a new approach for studying the heterogeneity of

  19. Multispectral imaging system based on laser-induced fluorescence for security applications

    Science.gov (United States)

    Caneve, L.; Colao, F.; Del Franco, M.; Palucci, A.; Pistilli, M.; Spizzichino, V.

    2016-10-01

    The development of portable sensors for fast screening of crime scenes is required to reduce the number of evidences useful to be collected, optimizing time and resources. Laser based spectroscopic techniques are good candidates to this scope due to their capability to operate in field, in remote and rapid way. In this work, the prototype of a multispectral imaging LIF (Laser Induced Fluorescence) system able to detect evidence of different materials on large very crowded and confusing areas at distances up to some tens of meters will be presented. Data collected as both 2D fluorescence images and LIF spectra are suitable to the identification and the localization of the materials of interest. A reduced scan time, preserving at the same time the accuracy of the results, has been taken into account as a main requirement in the system design. An excimer laser with high energy and repetition rate coupled to a gated high sensitivity ICCD assures very good performances for this purpose. Effort has been devoted to speed up the data processing. The system has been tested in outdoor and indoor real scenarios and some results will be reported. Evidence of the plastics polypropylene (PP) and polyethilene (PE) and polyester have been identified and their localization on the examined scenes has been highlighted through the data processing. By suitable emission bands, the instrument can be used for the rapid detection of other material classes (i.e. textiles, woods, varnishes). The activities of this work have been supported by the EU-FP7 FORLAB project (Forensic Laboratory for in-situ evidence analysis in a post blast scenario).

  20. Ablation plume structure and dynamics in ambient gas observed by laser-induced fluorescence imaging spectroscopy

    Energy Technology Data Exchange (ETDEWEB)

    Miyabe, M.; Oba, M.; Iimura, H.; Akaoka, K.; Khumaeni, A.; Kato, M.; Wakaida, I.

    2015-08-01

    The dynamic behavior of an ablation plume in ambient gas has been investigated by laser-induced fluorescence imaging spectroscopy. The second harmonic beam from an Nd:YAG laser (0.5–6 J/cm{sup 2}) was focused on a sintered oxide pellet or a metal chip of gadolinium. The produced plume was subsequently intersected with a sheet-shaped UV beam from a dye laser so that time-resolved fluorescence images were acquired with an intensified CCD camera at various delay times. The obtained cross-sectional images of the plume indicate that the ablated ground state atoms and ions of gadolinium accumulate in a hemispherical contact layer between the plume and the ambient gas, and a cavity containing a smaller density of ablated species is formed near the center of the plume. At earlier expansion stage, another luminous component also expands in the cavity so that it coalesces into the hemispherical layer. The splitting and coalescence for atomic plume occur later than those for ionic plume. Furthermore, the hemispherical layer of neutral atoms appears later than that of ions; however, the locations of the layers are nearly identical. This coincidence of the appearance locations of the layers strongly suggests that the neutral atoms in the hemispherical layer are produced as a consequence of three-body recombination of ions through collisions with gas atoms. The obtained knowledge regarding plume expansion dynamics and detailed plume structure is useful for optimizing the experimental conditions for ablation-based spectroscopic analysis. - Highlights: • Ablated ground-state species accumulated in a thin hemispherical boundary layer • Inside the layer, a cavity containing a small density of ablated species was formed. • The hemispherical layers of atoms and ions appeared at a nearly identical location. • The measured intensity peak variation was in good agreement with a model prediction. • We ascribed the dominant process for forming the layer to a three

  1. Laser Induced Fluorescence Spectroscopy of Neutral and Ionized Polycyclic Aromatic Hydrocarbons in a Cosmic Simulation Chamber

    Science.gov (United States)

    Bejaoui, Salma; Salama, Farid

    2015-08-01

    Polycyclic aromatic hydrocarbon (PAH) molecules are considered the best carriers to account for the ubiquitous infrared emission bands. PAHs have also been proposed as candidates to explain the diffuse interstellar bands (DIBs), a series of absorption features seen on the interstellar extinction curve and are plausible carriers for the extended red emission (ERE), a photoluminescent process associated with a wide variety of interstellar environments. Extensive efforts have been devoted over the past two decades to characterize the physical and chemical properties of PAH molecules and ions in space. Absorption spectra of PAH molecules and ions trapped in solid matrices have been compared to the DIBs [1, 2]. Absorption spectra of several cold, isolated gas-phase PAHs have also been measured under experimental conditions that mimic the interstellar conditions [see 3 for a review]. The purpose of this study is to provide a new dimension to the existing spectroscopic database of neutral and single ionized PAHs that is largely based on absorption spectra by adding emission spectroscopy data. The measurements are based on the laser-induced fluorescence (LIF) technique [4] and are performed with the Pulsed Discharge Nozzle (PDN) of the COSmIC laboratory facility at NASA Ames laboratory. The PDN generates plasma in a free supersonic jet expansion to simulate the physical and the chemical conditions in interstellar environments. We focus, here, on the fluorescence spectra of large neutral PAHs and their cations where there is a lack of fluorescence spectroscopy data. The astronomical implications of the data (e.g., ERE) are examinedReferences[1] F. Salama, E. Bakes, L.J. Allamandola, A.G.G.M. Tielens, Astrophys. J., 458 (1996) p.621[2] F. Salama, The ISO Revolution, EDP Sciences, Les Ulis, France (1999) p.65[3] Salama F., In Organic Matter in Space, IAU Symposium 251, Kwok & Sandford Eds.Cambridge University Press,4, S251,(2008), p. 357 (2008) and references therein.[4

  2. Full-direct method for imaging pharmacokinetic parameters in dynamic fluorescence molecular tomography

    Energy Technology Data Exchange (ETDEWEB)

    Zhang, Guanglei, E-mail: guangleizhang@bjtu.edu.cn [Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084 (China); Department of Biomedical Engineering, School of Computer and Information Technology, Beijing Jiaotong University, Beijing 100044 (China); Pu, Huangsheng; Liu, Fei; Bai, Jing [Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084 (China); He, Wei [China Institute of Sport Science, Beijing 100061 (China); Luo, Jianwen, E-mail: luo-jianwen@tsinghua.edu.cn [Department of Biomedical Engineering, School of Medicine, Tsinghua University, Beijing 100084 (China); Center for Biomedical Imaging Research, School of Medicine, Tsinghua University, Beijing 100084 (China)

    2015-02-23

    Images of pharmacokinetic parameters (also known as parametric images) in dynamic fluorescence molecular tomography (FMT) can provide three-dimensional metabolic information for biological studies and drug development. However, the ill-posed nature of FMT and the high temporal variation of fluorophore concentration together make it difficult to obtain accurate parametric images in small animals in vivo. In this letter, we present a method to directly reconstruct the parametric images from the boundary measurements based on hybrid FMT/X-ray computed tomography (XCT) system. This method can not only utilize structural priors obtained from the XCT system to mitigate the ill-posedness of FMT but also make full use of the temporal correlations of boundary measurements to model the high temporal variation of fluorophore concentration. The results of numerical simulation and mouse experiment demonstrate that the proposed method leads to significant improvements in the reconstruction quality of parametric images.

  3. Full-direct method for imaging pharmacokinetic parameters in dynamic fluorescence molecular tomography

    Science.gov (United States)

    Zhang, Guanglei; Pu, Huangsheng; He, Wei; Liu, Fei; Luo, Jianwen; Bai, Jing

    2015-02-01

    Images of pharmacokinetic parameters (also known as parametric images) in dynamic fluorescence molecular tomography (FMT) can provide three-dimensional metabolic information for biological studies and drug development. However, the ill-posed nature of FMT and the high temporal variation of fluorophore concentration together make it difficult to obtain accurate parametric images in small animals in vivo. In this letter, we present a method to directly reconstruct the parametric images from the boundary measurements based on hybrid FMT/X-ray computed tomography (XCT) system. This method can not only utilize structural priors obtained from the XCT system to mitigate the ill-posedness of FMT but also make full use of the temporal correlations of boundary measurements to model the high temporal variation of fluorophore concentration. The results of numerical simulation and mouse experiment demonstrate that the proposed method leads to significant improvements in the reconstruction quality of parametric images.

  4. Observation of hydrocarbon generation and migration of highly-matured carbonates by means of laser-induced fluorescence microscopy

    Institute of Scientific and Technical Information of China (English)

    2000-01-01

    Some important information on hydrocarbon generation, inclusion and migration in highly-matured carbonates of lower Palaeozoic age from the Ordos Basin and Tarim Basin hasbeen analyzed by a newly-combined laser-induced fluorescence microscope (LFM) designed by our laboratory. The following information has been obtained from the lower Ordovician lamellar carbonates with equivalent vitrinite reflectance (Ro) as high as 1.6%-1.7% and residual TOC of 0.14%-0.35% from the Ordos Basin: wide occurrences of oil and source macerals with strong fluorescence, including G. Prisca alginite, lamalginite, telalginite and algae-detrinite; fluorescing asphalt among mineral crystals; some groundmass and spheroid-like reservoir bitumen with high maturation levels in the pores of dolomites. Various kinds of fluorescing organic inclusions and asphalt have been found in the carbonates, calcareous shales and silt-shales with high maturation levels from the Cambrian-Ordovician strata in the Tarim Basin. All this helps us find and evaluate significant and excellent source rocks for large-and middle-scale gas fields. The net and micro-net systems for hydrocarbon generation, expulsion and migration in carbonates have been revealed by the highly-powered laser-induced fluorescence microscopy.

  5. Characterization of type I, II, III, IV, and V collagens by time-resolved laser-induced fluorescence spectroscopy

    Science.gov (United States)

    Marcu, Laura; Cohen, David; Maarek, Jean-Michel I.; Grundfest, Warren S.

    2000-04-01

    The relative proportions of genetically distinct collagen types in connective tissues vary with tissue type and change during disease progression, development, wound healing, aging. This study aims to 1) characterize the spectro- temporal fluorescence emission of fiber different types of collagen and 2) assess the ability of time-resolved laser- induced fluorescence spectroscopy to distinguish between collagen types. Fluorescence emission of commercially available purified samples was induced with nitrogen laser excitation pulses and detected with a MCP-PMT connected to a digital storage oscilloscope. The recorded time-resolved emission spectra displayed distinct fluorescence emission characteristics for each collagen type. The time domain information complemented the spectral domain intensity data for improved discrimination between different collagen types. Our results reveal that analysis of the fluorescence emission can be used to characterize different species of collagen. Also, the results suggest that time-resolved spectroscopy can be used for monitoring of connective tissue matrix composition changes due to various pathological and non-pathological conditions.

  6. Improved maximum entropy method for the analysis of fluorescence spectroscopy data: evaluating zero-time shift and assessing its effect on the determination of fluorescence lifetimes.

    Science.gov (United States)

    Esposito, Rosario; Mensitieri, Giuseppe; de Nicola, Sergio

    2015-12-21

    A new algorithm based on the Maximum Entropy Method (MEM) is proposed for recovering both the lifetime distribution and the zero-time shift from time-resolved fluorescence decay intensities. The developed algorithm allows the analysis of complex time decays through an iterative scheme based on entropy maximization and the Brent method to determine the minimum of the reduced chi-squared value as a function of the zero-time shift. The accuracy of this algorithm has been assessed through comparisons with simulated fluorescence decays both of multi-exponential and broad lifetime distributions for different values of the zero-time shift. The method is capable of recovering the zero-time shift with an accuracy greater than 0.2% over a time range of 2000 ps. The center and the width of the lifetime distributions are retrieved with relative discrepancies that are lower than 0.1% and 1% for the multi-exponential and continuous lifetime distributions, respectively. The MEM algorithm is experimentally validated by applying the method to fluorescence measurements of the time decays of the flavin adenine dinucleotide (FAD).

  7. Laser-induced fluorescence measurement of the oil film thickness in an internal combustion engine

    Science.gov (United States)

    Ostroski, Greg M.; Ghandhi, Jaal B.

    1997-11-01

    The use of a fluorescent dopant molecule to enhance the natural fluorescence of motor oils, and allow quantitative determination of temperature and film thickens in internal combustion engines has been investigated. Measurement of the fluorescence as a function of temperature were made with neat Mobil 1, and solutions of the dopant BTBP in mineral oil and Mobil 1. The fluorescence yield of neat Mobil 1 was found to vary by 30 percent over the temperature range explored, but the spectral characteristics, as measured with bandpass filters, were unaffected by temperature. The BTBP fluorescence was found to increase significantly with temperature, and it was found the narrower regions in the spectrum increased proportionally more than the fluorescence collected over the entire spectrum, allowing a determination of temperature to be made which can then be used to correct for the change in fluorescence yield. Solutions in Mobil 1 showed a smaller increase than that observed in mineral oil.

  8. A LED-based method for monitoring NAD(P)H and FAD fluorescence in cell cultures and brain slices.

    Science.gov (United States)

    Rösner, Jörg; Liotta, Agustin; Schmitz, Dietmar; Heinemann, Uwe; Kovács, Richard

    2013-01-30

    Nicotinamide- and flavine-adenine-dinucleotides (NAD(P)H and FADH₂) are electron carriers involved in cellular energy metabolism and in a multitude of enzymatic processes. As reduced NAD(P)H and oxidised FAD molecules are fluorescent, changes in tissue auto-fluorescence provide valuable information on the cellular redox state and energy metabolism. Since fluorescence excitation, by mercury arc lamps (HBO) is inherently coupled to photo-bleaching and photo-toxicity, microfluorimetric monitoring of energy metabolism might benefit from the replacement of HBO lamps by light emitting diodes (LEDs). Here we describe a LED-based custom-built setup for monitoring NAD(P)H and FAD fluorescence at the level of single cells (HEK293) and of brain slices. We compared NAD(P)H bleaching characteristics with two light sources (HBO lamp and LED) as well as sensitivity and signal to noise ratio of three different detector types (multi-pixel photon counter (MPPC), photomultiplier tube (PMT) and photodiode). LED excitation resulted in reduced photo-bleaching at the same fluorescence output in comparison to excitation with the HBO lamp. Transiently increasing LED power resulted in reversible bleaching of NAD(P)H fluorescence. Recovery kinetics were dependent on metabolic substrates indicating coupling of NAD(P)H fluorescence to metabolism. Electrical stimulation of brain slices induced biphasic redox changes, as indicated by NAD(P)H/FAD fluorescence transients. Increasing the gain of PMT and decreasing the LED power resulted in similar sensitivity as obtained with the MPPC and the photodiode, without worsening the signal to noise ratio. In conclusion, replacement of HBO lamp with LED might improve conventional PMT based microfluorimetry of tissue auto-fluorescence.

  9. Fluorescence-based methods for detecting caries lesions: systematic review, meta-analysis and sources of heterogeneity.

    Directory of Open Access Journals (Sweden)

    Thais Gimenez

    Full Text Available BACKGROUND: Fluorescence-based methods have been proposed to aid caries lesion detection. Summarizing and analysing findings of studies about fluorescence-based methods could clarify their real benefits. OBJECTIVE: We aimed to perform a comprehensive systematic review and meta-analysis to evaluate the accuracy of fluorescence-based methods in detecting caries lesions. DATA SOURCE: Two independent reviewers searched PubMed, Embase and Scopus through June 2012 to identify papers/articles published. Other sources were checked to identify non-published literature. STUDY ELIGIBILITY CRITERIA, PARTICIPANTS AND DIAGNOSTIC METHODS: The eligibility criteria were studies that: (1 have assessed the accuracy of fluorescence-based methods of detecting caries lesions on occlusal, approximal or smooth surfaces, in both primary or permanent human teeth, in the laboratory or clinical setting; (2 have used a reference standard; and (3 have reported sufficient data relating to the sample size and the accuracy of methods. STUDY APPRAISAL AND SYNTHESIS METHODS: A diagnostic 2×2 table was extracted from included studies to calculate the pooled sensitivity, specificity and overall accuracy parameters (Diagnostic Odds Ratio and Summary Receiver-Operating curve. The analyses were performed separately for each method and different characteristics of the studies. The quality of the studies and heterogeneity were also evaluated. RESULTS: Seventy five studies met the inclusion criteria from the 434 articles initially identified. The search of the grey or non-published literature did not identify any further studies. In general, the analysis demonstrated that the fluorescence-based method tend to have similar accuracy for all types of teeth, dental surfaces or settings. There was a trend of better performance of fluorescence methods in detecting more advanced caries lesions. We also observed moderate to high heterogeneity and evidenced publication bias. CONCLUSIONS

  10. Analysis of simulated fluorescence intensities decays by a new maximum entropy method algorithm.

    Science.gov (United States)

    Esposito, Rosario; Altucci, Carlo; Velotta, Raffaele

    2013-01-01

    A new algorithm for the Maximum Entropy Method (MEM) is proposed for recovering the lifetime distribution in time-resolved fluorescence decays. The procedure is based on seeking the distribution that maximizes the Skilling entropy function subjected to the chi-squared constraint χ(2) ~ 1 through iterative linear approximations, LU decomposition of the Hessian matrix of the lagrangian problem and the Golden Section Search for backtracking. The accuracy of this algorithm has been investigated through comparisons with simulated fluorescence decays both of narrow and broad lifetime distributions. The proposed approach is capable to analyse datasets of up to 4,096 points with a discretization ranging from 100 to 1,000 lifetimes. A good agreement with non linear fitting estimates has been observed when the method has been applied to multi-exponential decays. Remarkable results have been also obtained for the broad lifetime distributions where the position is recovered with high accuracy and the distribution width is estimated within 3%. These results indicate that the procedure proposed generates MEM lifetime distributions that can be used to quantify the real heterogeneity of lifetimes in a sample.

  11. Stepwise multiphoton activation fluorescence reveals a new method of melanin detection

    Science.gov (United States)

    Lai, Zhenhua; Kerimo, Josef; Mega, Yair; DiMarzio, Charles A.

    2013-06-01

    The stepwise multiphoton activated fluorescence (SMPAF) of melanin, activated by a continuous-wave mode near infrared (NIR) laser, reveals a broad spectrum extending from the visible spectra to the NIR and has potential application for a low-cost, reliable method of detecting melanin. SMPAF images of melanin in mouse hair and skin are compared with conventional multiphoton fluorescence microscopy and confocal reflectance microscopy (CRM). By combining CRM with SMPAF, we can locate melanin reliably. However, we have the added benefit of eliminating background interference from other components inside mouse hair and skin. The melanin SMPAF signal from the mouse hair is a mixture of a two-photon process and a third-order process. The melanin SMPAF emission spectrum is activated by a 1505.9-nm laser light, and the resulting spectrum has a peak at 960 nm. The discovery of the emission peak may lead to a more energy-efficient method of background-free melanin detection with less photo-bleaching.

  12. Stepwise multiphoton activation fluorescence reveals a new method of melanin detection.

    Science.gov (United States)

    Lai, Zhenhua; Kerimo, Josef; Mega, Yair; Dimarzio, Charles A

    2013-06-01

    The stepwise multiphoton activated fluorescence (SMPAF) of melanin, activated by a continuous-wave mode near infrared (NIR) laser, reveals a broad spectrum extending from the visible spectra to the NIR and has potential application for a low-cost, reliable method of detecting melanin. SMPAF images of melanin in mouse hair and skin are compared with conventional multiphoton fluorescence microscopy and confocal reflectance microscopy (CRM). By combining CRM with SMPAF, we can locate melanin reliably. However, we have the added benefit of eliminating background interference from other components inside mouse hair and skin. The melanin SMPAF signal from the mouse hair is a mixture of a two-photon process and a third-order process. The melanin SMPAF emission spectrum is activated by a 1505.9-nm laser light, and the resulting spectrum has a peak at 960 nm. The discovery of the emission peak may lead to a more energy-efficient method of background-free melanin detection with less photo-bleaching.

  13. Design and daytime performance of laser-induced fluorescence spectrum lidar for simultaneous detection of multiple components, dissolved organic matter, phycocyanin, and chlorophyll in river water.

    Science.gov (United States)

    Saito, Yasunori; Kakuda, Kei; Yokoyama, Mizuho; Kubota, Tomoki; Tomida, Takayuki; Park, Ho-Dong

    2016-08-20

    In this work, we developed mobile laser-induced fluorescence spectrum (LIFS) lidar based on preliminary experiments on the excitation emission matrix of a water sample and a method for reducing solar background light using the synchronous detection technique. The combination of a UV short-pulse laser (355 nm, 6 ns) for fluorescence excitation with a 10-100 ns short-time synchronous detection using a gated image-intensified multi-channel CCD of the fluorescence made the LIFS lidar operation possible even in daytime. The LIFS lidar with this construction demonstrated the potential of natural river/lake water quality monitoring at the Tenryu River/Lake Suwa. Three main components in the fluorescence data of the water, dissolved organic matter, phycocyanin, and chlorophyll, were extracted by spectral analysis using the standard spectral functions of these components. Their concentrations were estimated by adapting experimentally calibrated data. Results of long-term field observations using our LIFS lidar from 2010 to 2012 show the necessity of simultaneous multi-component detection to understand the natural water environment.

  14. Synthesis and application of a N-1' fluorescent biotinyl derivative inducing the specific carboxy-terminal dual labeling of a novel RhoB-selective scFv.

    Science.gov (United States)

    Chaisemartin, L; Chinestra, P; Favre, G; Blonski, C; Faye, J C

    2009-05-20

    The fluorescent site-specific labeling of protein would provide a new, easy-to-use alternative to biochemical and immunochemical methods. We used an intein-mediated strategy for covalent labeling of the carboxy-terminal amino acid of a RhoB-selective scFv previously isolated from a phage display library (a human synthetic V(H) + V(L) scFv phage library). The scFv fused to the Mxe intein was produced in E. coli and purified and was then labeled with a newly synthesized fluorescent biotinyl cysteine derivative capable of inducing scFv-Mxe intein splicing. In this study, we investigated the splicing and labeling properties of various amino acids in the hinge domain between scFv and Mxe under thiol activation. In this dual labeling system, the fluorescein is used for antibody detection and biotin is used for purification, resulting in a high specific activity for fluorescence. We then checked that the purified biotinylated fluorescent scFv retained its selectivity for RhoB without modification of its affinity.

  15. Improved Savitzky-Golay-method-based fluorescence subtraction algorithm for rapid recovery of Raman spectra.

    Science.gov (United States)

    Chen, Kun; Zhang, Hongyuan; Wei, Haoyun; Li, Yan

    2014-08-20

    In this paper, we propose an improved subtraction algorithm for rapid recovery of Raman spectra that can substantially reduce the computation time. This algorithm is based on an improved Savitzky-Golay (SG) iterative smoothing method, which involves two key novel approaches: (a) the use of the Gauss-Seidel method and (b) the introduction of a relaxation factor into the iterative procedure. By applying a novel successive relaxation (SG-SR) iterative method to the relaxation factor, additional improvement in the convergence speed over the standard Savitzky-Golay procedure is realized. The proposed improved algorithm (the RIA-SG-SR algorithm), which uses SG-SR-based iteration instead of Savitzky-Golay iteration, has been optimized and validated with a mathematically simulated Raman spectrum, as well as experimentally measured Raman spectra from non-biological and biological samples. The method results in a significant reduction in computing cost while yielding consistent rejection of fluorescence and noise for spectra with low signal-to-fluorescence ratios and varied baselines. In the simulation, RIA-SG-SR achieved 1 order of magnitude improvement in iteration number and 2 orders of magnitude improvement in computation time compared with the range-independent background-subtraction algorithm (RIA). Furthermore the computation time of the experimentally measured raw Raman spectrum processing from skin tissue decreased from 6.72 to 0.094 s. In general, the processing of the SG-SR method can be conducted within dozens of milliseconds, which can provide a real-time procedure in practical situations.

  16. Validation of the chlorophyll fluorescence imaging method (CFI for early detection of herbicide resistance in weeds

    Directory of Open Access Journals (Sweden)

    Menegat, Alexander

    2014-02-01

    Full Text Available The increasing number of herbicide tolerant weed populations is illustrating the increasing demand for reliable methods for an accelerated detection of herbicide tolerance compared to greenhouse studies. Several methods for resistance quick detection have been published in previous years. One of the recent methods is the Chlorophyll Fluorescence Imaging Method (CFI. For this method changes in photosynthetic activity of the target organisms, caused by herbicides, are determined. General assumption of this method in terms of herbicide resistance detection is that each herbicidal compound, independent of the mode of action, will cause changes within the photosynthetic apparatus of the target organisms. This effect already could be confirmed for several modes of action (PSII, ALS, ACCase, EPSPS, synth. Auxins. Aim of this study is to validate this novel method on the basis of greenhouse experiments and single nucleotide polymorphisms (SNP analysis. The resistance profiles of 10 black-grass populations (Alopecurus myosuroides Huds. have been determined in greenhouse herbicide efficacy trials and constitutive SNP analyses of the survivors. With the CFI-method it was possible to detect the resistance profile as well as the resistance frequency within the populations. The results from the greenhouse experiments could be reproduced with conformity of 94%. This result is valid for the tested herbicides mesosulfuron, pyroxsulam as well as clodinafop and pinoxaden.

  17. Alignment-dependent fluorescence emission induced by tunnel ionization of carbon dioxide from lower-lying orbitals

    CERN Document Server

    Yao, Jinping; Jia, Xinyan; Hao, Xiaolei; Zeng, Bin; Jing, Chenrui; Chu, Wei; Ni, Jielei; Zhang, Haisu; Xie, Hongqiang; Zhang, Chaojin; Zhao, Zengxiu; Chen, Jing; Liu, Xiaojun; Cheng, Ya; Xu, Zhizhan

    2013-01-01

    We show that fluorescence emission induced by strong field tunnel ionization of carbon dioxide from its lower-lying orbitals exhibits a peculiar molecular alignment dependence. The experimentally measured alignment-dependence of the fluorescence agrees with the alignment-dependence of the ionization probability calculated in the framework of the strong field approximation. Our results demonstrate the feasibility of an all-optical approach for shedding more light on the ionization mechanisms of molecules from their lower-lying orbitals in tunnel ionization regime.

  18. Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Hui Su

    2001-05-01

    Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, we introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, we demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm{sub 2} for 40-{micro}m wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection.

  19. Laser-Induced Fluorescence Detection in High-Throughput Screening of Heterogeneous Catalysts and Single Cells Analysis

    Energy Technology Data Exchange (ETDEWEB)

    Su, Hui [Iowa State Univ., Ames, IA (United States)

    2001-01-01

    Laser-induced fluorescence detection is one of the most sensitive detection techniques and it has found enormous applications in various areas. The purpose of this research was to develop detection approaches based on laser-induced fluorescence detection in two different areas, heterogeneous catalysts screening and single cell study. First, we introduced laser-induced imaging (LIFI) as a high-throughput screening technique for heterogeneous catalysts to explore the use of this high-throughput screening technique in discovery and study of various heterogeneous catalyst systems. This scheme is based on the fact that the creation or the destruction of chemical bonds alters the fluorescence properties of suitably designed molecules. By irradiating the region immediately above the catalytic surface with a laser, the fluorescence intensity of a selected product or reactant can be imaged by a charge-coupled device (CCD) camera to follow the catalytic activity as a function of time and space. By screening the catalytic activity of vanadium pentoxide catalysts in oxidation of naphthalene, we demonstrated LIFI has good detection performance and the spatial and temporal resolution needed for high-throughput screening of heterogeneous catalysts. The sample packing density can reach up to 250 x 250 subunits/cm2 for 40-μm wells. This experimental set-up also can screen solid catalysts via near infrared thermography detection.

  20. A Proposed Method for Measurement of Absolute Air Fluorescence Yield based on High Resolution Optical Emission Spectroscopy

    CERN Document Server

    Gika, V; Maltezos, S

    2016-01-01

    In this work, we present a method for absolute measurement of air fluorescence yield based on high resolution optical emission spectroscopy. The absolute measurement of the air fluorescence yield is feasible using the Cherenkov light, emitted by an electron beam simultaneously with the fluorescence light, as a "standard candle". The separation of these two radiations can be accomplished exploiting the "dark" spectral regions of the emission band systems of the molecular spectrum of nitrogen. In these "dark" regions the net Cherenkov light can be recorded experimentally and be compared with the calculated one. The instrumentation for obtaining the nitrogen molecular spectra in high resolution and the noninvasive method for monitoring the rotational temperature of the emission process are also described. For the experimental evaluation of the molecular spectra analysis we used DC normal glow discharges in air performed in an appropriate spectral lamp considered as an air-fluorescence light emulator. The propose...

  1. [Fluorescence quenching method for the determination of p-hydroxyphenylpyruvic acid].

    Science.gov (United States)

    Wu, F; Fu, M; Wei, X; Yang, W; Hu, R; Guo, L

    2001-06-01

    A fluorescence quenching method for the determination of p-hydroxyphenylpyruvic acid with trytophan in the medium pH 11.01 NH3-NH4Cl is studied. The calibration curve is linear for p-hydroxyphenylpyruvic acid from 0 to 15 micrograms.mL-1 with lambda ex/lambda em = 285/356 nm. The detection limit is 0.37 microgram.mL-1. Twenty replicate determinations of solutions containing 10 micrograms.mL-1 p-hydroxyphenylpyruvic acid show a relative standard deviation of 1.2%. This method can be applied to the determination of p-hydroxyphenylpyruvic acid in serum with satisfactory results.

  2. Production of Superoxide in Bacteria Is Stress- and Cell State-Dependent: A Gating-Optimized Flow Cytometry Method that Minimizes ROS Measurement Artifacts with Fluorescent Dyes

    Science.gov (United States)

    McBee, Megan E.; Chionh, Yok H.; Sharaf, Mariam L.; Ho, Peiying; Cai, Maggie W. L.; Dedon, Peter C.

    2017-01-01

    The role of reactive oxygen species (ROS) in microbial metabolism and stress response has emerged as a major theme in microbiology and infectious disease. Reactive fluorescent dyes have the potential to advance the study of ROS in the complex intracellular environment, especially for high-content and high-throughput analyses. However, current dye-based approaches to measuring intracellular ROS have the potential for significant artifacts. Here, we describe a robust platform for flow cytometric quantification of ROS in bacteria using fluorescent dyes, with ROS measurements in 10s-of-1000s of individual cells under a variety of conditions. False positives and variability among sample types (e.g., bacterial species, stress conditions) are reduced with a flexible four-step gating scheme that accounts for side- and forward-scattered light (morphological changes), background fluorescence, DNA content, and dye uptake to identify cells producing ROS. Using CellROX Green dye with Escherichia coli, Mycobacterium smegmatis, and Mycobacterium bovis BCG as diverse model bacteria, we show that (1) the generation of a quantifiable CellROX Green signal for superoxide, but not hydrogen peroxide-induced hydroxyl radicals, validates this dye as a superoxide detector; (2) the level of dye-detectable superoxide does not correlate with cytotoxicity or antibiotic sensitivity; (3) the non-replicating, antibiotic tolerant state of nutrient-deprived mycobacteria is associated with high levels of superoxide; and (4) antibiotic-induced production of superoxide is idiosyncratic with regard to both the species and the physiological state of the bacteria. We also show that the gating method is applicable to other fluorescent indicator dyes, such as the 5-carboxyfluorescein diacetate acetoxymethyl ester and 5-cyano-2,3-ditolyl tetrazolium chloride for cellular esterase and reductive respiratory activities, respectively. These results demonstrate that properly controlled flow cytometry coupled

  3. Structure and intermolecular vibrations of 7-azaindole-water 2:1 complex in a supersonic jet expansion: Laser-induced fluorescence spectroscopy and quantum chemistry calculation

    Indian Academy of Sciences (India)

    Montu K Hazra; Moitrayee Mukherjee; V Ramanathan; Tapas Chakraborty

    2012-01-01

    Laser-induced fluorescence spectra of a 2:1 complex between 7-azaindole and water, known as `non-reactive dimer’ of the molecule, have been measured in a supersonic jet expansion. The dispersed fluorescence spectrum of the electronic origin band of the complex shows a very large number of low-frequency vibrational features corresponding to different intermolecular modes of the complex in the ground electronic state. Geometries of several possible isomeric structures of the complex and their vibrational frequencies at harmonic approximation were calculated by electronic structure theory method at MP2/6-31G∗∗ level. An excellent agreement is observed between the measured and calculated intermolecular vibrational mode frequencies for the energetically most favoured structure of the complex, where the water molecule is inserted within one of the two N$\\cdots$H-N hydrogen bonds of the 7AI dimer.

  4. [Application of PARAFAC method and 3-D fluorescence spectra in petroleum pollutant measurement and analysis].

    Science.gov (United States)

    Pan, Zhao; Wang, Yu-tian; Shao, Xiao-qing; Wu, Xi-jun; Yang, Li-li

    2012-03-01

    A method for identification and concentration measurement of petroleum pollutant by combining three-dimensional (3-D) fluorescence spectra with parallel factor analysis (PARAFAC) was proposed. The main emphasis of research was the measurement of coexisting different kinds of petroleum. The CCl4 solutions of a 0# diesel sample, a 97# gasoline sample, and a kerosene sample were used as measurement objects. The condition of multiple petroleum coexistence was simulated by petroleum solutions with different mixed ratios. The character of PARAFAC in complex mixture coexisting system analysis was studied. The spectra of three kinds of solutions and the spectra of gasoline-diesel mixed samples, diesel-kerosene mixed samples, and gas oline-diesel mixed with small counts of kerosene interference samples were analyzed respectively. The core consistency diagnostic method and residual sum of squares method were applied to calculate the number of factors in PARAFAC. In gasoline-diesel experiment, gasoline or diesel can be identified and measured as a whole respectively by 2-factors parallel factors analysis. In diesel-kerosene experiment, 2-factors parallel factors analysis can only obtain the characters of diesel, and the 3rd factor is needed to separate the kerosene spectral character from the mixture spectrum. When small counts of kerosene exist in gasoline-diesel solution, gasoline and diesel still can be identified and measured as principal components by a 2-factors parallel factor analysis, and the effect of interference on qualitative analysis is not significant. The experiment verified that the PARAFAC method can obtain characteristic spectrum of each kind of petroleum, and the concentration of petroleum in solutions can be predicted simultaneously, with recoveries shown in the paper. The results showed the possibility of petroleum pollutant identification and concentration measurement based on the 3-D fluorescence spectra and PARAFAC.

  5. The development of methods of analysis of documents on the basis of the methods of Raman spectroscopy and fluorescence analysis

    Science.gov (United States)

    Gorshkova, Kseniia O.; Tumkin, Ilya I.; Kirillova, Elizaveta O.; Panov, Maxim S.; Kochemirovsky, Vladimir A.

    2017-05-01

    The investigation of natural aging of writing inks printed on paper using Raman spectroscopy was performed. Based on the obtained dependencies of the Raman peak intensities ratios on the exposure time, the dye degradation model was proposed. It was suggested that there are several competing bond breaking and bond forming reactions corresponding to the characteristic vibration frequencies of the dye molecule that simultaneously occur during ink aging process. Also we propose a methodology based on the study of the optical properties of paper, particularly changes in the fluorescence of optical brighteners included in its composition as well as the paper reflectivity using spectrophotometric methods. These results can be implemented to develop the novel and promising method of criminology.

  6. Determination of intracellular levels of 6-mercaptopurine metabolites in erythrocytes utilizing capillary electrophoresis with laser-induced fluorescence detection.

    Science.gov (United States)

    Rabel, S R; Stobaugh, J F; Trueworthy, R

    1995-01-01

    Capillary electrophoresis proved to be a useful technique for the analysis of intracellular levels of 6-thioguanosine mono-, di-, and triphosphate with analysis times of 20 min. Conditions required for baseline separation of the thioguanine nucleotides consisted of a 25 mM KH2PO4 (pH 8.0) buffer and a separation voltage of +28 kV. Laser-induced fluorescence detection (lambda ex = 325 nm, lambda em = 410 nm) of the thioguanine nucleotide metabolites of 6-mercaptopurine (6-MP) was possible following oxidation of the thiol functionality. Tedious extraction procedures involving mercury cellulose resins or phenyl mercury adduct formation, which had been required previously for the selective extraction of thiopurines from erythrocytes, were unnecessary due to the overall specificity of the approach. However, the inclusion of 50 mM EDTA in the sample preparation was required to inhibit the anabolic/catabolic enzymatic activity, which was responsible for the degradation of the analytes. The method demonstrated linearity from 5 to 1700 pmol/100 microliters red blood cells for the three analytes (RSDs < or = 8%). The feasibility of the method was demonstrated for the quantitation of 6-thioguanine nucleotides in patients receiving either oral or intravenous 6-MP therapy.

  7. Usefulness of cyclodextrin media for the determination of alpha-cypermethrin by photochemically induced fluorescence: analytical applications to natural waters.

    Science.gov (United States)

    Mbaye, Moussa; Gaye Seye, Mame Diabou; Coly, Atanasse; Tine, Alphonse; Aaron, Jean-Jacques

    2009-06-01

    The photochemically induced fluorescence (PIF) spectral properties of alpha-cypermethrin in organic solvents (hexane, dichloromethane, acetonitrile, ethanol) and in cyclodextrin aqueous solutions (beta-CD and 2-hydroxypropyl-beta-CD, 2-HP-beta-CD) were investigated. The photolysis kinetics of alpha-cypermethrin were evaluated in the various media. The PIF signal was found to be significantly enhanced in the CD media relative to the organic solvents. The stoichiometry and the formation constants of the alpha-cypermethrin inclusion complexes formed with the CDs were determined. The analytical performances of the PIF method were improved in the presence of HP-beta-CD relative to the other media, and a CD-enhanced PIF analytical method was developed. The limits of detection and limits of quantification ranged, respectively, between 6 and 98 ng/mL and between 24 and 343 ng/mL, depending on the medium. Application to the analysis of tap water and Senegal natural water samples collected close to agricultural areas and spiked with alpha-cypermethrin yielded satisfactory recoveries going from about 77% to 98%. An interference study of foreign species, including pesticides and inorganic ions likely to be present in natural waters, was also carried out.

  8. Simultaneous Determination of Levodopa and Its Metabolite in Human Blood by Capillary Electrophoresis with Laser-induced Fluorescence Detection

    Institute of Scientific and Technical Information of China (English)

    ZHANG Ying-Xue; ZHANG Zhu-Jun; YANG Feng

    2008-01-01

    A rapid, sensitive and reproducible method is described for the analysis of levodopa and its metabolite dopamine (DA) in human blood. The influence of carbidopa as the inhibitor againist the decarboxylase activity on the metabolism has been also studied. After derivatization in a dark pulsator for 12 h at room temperature, the fluorescein isothiocyanate (FITC) derivative of levodopa and other components were separated by capillary zone electrophoresis (CZE) within 13 min and detected with laser-induced fluorescence (LIF). Under the optimum analysis conditions, the linear range is 3.0×10-8-4.0×10-6 mol/L and 1.0×10-8-2.0×10-6 mol/L for levodopa and DA,respectively. The detection limits of levodopa and DA were 7.8×10-9 mol/L (39.0 amol) and 3.1×10-9 mol/L(15.5 amol), respectively. The method was successfully applied to monitoring the levodopa and DA in human blood after one took tablets orally.

  9. Laser-induced fluorescence resonance energy transfer for analysis of the quality of a DNA double helix

    Science.gov (United States)

    Bregadze, V. G.; Melikishvili, Z. G.; Giorgadze, T. G.; Khutsishvili, I. G.; Khuskivadze, T. B.; Jaliashvili, Z. V.; Sigua, K. I.

    2016-11-01

    The goal of this work is to use the method of the laser-induced fluorescence resonance energy transfer (FRET) of electronic excitation in a donor-acceptor pair of intercalators, (acridine orange (AO) as a donor and ethidium bromide (EB) as an acceptor), for the quantitative analysis of the quality of a DNA double helix. This approach obtains a visual picture of the defects of the genetic apparatus of tissue cells, particularly those of skin cells in real time and it can be used for the diagnosis of skin diseases and also in cosmetology. Transition metal (TM) ions such as Cu(II), Cu(I), Ag(I), silver nanoparticles (AgNPs), photo- and thermo effects were used to cause double helix defects in DNA. The concentration of DNA sites after exposure to Cu(II), Cu(I), Ag(I) ions, AgNPs impact, as well as laser irradiation (λ  =  457 nm) and temperature, which are applicable for intercalation, were estimated in relative units. The nanoscale FRET method enables the estimation of the concentration of double helix areas with high stability, applicable for intercalation in DNA after it was subjected to stress effect. It provides the opportunity to compare DNA-s of (1) different origin; (2) with various degrees of damage; (3) being in various functional states.

  10. Application of HPLC combined with laser induced fluorescence for protein profile analysis of tissue homogenates in cervical cancer.

    Science.gov (United States)

    Bhat, Sujatha; Patil, Ajeetkumar; Rai, Lavanya; Kartha, V B; Chidangil, Santhosh

    2012-01-01

    A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF) technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA) to get statistical information on different tissue components. Discrimination of different stages of the samples was carried out by considering three parameters--scores of factor, spectral residual, and Mahalanobis Distance. Diagnostic accuracy of the method was evaluated using Receiver Operating Characteristic (ROC) analysis, and Youden's index (J) plots. The PCA results showed high sensitivity and specificity (~100) for cervical cancer diagnosis. ROC and Youden's index curves for both normal and malignant standard sets show good diagnostic accuracy with high AUC values. The statistical analysis has shown that the differences in protein profiles can be used to diagnose biochemical changes in the tissue, and thus can be readily applied for the detection of cervical cancer, even in situations where a histopathology examination is not easy because of nonavailability of experienced pathologists.

  11. Application of HPLC Combined with Laser Induced Fluorescence for Protein Profile Analysis of Tissue Homogenates in Cervical Cancer

    Directory of Open Access Journals (Sweden)

    Sujatha Bhat

    2012-01-01

    Full Text Available A highly objective method, High Performance Liquid Chromatography with Laser Induced Fluorescence (HPLC-LIF technique was used to study the protein profiles of normal and cervical cancer tissue homogenates. A total of 44 samples including normal cervical biopsy samples from the hysterectomy patients and the patients suffering from different stages of the cervical cancer were recorded by HPLC-LIF and analysed by Principle Component Analysis (PCA to get statistical information on different tissue components. Discrimination of different stages of the samples was carried out by considering three parameters—scores of factor, spectral residual, and Mahalanobis Distance. Diagnostic accuracy of the method was evaluated using Receiver Operating Characteristic (ROC analysis, and Youden's index (J plots. The PCA results showed high sensitivity and specificity (∼100 for cervical cancer diagnosis. ROC and Youden's index curves for both normal and malignant standard sets show good diagnostic accuracy with high AUC values. The statistical analysis has shown that the differences in protein profiles can be used to diagnose biochemical changes in the tissue, and thus can be readily applied for the detection of cervical cancer, even in situations where a histopathology examination is not easy because of nonavailability of experienced pathologists.

  12. Bioanalysis of captopril : Two sensitive high-performance liquid chromatographic methods with pre- or postcolumn fluorescent labeling

    NARCIS (Netherlands)

    Kok, R.J; Visser, Jan; Moolenaar, Frits; de Zeeuw, D; Meijer, D.K F

    1997-01-01

    This study describes the development and comparison of two HPLC methods for the analysis of the antihypertensive drug captopril. The first method is based on a precolumn derivatization of captopril with the fluorescent label monobromobimane (MBB). The second method is based on a postcolumn reaction

  13. Direct visualization of secretion from single bovine adrenal chromaffin cells by laser-induced native fluorescence imaging microscopy

    Energy Technology Data Exchange (ETDEWEB)

    Tong, W.; Yeung, E.S. [Ames Laboratory---USDOE and Department of Chemistry, Iowa State University, Ames, Iowa 50011 (United States)

    1998-03-01

    Direct visualization of the secretion process of individual bovine adrenal chromaffin cells was achieved with laser-induced native fluorescence imaging microscopy. By monitoring the native fluorescence of catecholamines excited by the 275 nm laser line with an intensified charge-coupled-device (CCD) camera, we obtained good temporal and spatial resolution simultaneously without using additional fluorescent probes. Large variations were found among individual cells in terms of the amounts of catecholamines secreted and the rates of secretion. Different regions of a cell also behave differently during the secretion process. However, the degree of this local heterogeneity is smaller than in neurons and neuralgia. The influence of deep-ultraviolet (UV) laser excitation on cells is also discussed. This quantitative imaging technique provides a useful noninvasive approach for the study of dynamic cellular changes and the understanding of the molecular mechanisms of secretory processes. {copyright} {ital 1998} {ital Society for Applied Spectroscopy}

  14. Correlative atomic force and confocal fluorescence microscopy: single molecule imaging and force induced spectral shifts (Conference Presentation)

    Science.gov (United States)

    Basché, Thomas; Hinze, Gerald; Stöttinger, Sven

    2016-09-01

    A grand challenge in nanoscience is to correlate structure or morphology of individual nano-sized objects with their photo-physical properties. An early example have been measurements of the emission spectra and polarization of single semiconductor quantum dots as well as their crystallographic structure by a combination of confocal fluorescence microscopy and transmission electron microscopy.[1] Recently, the simultaneous use of confocal fluorescence and atomic force microscopy (AFM) has allowed for correlating the morphology/conformation of individual nanoparticle oligomers or molecules with their photo-physics.[2, 3] In particular, we have employed the tip of an AFM cantilever to apply compressive stress to single molecules adsorbed on a surface and follow the effect of the impact on the electronic states of the molecule by fluorescence spectroscopy.[3] Quantum mechanical calculations corroborate that the spectral changes induced by the localized force can be associated to transitions among the different possible conformers of the adsorbed molecule.

  15. A Novel Real-time Fluorescence Mutant-allele-specific Amplification Method for Rapid Single Nucleotide Polymorphism Analysis

    Institute of Scientific and Technical Information of China (English)

    2006-01-01

    Current methods for single nucleotide polymorphism (SNP) analysis are timeconsuming and complicated. We aimed at development of one-step real-time fluorescence mutant-allele-specific amplification (MASA) method for rapid SNP analysis. The method is a marriage of two technologies: MASA primers for target DNA and a double-stranded DNA-selective fluorescent dye, SYBR Green I. Genotypes are separated according to the different threshold cycles of the wild-type and mutant primers. K-ras oncogene was used as a target to validate the feasibility of the method. The experimental results showed that the different genotypes can be clearly discriminated by the assay. The real-time fluorescence MASA method will have an enormous potential for fast and reliable SNP analysis due to its simplicity and low cost.

  16. The diagnostic capability of laser induced fluorescence in the characterization of excised breast tissues

    Science.gov (United States)

    Galmed, A. H.; Elshemey, Wael M.

    2017-08-01

    Differentiating between normal, benign and malignant excised breast tissues is one of the major worldwide challenges that need a quantitative, fast and reliable technique in order to avoid personal errors in diagnosis. Laser induced fluorescence (LIF) is a promising technique that has been applied for the characterization of biological tissues including breast tissue. Unfortunately, only few studies have adopted a quantitative approach that can be directly applied for breast tissue characterization. This work provides a quantitative means for such characterization via introduction of several LIF characterization parameters and determining the diagnostic accuracy of each parameter in the differentiation between normal, benign and malignant excised breast tissues. Extensive analysis on 41 lyophilized breast samples using scatter diagrams, cut-off values, diagnostic indices and receiver operating characteristic (ROC) curves, shows that some spectral parameters (peak height and area under the peak) are superior for characterization of normal, benign and malignant breast tissues with high sensitivity (up to 0.91), specificity (up to 0.91) and accuracy ranking (highly accurate).

  17. X-Ray Fluorescence and Laser-Induced Breakdown Spectroscopy analysis of Roman silver denarii

    Energy Technology Data Exchange (ETDEWEB)

    Pardini, L. [Istituto di Chimica dei Composti Organometallici del CNR, Research Area of Pisa, Via G. Moruzzi 1, 56124 Pisa (Italy); El Hassan, A. [National Institute for Laser- Enhanced Sciences (NILES), Cairo University Giza (Egypt); Ferretti, M. [Istituto per le Tecnologie Applicate ai Beni Culturali, Area della Ricerca del CNR di Montelibretti Roma (Italy); Foresta, A.; Legnaioli, S.; Lorenzetti, G. [Istituto di Chimica dei Composti Organometallici del CNR, Research Area of Pisa, Via G. Moruzzi 1, 56124 Pisa (Italy); Nebbia, E. [Universita degli Studi di Torino (Italy); Catalli, F. [Monetiere di Firenze, Museo Archeologico Nazionale Firenze (Italy); Harith, M.A. [National Institute for Laser- Enhanced Sciences (NILES), Cairo University Giza (Egypt); Diaz Pace, D. [Institute of Physics ' Arroyo Seco' , Faculty of Science, Tandil (Argentina); Anabitarte Garcia, F. [Photonics Engineering Group, University of Cantabria, Santander (Spain); Scuotto, M. [Dipartimento di Scienze Archeologiche, Via Galvani 1, 56126 Pisa (Italy); Palleschi, V., E-mail: vincenzo.palleschi@cnr.it [Istituto di Chimica dei Composti Organometallici del CNR, Research Area of Pisa, Via G. Moruzzi 1, 56124 Pisa (Italy); Dipartimento di Scienze Archeologiche, Via Galvani 1, 56126 Pisa (Italy)

    2012-08-15

    In this paper we present the results of a study performed on a large collection of silver Roman republican denarii, encompassing about two centuries of history. The joint use of Laser-Induced Breakdown Spectroscopy (LIBS) and X-Ray Fluorescence (XRF) spectroscopy allowed for an accurate determination of the coins' elemental composition; the measurements, performed mostly in situ at the 'Monetiere' in Florence, revealed a striking connection between the 'quality' of the silver alloy and some crucial contemporary events. This finding was used to classify a group of denarii whose dating was otherwise impossible. The comparison with other contemporary denarii disproves a recent theory on the origin of the so called 'serrated' denarii (denarii showing notched chisel marks on the edge of the coin). - Highlights: Black-Right-Pointing-Pointer We studied a large collection of Roman republican silver denarii. Black-Right-Pointing-Pointer XRF and LIBS allowed to determine the precious metal content of the coins. Black-Right-Pointing-Pointer A correlation of the 'quality' of the alloy with some contemporary events was found. Black-Right-Pointing-Pointer The study allowed to controvert a recent theory on the so called 'serrated' denarii.

  18. A two-photon laser induced fluorescence diagnostic with improved sensitivity, localization, and measurement rate

    Science.gov (United States)

    Elliott, Drew; Scime, Earl; Short, Zachary

    2016-10-01

    A two-photon absorption laser induced fluorescence diagnostic has been developed for measuring neutrals in fusion plasmas. Implementation of this diagnostic on the HIT-SI3 spheromak has demonstrated the sensitivity of the diagnostic and shown that measurements taken over several plasma pulses are possible. These measurements yielded an unexpected loss of signal when complex collection optics were utilized. Simulations show that this loss of signal can be explained by chromatic aberrations caused by the disparate Kr and D emission. This loss of signal has been addressed with the development of a new calibration scheme involving xenon gas. The Xe calibration scheme emission occurs at 656.00 nm while the deuterium emission is 656.09 nm. This nearly identical emission allows for advanced optical techniques such as confocal collection/injection and spatial filtering to be employed without loss of signal. Spatial filtering has been demonstrated to decrease noise while improving measurement localization, while confocal collection/injection allows for probing and measuring to occur through one viewport. The Xe scheme also allows for a Doppler-free hydrogen measurement. Doppler-free measurements eliminate the need to scan the laser spectrally thus greatly increasing the rate of measurement.

  19. Consolidated Laser-Induced Fluorescence Diagnostic Systems for the NASA Ames Arc Jet Facilities

    Science.gov (United States)

    Grinstead, Jay H.; Wilder, Michael C.; Porter, Barry J.; Brown, Jeffrey D.; Yeung, Dickson; Battazzo, Stephen J.; Brubaker, Timothy R.

    2016-01-01

    The spectroscopic diagnostic technique of two photon absorption laser-induced fluorescence (LIF) of atomic species for non-intrusive arc jet flow property measurement was first implemented at NASA Ames in the mid-1990s. In 2013-2014, NASA combined the agency's large-scale arc jet test capabilities at NASA Ames. Concurrent with that effort, the agency also sponsored a project to establish two comprehensive LIF diagnostic systems for the Aerodynamic Heating Facility (AHF) and Interaction Heating Facility (IHF) arc jets. The scope of the project enabled further engineering development of the existing IHF LIF system as well as the complete reconstruction of the AHF LIF system. The updated LIF systems are identical in design and capability. They represent the culmination of over 20 years of development experience in transitioning a specialized laboratory research tool into a measurement system for large-scale, high-demand test facilities. This paper will document the latest improvements of the LIF system design and demonstrations of the redeveloped AHF and IHF LIF systems.

  20. Measurement of fuel corrosion products using planar laser-induced fluorescence

    Science.gov (United States)

    Wantuck, Paul J.; Sappey, Andrew D.; Butt, Darryl P.

    1993-01-01

    Characterizing the corrosion behavior of nuclear fuel material in a high-temperature hydrogen environment is critical for ascertaining the operational performance of proposed nuclear thermal propulsion (NTP) concepts. In this paper, we describe an experimental study undertaken to develop and test non-intrusive, laser-based diagnostics for ultimately measuring the distribution of key gas-phase corrosion products expected to evolve during the exposure of NTP fuel to hydrogen. A laser ablation technique is used to produce high temperature, vapor plumes from uranium-free zirconium carbide (ZrC) and niobium carbide (NbC) forms for probing by various optical diagnostics including planar laser-induced fluorescence (PLIF). We discuss the laser ablation technique, results of plume emission measurements, and we describe both the actual and proposed planar LIF schemes for imaging constituents of the ablated ZrC and NbC plumes. Envisioned testing of the laser technique in rf-heated, high temperature gas streams is also discussed.

  1. Improving the monitoring of crop productivity using spaceborne solar-induced fluorescence.

    Science.gov (United States)

    Guan, Kaiyu; Berry, Joseph A; Zhang, Yongguang; Joiner, Joanna; Guanter, Luis; Badgley, Grayson; Lobell, David B

    2016-02-01

    Large-scale monitoring of crop growth and yield has important value for forecasting food production and prices and ensuring regional food security. A newly emerging satellite retrieval, solar-induced fluorescence (SIF) of chlorophyll, provides for the first time a direct measurement related to plant photosynthetic activity (i.e. electron transport rate). Here, we provide a framework to link SIF retrievals and crop yield, accounting for stoichiometry, photosynthetic pathways, and respiration losses. We apply this framework to estimate United States crop productivity for 2007-2012, where we use the spaceborne SIF retrievals from the Global Ozone Monitoring Experiment-2 satellite, benchmarked with county-level crop yield statistics, and compare it with various traditional crop monitoring approaches. We find that a SIF-based approach accounting for photosynthetic pathways (i.e. C3 and C4 crops) provides the best measure of crop productivity among these approaches, despite the fact that SIF sensors are not yet optimized for terrestrial applications. We further show that SIF provides the ability to infer the impacts of environmental stresses on autotrophic respiration and carbon-use-efficiency, with a substantial sensitivity of both to high temperatures. These results indicate new opportunities for improved mechanistic understanding of crop yield responses to climate variability and change. © 2015 John Wiley & Sons Ltd.

  2. Flow Property Measurement Using Laser-Induced Fluorescence in the NASA Ames Interaction Heating Facility

    Science.gov (United States)

    Grinstead, Jay Henderson; Porter, Barry J.; Carballo, Julio Enrique

    2011-01-01

    The spectroscopic diagnostic technique of two photon absorption laser-induced fluorescence (TALIF) of atomic species has been applied to single-point measurements of velocity and static temperature in the NASA Ames Interaction Heating Facility (IHF) arc jet. Excitation spectra of atomic oxygen and nitrogen were recorded while scanning a tunable dye laser over the absorption feature. Thirty excitation spectra were acquired during 8 arc jet runs at two facility operating conditions; the number of scans per run varied between 2 and 6. Curve fits to the spectra were analyzed to recover their Doppler shifts and widths, from which the flow velocities and static temperatures, respectively, were determined. An increase in the number of independent flow property pairs from each as-measured scan was obtained by extracting multiple lower-resolution scans. The larger population sample size enabled the mean property values and their uncertainties for each run to be characterized with greater confidence. The average plus or minus 2 sigma uncertainties in the mean velocities and temperatures for all 8 runs were plus or minus 1.4% and plus or minus 11%, respectively.

  3. Laser-induced fluorescence and FT-Raman spectroscopy for characterizing patinas on stone substrates.

    Science.gov (United States)

    Oujja, M; Vázquez-Calvo, C; Sanz, M; Álvarez de Buergo, M; Fort, R; Castillejo, M

    2012-02-01

    This article reports on a compositional investigation of stone patinas: thin colored layers applied for protective and/or aesthetic purposes on architectural or sculptural substrates of cultural heritage. The analysis and classification of patinas provide important information of historic and artistic interest, as their composition reflects local practices, the availabilities of different materials, and the development of technological knowledge during specific historical periods. Model patinas fabricated according to traditional procedures and applied onto limestone, and a historic patina sample from the main façade of the San Blas Monastery in Lerma (a village in the province of Burgos, Spain), were analyzed by laser-induced fluorescence and Fourier transform Raman spectroscopy. The results obtained demonstrate the ability of these two analytical techniques to identify the key components of each formulation and those of the reaction products which result from the chemical and mineralogical transformations that occur during aging, as well as to provide information that can aid the classification of different types of patinas.

  4. Wind-induced changes in the dynamics of fluorescent organic matter in the coastal NW Mediterranean.

    Science.gov (United States)

    Aparicio, Fran L; Nieto-Cid, Mar; Calvo, Eva; Pelejero, Carles; López-Sanz, Àngel; Pascual, Josep; Salat, Jordi; Sánchez-Pérez, Elvia D; La Fuente, Patricia De; Gasol, Josep M; Marrasé, Cèlia

    2017-12-31

    Marine biogeochemistry dynamics in coastal marine areas is strongly influenced by episodic events such as rain, intense winds, river discharges and anthropogenic activities. We evaluated in this study the importance of these forcing events on modulating seasonal changes in the marine biogeochemistry of the northwestern coast of the Mediterranean Sea, based on data gathered from a fixed coastal sampling station in the area. A 4-year (2011-2014) monthly sampling at four depths (0.5m, 20m, 50m and 80m) was performed to examine the time variability of several oceanographic variables: seawater temperature, salinity, inorganic nutrient concentrations (NO3(-), PO4(3-) and SiO2), chlorophyll a (Chl a), dissolved organic carbon (DOC) and fluorescent dissolved organic matter (FDOM). FDOM dynamics was predominantly influenced by upwelling events and mixing processes, driven by strong and characteristic wind episodes. SW wind episodes favored the upwelling of deeper and denser waters into the shallower shelf, providing a surplus of autochthonous humic-like material and inorganic nutrients, whereas northerlies favored the homogenization of the whole shelf water column by cooling and evaporation. These different wind-induced processes (deep water intrusion or mixing), reported along the four sampled years, determined a high interannual environmental variability in comparison with other Mediterranean sampling sites. Copyright © 2017 Elsevier B.V. All rights reserved.

  5. Fluorescent light induces neurodegeneration in the rodent nigrostriatal system but near infrared LED light does not.

    Science.gov (United States)

    Romeo, Stefania; Vitale, Flora; Viaggi, Cristina; di Marco, Stefano; Aloisi, Gabriella; Fasciani, Irene; Pardini, Carla; Pietrantoni, Ilaria; Di Paolo, Mattia; Riccitelli, Serena; Maccarone, Rita; Mattei, Claudia; Capannolo, Marta; Rossi, Mario; Capozzo, Annamaria; Corsini, Giovanni U; Scarnati, Eugenio; Lozzi, Luca; Vaglini, Francesca; Maggio, Roberto

    2017-05-01

    We investigated the effects of continuous artificial light exposure on the mouse substantia nigra (SN). A three month exposure of C57Bl/6J mice to white fluorescent light induced a 30% reduction in dopamine (DA) neurons in SN compared to controls, accompanied by a decrease of DA and its metabolites in the striatum. After six months of exposure, neurodegeneration progressed slightly, but the level of DA returned to the basal level, while the metabolites increased with respect to the control. Three month exposure to near infrared LED light (∼710nm) did not alter DA neurons in SN, nor did it decrease DA and its metabolites in the striatum. Furthermore mesencephalic cell viability, as tested by [(3)H]DA uptake, did not change. Finally, we observed that 710nm LED light, locally conveyed in the rat SN, could modulate the firing activity of extracellular-recorded DA neurons. These data suggest that light can be detrimental or beneficial to DA neurons in SN, depending on the source and wavelength. Copyright © 2017 Elsevier B.V. All rights reserved.

  6. Investigation of Gas Seeding for Planar Laser-Induced Fluorescence in Hypersonic Boundary Layers

    Science.gov (United States)

    Arisman, C. J.; Johansen, C. T.; Bathel, B. F.; Danehy, P. M.

    2015-01-01

    Numerical simulations of the gas-seeding strategies required for planar laser-induced fluorescence in a Mach 10 (approximately Mach 8.2 postshock) airflow were performed. The work was performed to understand and quantify the adverse effects associated with gas seeding and to assess various types of seed gas that could potentially be used in future experiments. In prior experiments, NO and NO2 were injected through a slot near the leading edge of a flatplate wedge model used in NASA Langley Research Center's 31 in. Mach 10 air tunnel facility. In this paper, nitric oxide, krypton, and iodine gases were simulated at various injection rates. Simulations showing the deflection of the velocity boundary layer for each of the cases are presented. Streamwise distributions of velocity and concentration boundary-layer thicknesses, as well as vertical distributions of velocity, temperature, and mass distributions, are presented for each of the cases. A comparison between simulated streamwise velocity profiles and experimentally obtained molecular tagging velocimetry profiles using a nitric oxide seeding strategy is performed to verify the influence of such a strategy on the boundary layer. The relative merits of the different seeding strategies are discussed. The results from a custom solver based on OpenFOAM version 2.2.1 are compared against results obtained from ANSYS® Fluent version 6.3.

  7. Consolidated Laser-Induced Fluorescence Diagnostic Systems for the NASA Ames Arc Jet Facilities

    Science.gov (United States)

    Grinstead, Jay; Wilder, Michael C.; Porter, Barry; Brown, Jeff; Yeung, Dickson; Battazzo, Steve; Brubaker, Tim

    2016-01-01

    The spectroscopic diagnostic technique of two photon absorption laser-induced fluorescence (TALIF) of atomic species for non-intrusive arc jet flow property measurement was first implemented at NASA Ames in the mid-1990s. Use of TALIF expanded at NASA Ames and to NASA Johnsons arc jet facility in the late 2000s. In 2013-2014, NASA combined the agency's large-scale arc jet test capabilities at NASA Ames. Concurrent with that effort, the agency also sponsored a project to establish two comprehensive LIF diagnostic systems for the Aerodynamic Heating Facility (AHF) and Interaction Heating Facility (IHF) arc jets. The scope of the project enabled further engineering development of the existing IHF LIF system as well as the complete reconstruction of the original AHF LIF system. The updated LIF systems are identical in design and capability. They represent the culmination of over 20 years of development experience in transitioning a specialized laboratory research tool into a measurement system for large-scale, high-demand test facilities. This paper documents the overall system design from measurement requirements to implementation. Representative data from the redeveloped AHF and IHF LIF systems are also presented.

  8. Multi-Site N-glycan mapping study 1: Capillary electrophoresis - laser induced fluorescence.

    Science.gov (United States)

    Szekrényes, Ákos; Park, SungAe Suhr; Santos, Marcia; Lew, Clarence; Jones, Aled; Haxo, Ted; Kimzey, Michael; Pourkaveh, Shiva; Szabó, Zoltán; Sosic, Zoran; Feng, Peng; Váradi, Csaba; de l'Escaille, François; Falmagne, Jean-Bernard; Sejwal, Preeti; Niedringhaus, Thomas; Michels, David; Freckleton, Gordon; Hamm, Melissa; Manuilov, Anastasiya; Schwartz, Melissa; Luo, Jiann-Kae; van Dyck, Jonathan; Leung, Pui-King; Olajos, Marcell; Gu, Yingmei; Gao, Kai; Wang, Wenbo; Wegstein, Jo; Tep, Samnang; Guttman, András

    2016-01-01

    An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra high-performance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established.

  9. Multi-Site N-glycan mapping study 1: Capillary electrophoresis – laser induced fluorescence

    Science.gov (United States)

    Szekrényes, Ákos; Park, SungAe Suhr; Santos, Marcia; Lew, Clarence; Jones, Aled; Haxo, Ted; Kimzey, Michael; Pourkaveh, Shiva; Szabó, Zoltán; Sosic, Zoran; Feng, Peng; Váradi, Csaba; de l'Escaille, François; Falmagne, Jean-Bernard; Sejwal, Preeti; Niedringhaus, Thomas; Michels, David; Freckleton, Gordon; Hamm, Melissa; Manuilov, Anastasiya; Schwartz, Melissa; Luo, Jiann-Kae; van Dyck, Jonathan; Leung, Pui-King; Olajos, Marcell; Gu, Yingmei; Gao, Kai; Wang, Wenbo; Wegstein, Jo; Tep, Samnang; Guttman, András

    2016-01-01

    An international team that included 20 independent laboratories from biopharmaceutical companies, universities, analytical contract laboratories and national authorities in the United States, Europe and Asia was formed to evaluate the reproducibility of sample preparation and analysis of N-glycans using capillary electrophoresis of 8-aminopyrene-1,3,6-trisulfonic acid (APTS)-labeled glycans with laser induced fluorescence (CE-LIF) detection (16 sites) and ultra high-performance liquid chromatography (UHPLC, 12 sites; results to be reported in a subsequent publication). All participants used the same lot of chemicals, samples, reagents, and columns/capillaries to run their assays. Migration time, peak area and peak area percent values were determined for all peaks with >0.1% peak area. Our results demonstrated low variability and high reproducibility, both, within any given site as well across all sites, which indicates that a standard N-glycan analysis platform appropriate for general use (clone selection, process development, lot release, etc.) within the industry can be established. PMID:26466659

  10. Real-time Fluorescence PCR Method for Detection of Burkholderia glumae from Rice

    Institute of Scientific and Technical Information of China (English)

    FANG Yuan; XU Li-hui; TIAN Wen-xiao; HUAI Yan; YU Shan-hong; LOU Miao-miao; XIE Guan-lin

    2009-01-01

    Burkholderia glumae causing seedling rot and grain rot of rice was listed as a plant quarantine disease of China in 2007. It's quite necessary to set up effective detection methods for the pathogen to manage further dispersal of this disease. The present study combined the real-time PCR method with classical PCR to increase the detecting efficiency, and to develop an accurate, rapid and sensitive method to detect the pathogen in the seed quarantine for effective management of the disease. The results showed that all the tested strains of B. glumae produced about 139 bp specific fragments by the real-time PCR and the general PCR methods, while others showed negative PCR result. The bacteria could be detected at the concentrations of 1×104 CFU/mL by general PCR method and at the concentrations below 100 CFU/mL by real-time fluorescence PCR method. B. glumae could be detected when the inoculated and healthy seeds were mixed with a proportion of 1:100.

  11. Confocal fluorescence microscopy investigation of visible emitting defects induced by electron beam lithography in LIF films

    Energy Technology Data Exchange (ETDEWEB)

    Montereali, R.M.; Bigotta, S.; Pace, A.; Piccinini, M. [ENEA, Divisione Fisica Applicata, Centro Ricerche Frascati, Frascati, RM (Italy); Burattini, E.; Grilli, A.; Raco, A. [Istituto Nazionale di Fisica Nucleare, Laboratori Nazionali di Fisica, Frascati, Rome (Italy); Giammatteo, M. [Unita' Istituto Nazionale di Fisica Nucleare, Frascati, RM (Italy)]|[L' Aquila Univ., L' Aquila (Italy). Centro di Microscopia Elettronica; Picozzi, P.; Santucci, S. [Unita' Istituto Nazionale di Fisica Nucleare, Frascati, RM (Italy)]|[L' Aquila Univ., L' Aquila (Italy). Dipt. di Fisica

    2000-07-01

    Low energy electron irradiation of lithium fluoride (LiF), in the form of bulk crystals and films, gives rise to the stable formation of primary F defects and aggregated color centers in a thin layer located at the surface of the investigated material. For the first time a confocal light scanning microscope (CLSM) in fluorescence mode was used to reconstruct the depth distribution of efficiently emitting laser active color centers in a stripe-like region induced by 12 and 16 keV electrons on LiF films thermally evaporated on glass. The formation of the F{sub 3}{sup +} and F{sub 2} aggregated defects appears restricted to the electron penetration and proportional to their energy depth profile, as obtained from Monte Carlo simulations. [Italian] L'irraggiamento con elettroni di bassa energia del fluoruro di litio (LiF), in forma di cristalli e film, induce la formazione di difetti primari F e centri di colore aggregati stabili in un sottile strato localizzato alla superficie del materiale investigato. Per la prima volta un microscopio confocale a scansione (CLSM) in modalita' fluorescenza e' stato usato per ricostruire la distribuzione di centri di colore laser attivi ad alta efficienza di emissione nel visibile, in strisce colorate ottenute con elettroni da 12 e 16 keV su film di LiF evaporati termicamente su vetro. La formazione dei difetti aggregati F2 e F3+ risulta ristretta spazialmente nella regione di penetrazione degli elettroni e proporzionale al profilo della distribuzione dell'energia da essi depositata, ricavata tramite simulazioni Monte Carlo.

  12. Production and laser-induced fluorescence spectroscopy (L.I.F.S.) of different Hypericum perforatum L. extracts

    Science.gov (United States)

    Skalkos, Dimitris; Filippidis, George; Kapsokalyvas, Dimitris; Meyer, Heiko; Papazoglou, Theodore; Karentzou, Eleni; Dimitriou, Heleni; Kalmanti, M.

    2005-04-01

    We are reporting elsewhere, the promising photodynamic effect of Hypericum perforatum L. extract (PMF) against T24, NBT-II tumor bladder cells, and HL-60 leukemic cells (using 630nm, and 530nm laser light respectively). The main advantages of the extract as a photosensitizer are its low cost, extensive availability, adequate solubility, minimal toxicity, and use with a range of wavelengths. Extraction of dry herb with methanol yields the methanolic extract (ME) in 11%, which is then fractionated using liquid / liquid extraction, yielding the polar methanolic fraction (PMF) in 9,9% overall yield. Hypericin, a photosensitizing ingredient of the herb, was found in these extracts in concentrations as low as 0,51%, and 0,57% respectively. Laser induced fluorescence spectra from the ME and PMF were recorded in order to evaluate their photodiagnostic capacity. An Argon-ion laser was employed for the excitation of the samples. It was shown that the extracts resulted in different fluorescence spectra related both to their intensity, and shape. The intensities of these spectra were only 8 times less compared to the fluorescence of pure hypericin. The dependence of the signal on the pH of the medium of pure hypericin and of PMF was also investigated in order to determine specific spectra variations. According to the results hypericin fluorescence signal fades smoothly in highly acidic medium, while it decreases sharply in highly basic environment. On the contrary PMF gives a slow decrease of fluorescence in both acidic and basic medium. These data suggest that PMF-induced fluorescence is highly sensitive in basic and acidic environment.

  13. OPTIMIZATION OF LAMBLIASIS MICROSCOPIC DIAGNOSTICS BY THE METHOD OF POLARIZED FLUORESCENCE FOR PATIENTS WITH ROSACEA AND URTICARIAL

    Directory of Open Access Journals (Sweden)

    Maryana Kovalchuk

    2013-07-01

    Full Text Available Introduction: There is little information about diagnosis of concurrent lambliasis in patients with rosacea and urticaria. We used method of polarized fluorescence to diagnose liambliasis, taking into account belonging of macromolecular structures of unicellular parasites Giardia lamblia to the optically active substances with the properties of liquid crystals. Material and Methods: Lambliasis was diagnosed on the basis of feces parasitological research and duodenal contents by methods of light and optic microscopy and polarized fluorescence in 105 patients with rosacea and urticaria. Research results were processed by the method of variation statistics in the Statgraf program by using Student’s criterion. Results: Search results of lamblia in patients with rosacea and urticaria depended on the conditions of its holding, patients’ preparation and from the previously received basic therapy if it consisted absorbents. Due to the fact that the fluorescence polarization as a physical method does not require the use of any generally toxic, dye- fluorochromes, qualitative cyto fluorescent analysis of lamblia in greeting microdrugs enables to distinguish vegetative forms of cysts. Conclussions: Polarized fluorescence method allows optimize the microscopic diagnosis of lambliasis, increasing its sensitivity. Previous preparation for the laboratory examination of Giardia lamblia is needed for the best exposure of vermin for patients with rosacea and urticaria.

  14. Photo-induced fluorescence of magnesium derivatives of tetracycline antibiotics in wastewater samples

    Energy Technology Data Exchange (ETDEWEB)

    Pena, A., E-mail: apena@ff.uc.pt [Group of Health Surveillance, Center of Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra (Portugal); Albert-Garcia, J.R. [Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia (Portugal); Silva, L.J.G.; Lino, C.M. [Group of Health Surveillance, Center of Pharmaceutical Studies, Faculty of Pharmacy, University of Coimbra (Portugal); Calatayud, J. Martinez [Department of Analytical Chemistry, Faculty of Chemistry, University of Valencia (Portugal)

    2010-07-15

    An analytical strategy, for the determination of tetracyclines (TCs), based on a HPLC system coupled with a photo-reactor followed by post-column derivatization was developed. Higher fluorescence emission after coupling the resulting photo-fragments with magnesium ions was observed for the determination of minocycline (MC), epitetracycline (ETC), tetracycline (TC) and doxycycline (DC). The manifold included a HPLC system with a photo-reactor (PTFE tubing helically coiled around a low-pressure mercury lamp), a mixing T-piece and a fluorescence detector. The derivatization reagent was delivered at 0.5 mL min{sup -1} by a pump. After HPLC separation using a gradient system with a mobile phase containing oxalic acid 0.02 M and acetonitrile, TCs were irradiated for 60 s, and the resulting photo-fragments were mixed with the post-column derivatization reagent, and the magnesium derivatives of TCs were detected by fluorimetry ({lambda}{sub exc} 386 nm, {lambda}{sub em} 500 nm). The results obtained showed a significant increase of sensitivity due to photodegration of TCs, 45.4%, 37.6% and 25.3% for MC, TC and ETC respectively. For DC an increase of only 1.5% was observed. The developed method was successfully applied to TCs determination in hospital and municipal wastewater samples using solid phase extraction with Oasis HLB cartridges. The LOQs were 0.25, 0.15, 01 and 0.25 {mu}g L{sup -1} for TC, ETC, MC and DC, respectively. The recovery values oscillated between 107.1% and 92.4% for fortification of 2.5 {mu}g L{sup -1} of each antibiotic.

  15. A novel pre-column fluorescent derivatization method for the sensitive determination of aristolochic acids in medicinal herbs by high-performance liquid chromatography with fluorescence detection.

    Science.gov (United States)

    Guo, Lin; Yue, Hao; Cai, Zongwei

    2010-09-21

    Aristolochic acids (AAs) are a family of structurally related nitrophenanthrene carboxylic acids existing in Aristolochia, Bragantia, and Asarum species. AAs have been proven to have nephrotoxic and carcinogenic toxicity. In this study, a novel pre-column fluorescence derivatization procedure followed by high-performance liquid chromatography-fluorescence detection (HPLC-FLD) is developed for the analysis of AAs in medicinal herbs. The nitro group in the phenanthrene ring of AAs was removed by NaBH(4) in water-THF (2:1, v/v), resulting in the corresponding aristolic acids. The analysis of AAs in medicinal herbs was based of the sensitive fluorescence detection of aristolic acids after the chemical derivatization. Because the produced aristolic acids are highly fluorescent the limit of detection (LOD) of AAI and AAII were lowered to 0.06 and 0.04 ng/mL, respectively, which is at least an order of magnitude lower than those in the reported HPLC and LC-MS methods. Good linearity with correlation coefficients higher than 0.997 were obtained for AAI and AII in the calibration ranges of 0.2-800 ng/mL. The derivatization conditions such as reaction temperature, time and the amount of NaBH(4) were optimized. The developed method provided satisfactory intra-day and inter-day precisions with RSDs less than 1.4% and 3.8%, respectively. The relative analytical error was less than 7% for the analysis of AAI and AAII in spiked matrix samples.

  16. Analysis of serotonin in brain microdialysates using capillary electrophoresis and native laser-induced fluorescence detection.

    Science.gov (United States)

    Benturquia, Nadia; Couderc, François; Sauvinet, Valérie; Orset, Cyrille; Parrot, Sandrine; Bayle, Christophe; Renaud, Bernard; Denoroy, Luc

    2005-03-01

    Serotonin or 5-hydroxytryptamine (5-HT) is a major neurotransmitter in the central nervous system. In this work, a method for analyzing 5-HT in brain microdialysis samples using a commercially available capillary electrophoresis (CE) system has been developed. A pH-mediated in-capillary preconcentration of samples was performed, and after separation by capillary zone electrophoresis, native fluorescence of 5-HT was detected by a 266 nm solid-state laser. The separation conditions for the analysis of 5-HT in standard solutions and microdialysates have been optimized, and this method has been validated on both pharmacological and analytical bases. Separation of 5-HT was performed using a 80 mmol/L citrate buffer, pH 2.5, containing 20 mmol/L hydroxypropyl-beta-cyclodextrin (HP-beta-CD) and +30 kV voltage. The detection limit was 2.5 x 10(-10) mol/L. This method allows the in vivo brain monitoring of 5-HT using a simple, accurate CE measurement in underivatized microdialysis samples.

  17. Time-resolved detection of aromatic compounds on planetary surfaces by ultraviolet laser induced fluorescence and Raman spectroscopy

    Science.gov (United States)

    Eshelman, E.; Daly, M. G.; Slater, G.; Cloutis, E.

    2015-12-01

    Raman spectroscopic instruments are highly capable in the search for organics on Mars due to the potential to perform rapid and nondestructive measurements on unprepared samples. Upcoming and future Raman instruments are likely to also incorporate laser-induced fluorescence (LIF) capabilities, which can be added for modest cost and complexity. We demonstrate that it is possible to obtain sub-ns fluorescence lifetime measurements of Mars-relevant organics and minerals if a fast time-gating capability is used with an intensified detector and a short ultraviolet laser pulse. This serves a primary purpose of discriminating mineral from short-lived (less than 10 ns) organic fluorescence, considered a potential biosignature. Additionally, lifetime measurements may assist in determining if more than one fluorescing species is present and provide information concerning the molecular structure as well as the local environment. Fast time-gating is also useful at longer visible or near-IR wavelengths, as this approach increases the sensitivity of the instrument to organic material by removing the majority of the fluorescence background from the Raman signal and reducing the effect of ambient light.

  18. Ag@Aggregation-induced emission dye core/shell nanostructures with enhanced one- and two-photon fluorescence

    Science.gov (United States)

    Wang, Cheng; Li, Yang; Xu, Qiujin; Luo, Liang

    2017-10-01

    Combining plasmonic nanostructures with two-photon fluorescence materials is a promising way to significantly enhance two-photon fluorescence. Ag@1,4-bis(2-cyano-2-phenylethenyl) benzene (BCPEB) core/shell nanostructures were fabricated by simply incubating the isolated Ag nanoparticles with BCPEB microrods in ethanol. BCPEB was chosen as the fluorescent organic molecule owing to the aggregation-induced-emission (AIE) nature which would reduce the emission loss as being practically applied in solid phase. By utilizing the match of the extinction spectrum of Ag nanoparticles and BCPEB's absorption band, the target Ag@BCPEB core/shell nanostructures showed an enhanced one-photon (12×) fluorescence, integrating with SERS signal as well. Moreover, the resultant second harmonic generation of Ag nanoparticles under two-photon excitation also well matched with the absorption band of BCPEB, and significant enhanced two-photon (17×) fluorescence was obtained. The confocal images of NIH-3T3 cells with these nanostructures under one- and two-photon excitation showed good contrast and brightness for bio-imaging.

  19. Spectrum reconstruction method based on the detector response model calibrated by x-ray fluorescence

    Science.gov (United States)

    Li, Ruizhe; Li, Liang; Chen, Zhiqiang

    2017-02-01

    Accurate estimation of distortion-free spectra is important but difficult in various applications, especially for spectral computed tomography. Two key problems must be solved to reconstruct the incident spectrum. One is the acquisition of the detector energy response. It can be calculated by Monte Carlo simulation, which requires detailed modeling of the detector system and a high computational power. It can also be acquired by establishing a parametric response model and be calibrated using monochromatic x-ray sources, such as synchrotron sources or radioactive isotopes. However, these monochromatic sources are difficult to obtain. Inspired by x-ray fluorescence (XRF) spectrum modeling, we propose a feasible method to obtain the detector energy response based on an optimized parametric model for CdZnTe or CdTe detectors. The other key problem is the reconstruction of the incident spectrum with the detector response. Directly obtaining an accurate solution from noisy data is difficult because the reconstruction problem is severely ill-posed. Different from the existing spectrum stripping method, a maximum likelihood-expectation maximization iterative algorithm is developed based on the Poisson noise model of the system. Simulation and experiment results show that our method is effective for spectrum reconstruction and markedly increases the accuracy of XRF spectra compared with the spectrum stripping method. The applicability of the proposed method is discussed, and promising results are presented.

  20. Two Photon Absorption Laser Induced Fluorescence for Neutral Hydrogen Profile Measurements

    Energy Technology Data Exchange (ETDEWEB)

    Scime, Earl E. [West Virginia Univ., Morgantown, WV (United States)

    2016-09-23

    The magnitude and spatial dependence of neutral density in magnetic confinement fusion experiments is a key physical parameter, particularly in the plasma edge. Modeling codes require precise measurements of the neutral density to calculate charge-exchange power losses and drag forces on rotating plasmas. However, direct measurements of the neutral density are problematic. In this work, we proposed to construct a laser-based diagnostic capable of providing spatially resolved measurements of the neutral density in the edge of plasma in the DIII-D tokamak. The diagnostic concept is based on two-photon absorption laser induced fluorescence (TALIF). By injecting two beams of 205 nm light (co or counter propagating), ground state hydrogen (or deuterium or tritium) can be excited from the n = 1 level to the n = 3 level at the location where the two beams intersect. Individually, the beams experience no absorption, and therefore have no difficulty penetrating even dense plasmas. After excitation, a fraction of the hydrogen atoms decay from the n = 3 level to the n = 2 level and emit photons at 656 nm (the Hα line). Calculations based on the results of previous TALIF experiments in magnetic fusion devices indicated that a laser pulse energy of approximately 3 mJ delivered in 5 ns would provide sufficient signal-to-noise for detection of the fluorescence. In collaboration with the DIII-D engineering staff and experts in plasma edge diagnostics for DIII-D from Oak Ridge National Laboratory (ORNL), WVU researchers designed a TALIF system capable of providing spatially resolved measurements of neutral deuterium densities in the DIII-D edge plasma. The laser systems were specified, purchased, and assembled at WVU. The TALIF system was tested on a low-power hydrogen discharge at WVU and the plan was to move the instrument to DIII-D for installation in collaboration with ORNL researchers. After budget cuts at DIII-D, the DIII-D facility declined to support

  1. Detection and characterization of chemical-induced abnormal tissue and rat tumors at different stages using fluorescence spectroscopy

    Science.gov (United States)

    Chen, Wei R.; Jassemnejad, Baha; Crull, Jason; Knobbe, Edward T.; Nordquist, Robert E.

    1996-04-01

    Fluorescence spectroscopy of diseased tissues, including chemical-induced rat liver, kidney and testis lesions, as well as murine mammary tumor, was studied. The rat liver, kidney and testis tissues were excited by radiation of 350 and 366 nm, which appeared to provide the optimal differentiation between normal and lesion tissues; the tumor tissues were excited by both 350 nm and 775 nm wavelengths. In comparison with normal liver tissue, at (lambda) ex equals 366 nm, the fluorescent spectrum of liver lesion showed a clear red shift around the emission peak of 470 nm, the major native fluorescent peak of organized tissue. When excited by 350 nm wavelength, all the chemically induced lesion tissues (liver, kidney and testis) appeared to cause a significant reduction of emission intensity at the 470 nm peak. While the 775 nm excitation did not reveal any significant difference among tumor, muscle and skin tissues, the 350 nm excitation did provide some interesting features among the tumor tissues at different stages. Compared with muscle tissue, the viable tumor showed an overall reduction of emission intensity around 470 nm. In addition, the viable tumor tissue showed a secondary emission peak at 390 nm with necrotic tumor tissue having a reduced intensity. The histology of both viable and necrotic tumor tissue was examined and appeared to correlate with the results of the fluorescent spectroscopy observation.

  2. Rapid and Inexpensive Screening of Genomic Copy Number Variations Using a Novel Quantitative Fluorescent PCR Method

    Directory of Open Access Journals (Sweden)

    Martin Stofanko

    2013-01-01

    Full Text Available Detection of human microdeletion and microduplication syndromes poses significant burden on public healthcare systems in developing countries. With genome-wide diagnostic assays frequently inaccessible, targeted low-cost PCR-based approaches are preferred. However, their reproducibility depends on equally efficient amplification using a number of target and control primers. To address this, the recently described technique called Microdeletion/Microduplication Quantitative Fluorescent PCR (MQF-PCR was shown to reliably detect four human syndromes by quantifying DNA amplification in an internally controlled PCR reaction. Here, we confirm its utility in the detection of eight human microdeletion syndromes, including the more common WAGR, Smith-Magenis, and Potocki-Lupski syndromes with 100% sensitivity and 100% specificity. We present selection, design, and performance evaluation of detection primers using variety of approaches. We conclude that MQF-PCR is an easily adaptable method for detection of human pathological chromosomal aberrations.

  3. Improved method to raise polyclonal antibody using enhanced green fluorescent protein transgenic mice

    Institute of Scientific and Technical Information of China (English)

    Jianke Ren; Long Wang; Guoxiang Liu; Wen Zhang; Zhejin Sheng; Zhugang Wang; Jian Fei

    2008-01-01

    Recombinant fusion protein is widely used as an antigen to raise antibodies against the epitope of a target protein. However, the concomitant anticarrier antibody in resulting antiserum reduces the production of the desired antibody and brings about unwanted non-specific immune reactions. It is proposed that the carrier protein transgenic animal could be used to solve this problem. To validate this hypothesis, enhanced green fluorescent protein (EGFP) transgenic mice were produced. By immunizing the mice with fusion protein His6HAtag-EGFP, we showed that the antiserum from the transgenic mice had higher titer antibody against His6HA tag and lower titer antibody against EGFP compared with that from wild-type mice. Therefore, this report describes an improved method to raise high titer antipeptide polyclonal antibody using EGFP transgenic mice that could have application potential in antibodypreparation.

  4. Study of Organic Matter in Soils of the Amazon Region Employing Laser Induced Fluorescence Spectroscopy

    Science.gov (United States)

    Tadini, Amanda Maria; Nicolodelli, Gustavo; Mounier, Stéphane; Montes, Célia Regina; Marcondes Bastos Pereira Milori, Débora

    2014-05-01

    In the face of climate change and increasing CO2 levels in the atmosphere, the global carbon cycle, soil organic carbon (SOC) sequestration, and the role of different world biomes as potential sources and sinks of carbon are receiving increasing attention. Carbon quantification is an important environmental indicator, but the structure of organic matter is also important because is related to carbon stability. The synthesis of soil organic matter (SOM), as presented in soils of forest vegetation, can be originated from condensation polymeric polyphenols and quinones that are responsible for controlling the main physical-chemical properties of soils. These systems are present in humic substances, representing the major fluorophore of SOM[1-3]. Abiotic factors, such as soil texture, use and occupation of soil, can influence on the process of SOM formation, molecular structure and in its humification index[4]. Laser Induced Fluorescence Spectroscopy (LIFS) have become a promising technique for assessing humification index of SOM (HLIFS). In this context, the aim of this study was to analyze the humification index of the SOM in the region of Barcelos (Amazon) employing LIFS. The study area was the region of Barcelos, close the river Demeni. The whose vegetation distribution in this area, is two biomes the Dense Ombrophylous Forest (DPQD) and Campinarana (DPQT), with areas of edaphic contacts between these two phytophysiognomies, which ranged from Open field (FDE) to closed Depression (DPQ). Preliminary results showed that the area closed Depression (DPQ) there was a continuous gradient of humification with increasing soil depth. A similar behavior was verified for area Forest (DPQD), where the highest values of HLIFS were obtained between the four points analyzed, indicating the magnitude of the molecular recalcitrance this organic matter in this area. The results obtained for area Campinarana (DPQT) and Open field (FDE) showed an opposite behavior. These points there

  5. Towards a Novel Spatially-Resolved Hemolysis Detection Method Using a Fluorescent Indicator and Loaded Ghost Cells: Proof-of-Principle.

    Science.gov (United States)

    Jansen, Sebastian V; Müller, Indra; Kiesendahl, Nicole; Schmitz-Rode, Thomas; Steinseifer, Ulrich

    2015-09-01

    It is of the utmost importance to reduce flow-induced hemolysis in devices such as heart-valve prostheses and blood pumps. Thus, in vitro measurements of hemolysis are performed in order to optimize their design in this regard. However, with existing measurement methods, hemolysis can only be assessed as an integrated value over the complete test-circuit. Currently, there are no spatially-resolved in vitro hemolysis measurement techniques known to the authors that would allow for a determination of the critical regions within a device. In this study, a novel spatially-resolved measurement principle is proposed. Ghost cells (i.e. erythrocytes with a lower hemoglobin concentration) were loaded with a calcium-dicitrato complex, and a fluorescent calcium indicator was suspended in the extracellular medium. Calcium and indicator are separated until the cell membrane ruptures (i.e. hemolysis occurs). In the moment of hemolysis, the two compounds bind to each other and emit a fluorescent signal that can be recorded and spatially-resolved in a setup very similar to a standard Particle Image Velocimetry measurement. A proof-of-principle experiment was performed by intentionally inducing hemolysis in a flow-model with a surfactant. The surfactant-induced hemolysis demonstrated a clear increase of the fluorescent signal compared to that of a negative reference. Furthermore, the signal was spatially restricted to the area of hemolysis. Although further challenges need to be addressed, a successful proof-of-principle for novel spatially-resolved hemolysis detection is presented. This method can contribute to better design optimization of devices with respect to flow-induced hemolysis.

  6. Fluorescence studies of ligand-induced conformational changes of the Na(+)/glucose cotransporter

    DEFF Research Database (Denmark)

    Meinild, Anne-Kristine; Hirayama, Bruce A; Wright, Ernest M;

    2002-01-01

    (+) also increased fluorescence, whereas choline, tetraethylammonium, and N-methyl-D-glucamine did not. Fluorescence was increased by sugars with specificity: methyl alpha-D-glucopyranoside > D-glucose > D-galactose >> D-mannitol. Voltage-jump experiments (in 100 mM NaCl buffer in absence of sugar...

  7. Investigation of Laser Induced Fluorescence for Concentration Measurements of Diatomic Sulfur.

    Science.gov (United States)

    1981-12-01

    Once the spectrum was obtained, it was calibrated with N2 laser scatter and a mercury reference run. Fluorescence peaks were identified by com... Freddie , Jr. The Measurement of Quenching Rate Constants Using Fluorescence Emission. MS Thesis. Wright-Patterson Air Force Base, Ohio: Air Force Institute

  8. Studies of the laser-induced fluorescence of explosives and explosive compositions.

    Energy Technology Data Exchange (ETDEWEB)

    Hargis, Philip Joseph, Jr. (,; .); Thorne, Lawrence R.; Phifer, Carol Celeste; Parmeter, John Ethan; Schmitt, Randal L.

    2006-10-01

    Continuing use of explosives by terrorists throughout the world has led to great interest in explosives detection technology, especially in technologies that have potential for standoff detection. This LDRD was undertaken in order to investigate the possible detection of explosive particulates at safe standoff distances in an attempt to identify vehicles that might contain large vehicle bombs (LVBs). The explosives investigated have included the common homogeneous or molecular explosives, 2,4,6-trinitrotoluene (TNT), pentaerythritol tetranitrate (PETN), cyclonite or hexogen (RDX), octogen (HMX), and the heterogeneous explosive, ammonium nitrate/fuel oil (ANFO), and its components. We have investigated standard excited/dispersed fluorescence, laser-excited prompt and delayed dispersed fluorescence using excitation wavelengths of 266 and 355 nm, the effects of polarization of the laser excitation light, and fluorescence imaging microscopy using 365- and 470-nm excitation. The four nitro-based, homogeneous explosives (TNT, PETN, RDX, and HMX) exhibit virtually no native fluorescence, but do exhibit quenching effects of varying magnitude when adsorbed on fluorescing surfaces. Ammonium nitrate and fuel oil mixtures fluoresce primarily due to the fuel oil, and, in some cases, due to the presence of hydrophobic coatings on ammonium nitrate prill or impurities in the ammonium nitrate itself. Pure ammonium nitrate shows no detectable fluorescence. These results are of scientific interest, but they provide little hope for the use of UV-excited fluorescence as a technique to perform safe standoff detection of adsorbed explosive particulates under real-world conditions with a useful degree of reliability.

  9. Studies on the Transition Metal Ion Induced Fluorescence Enhancement of 1,8-Naphthalimide Derivatives

    Institute of Scientific and Technical Information of China (English)

    WEN,Guo-Tao; ZHU,Man-Zhou; WANG,Zhuo; MENG,Xiang-Ming; HU,Hui-Yuan; GUO,Qing-Xiang

    2006-01-01

    Two fluorosensor systems have been designed and synthesized with their photophysical properties and fluorescence responses toward the transition metal ions studied. The fluorosensor was composed of 1,8-naphthalimide and 4-chloro-1,8-naphthalimide as fluorophore respectively, an amino moiety as the receptor and a hydrocarbon chain as the spacer to link the fluorophore and receptor. Fluorescence intensity of these systems is very weak due to the process of the efficient intramolecular photoinduced electron transfer (PET) in the absence of metal ions. Titration of the transition metal ions can switch on the fluorescence immediately. The intensity of released fluorescence is even higher than the expected from the consideration of the PET in these systems. It may be rationalized that the receptor bound to transition metal ions and the solvation of the fluorophore by the water molecules from the hydrated transition metal salts, may significantly cause fluorescence enhancement.

  10. Visualization of Structured Packing with Laser Induced Fluorescence Technique:Two-Dimensional Measurement of Liquid Concentration Distribution

    Institute of Scientific and Technical Information of China (English)

    刘伯潭; 申言同; 张会书; 刘春江; 唐忠利; 袁希钢

    2016-01-01

    A method of using laser induced fluorescence(LIF)technique was applied to two-dimensional meas-urement of the liquid concentration distribution in the 250Y structured packing sheet. The experimental structured packing sheet was made of perspex so that the laser could pass through it. The visualization of the distribution of the liquid concentration in the structured packing sheet was realized. The calibration of the thickness and liquid concentration was carried out firstly and the regression formulaI=kcd was acquired, in which concentrationc and the liquid film thicknessd were both considered. Then the liquid feed of uniform tracer(rhodamine)concentration entered the perspex structured packing from the top under different spraying densities. The corresponding thickness of liquid film on the packing was calculated. Finally, tracer(rhodamine)with a high concentration was injected only at one fixed point of the structured packing under different spraying densities of the liquid. With the known liquid film thickness, the concentration distribution of the tracer can be calculated inside the structured packing sheet.

  11. Non-rigid contour-to-pixel registration of photographic and quantitative light-induced fluorescence imaging of decalcified teeth

    Science.gov (United States)

    Berkels, Benjamin; Deserno, Thomas; Ehrlich, Eva E.; Fritz, Ulrike B.; Sirazitdinova, Ekaterina; Tatano, Rosalia

    2016-03-01

    Quantitative light-induced fluorescence (QLF) is widely used to assess the damage of a tooth due to decalcification. In digital photographs, decalcification appears as white spot lesions, i.e. white spots on the tooth surface. We propose a novel multimodal registration approach for the matching of digital photographs and QLF images of decalcified teeth. The registration is based on the idea of contour-to-pixel matching. Here, the curve, which represents the shape of the tooth, is extracted from the QLF image using a contour segmentation by binarization and morphological processing. This curve is aligned to the photo with a non-rigid variational registration approach. Thus, the registration problem is formulated as minimization problem with an objective function that consists of a data term and a regularizer for the deformation. To construct the data term, the photo is pointwise classified into tooth and non-tooth regions. Then, the signed distance function of the tooth region allows to measure the mismatch between curve and photo. As regularizer a higher order, linear elastic prior is used. The resulting minimization problem is solved numerically using bilinear Finite Elements for the spatial discretization and the Gauss-Newton algorithm. The evaluation is based on 150 image pairs, where an average of 5 teeth have been captured from 32 subjects. All registrations have been confirmed correctly by a dental expert. The contour-to-pixel methods can directly be used in 3D for surface-to-voxel tasks.

  12. Temporal Unmixing of Dynamic Fluorescent Images by Blind Source Separation Method with a Convex Framework

    Directory of Open Access Journals (Sweden)

    Duofang Chen

    2015-01-01

    Full Text Available By recording a time series of tomographic images, dynamic fluorescence molecular tomography (FMT allows exploring perfusion, biodistribution, and pharmacokinetics of labeled substances in vivo. Usually, dynamic tomographic images are first reconstructed frame by frame, and then unmixing based on principle component analysis (PCA or independent component analysis (ICA is performed to detect and visualize functional structures with different kinetic patterns. PCA and ICA assume sources are statistically uncorrelated or independent and don’t perform well when correlated sources are present. In this paper, we deduce the relationship between the measured imaging data and the kinetic patterns and present a temporal unmixing approach, which is based on nonnegative blind source separation (BSS method with a convex analysis framework to separate the measured data. The presented method requires no assumption on source independence or zero correlations. Several numerical simulations and phantom experiments are conducted to investigate the performance of the proposed temporal unmixing method. The results indicate that it is feasible to unmix the measured data before the tomographic reconstruction and the BSS based method provides better unmixing quality compared with PCA and ICA.

  13. Diagnosis of bacterial vaginosis by a new multiplex peptide nucleic acid fluorescence in situ hybridization method

    Science.gov (United States)

    Machado, António; Castro, Joana; Cereija, Tatiana; Almeida, Carina

    2015-01-01

    Bacterial vaginosis (BV) is one of most common vaginal infections. However, its diagnosis by classical methods reveals low specificity. Our goal was to evaluate the accuracy diagnosis of 150 vaginal samples with research gold standard methods and our Peptide Nucleic Acid (PNA) probes by Fluorescence in situ Hybridization (FISH) methodology. Also, we described the first PNA-FISH methodology for BV diagnosis, which provides results in approximately 3 h. The results showed a sensitivity of 84.6% (95% confidence interval (CI), from 64.3 to 95.0%) and a specificity of 97.6% (95% CI [92.6–99.4%]), demonstrating the higher specificity of the PNA-FISH method and showing false positive results in BV diagnosis commonly obtained by the classical methods. This methodology combines the specificity of PNA probes for Lactobacillus species and G. vaginalis visualization and the calculation of the microscopic field by Nugent score, allowing a trustful evaluation of the bacteria present in vaginal microflora and avoiding the occurrence of misleading diagnostics. Therefore, the PNA-FISH methodology represents a valuable alternative for BV diagnosis. PMID:25737820

  14. New Fluorescent Nanoparticles for Ultrasensitive Detection of Nucleic Acids by Optical Methods

    DEFF Research Database (Denmark)

    Westergaard Mulberg, Mads; Taskova, Maria; Thomsen, Rasmus P.

    2017-01-01

    in the synthesis of new fluorescent nanoparticles. Here, the fluorescent nanoparticles were made by simple and inexpensive radical emulsion polymerization of butyl acrylate in the presence of fluorescent dyes and additional functionalization reagents. This provided ultra-bright macrofluorophores of 9-84nm mean...... diameter, modified with additional alkyne and amino groups for bioconjugation. By using click and NHS chemistries, the new nanoparticles were attached to target-specific DNA probes that were used in fluorimetry and fluorescence microscopy. Overall, these fluorescent nanoparticles and their oligonucleotide...... derivatives have higher photostability, brighter fluorescence and hence dramatically lower limits of target detection than the individual organic dyes. These properties make them useful in approaches directed towards ultrasensitive detection of nucleic acids, in particular for imaging and in vitro diagnostics...

  15. A Validated High-Throughput Fluorometric Method for Determination of Omeprazole in Quality Control Laboratory via Charge Transfer Sensitized Fluorescence.

    Science.gov (United States)

    Mahmoud, Ashraf M; Ahmed, Sameh A

    2016-03-01

    A high-throughput 96-microwell plate fluorometric method was developed and validated to determine omeprazole (OMZ) in its dosage forms. The method was based on the charge-transfer (CT) sensitized fluorescence reaction of OMZ with 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone (DDQ). This fluorescence reaction provided a new approach for simple, sensitive and selective determinations of OMZ in pharmaceutical preparations. In the present method, the fluorescence reaction was carried out in 96-microwell plates as reaction vessels in order to increase the automation of the methodology and the efficiency of its use in quality control laboratories. All factors affecting the fluorescence reaction were carefully studied and the conditions were optimized. The stoichiometry of the fluorescence reaction between OMZ and DDQ was determined and the reaction mechanism was suggested. Under the optimum conditions, the linear range was 100-6000 ng/ml with the lowest LOD of 33 ng/ml. Analytical performance of the proposed assay, in terms of accuracy and precision, was statistically validated and the results were satisfactory; RSD was <2.6 % and the accuracy was 98.6-101.6 %. The method was successfully applied to the analysis of OMZ in its dosage forms; the recovery values were 98.26-99.60 ± 0.95-2.22 %. The developed methodology may provide a safer, automated and economic tool for the analysis of OMZ in quality control laboratories.

  16. Fluorescence quenching and spectrophotometric methods for the determination of daunorubicin with meso-tera (4-sulphophenyl) porphyrin as probe.

    Science.gov (United States)

    Tian, Jing; Liu, Shaopu; Liu, Zhongfang; Yang, Jidong; Zhu, Jinghui; Qiao, Man; Hu, Xiaoli

    2014-01-01

    In this work, a synthetic meso-tera (4-sulfophenyl) porphyrin (TPPS4) was used as a probe to determine daunorubicin (DNR) by fluorescence quenching and spectrophotometric methods. At pH 4.6 potassium acid phthalate-NaOH buffer solution, a 1:1 complex of DNR interacted with TPPS4 formed via the electrostatic attractions and hydrophobic interactions, thus resulted in TPPS4 fluorescence quenching and absorption spectra change. The maximum excitation wavelength (λex) and the maximum emission wavelength (λem) are 435 nm and 672 nm, respectively. The fluorescence quenching values (ΔF) are the good linear relationship to the concentration of DNR in the range of 0.8-6.0 mgL(-1). The method exhibits high sensitivity with the detection limit (3σ) being 27.0 ng mL(-1). Meanwhile, a decrease of absorbance is detected at 433 nm with the appearance of a new absorption peak at 420 nm. The optimum reaction conditions, influencing factors and the effect of coexisting substances have been investigated in our experiment. The results showed that the method had a good selectivity and could be applied to determine DNR in serum and urine samples. In addition, the combine ratio between DNR and TPPS4 was measured and the charge distribution before and after reaction was calculated by quantum chemistry calculation AM1 method. The type of fluorescence quenching was discussed by the absorption spectra change, Stern-Volmer plots and fluorescence lifetime determination.

  17. A new airborne laser-induced fluorescence instrument for in situ detection of formaldehyde throughout the troposphere and lower stratosphere

    OpenAIRE

    M. Cazorla; Wolfe, G. M.; S. A. Bailey; Swanson, A. K.; H. L. Arkinson; Hanisco, T. F.

    2015-01-01

    The NASA In Situ Airborne Formaldehyde (ISAF) instrument is a high-performance laser-based detector for gas-phase formaldehyde (HCHO). ISAF uses rotational-state specific laser excitation at 353 nm for laser-induced fluorescence (LIF) detection of HCHO. A number of features make ISAF ideal for airborne deployment, including (1) a compact, low-maintenance fiber laser, (2) a single-pass design for stable signal response, (3) a straightforward inlet design, and (4) a stand-alon...

  18. Classification and calibration of organic matter fluorescence data with multiway analysis methods and artificial neural networks: an operational tool for improved drinking water treatment

    OpenAIRE

    Bieroza, Magdalena; Baker, Andy; Bridgeman, John

    2010-01-01

    Fluorescence spectroscopy enables fast and sensitive analysis of environmental samples containing various organic matter constituents. However, to retrieve valuable information from fluorescence spectra, robust techniques for data analysis should be employed. Here, different multivariate analysis methods and artificial neural networks (ANNs) were applied for decomposition and calibration of fluorescence excitation–emission matrices (EEMs). This is the first paper summarizing the application o...

  19. Application of fluorescent microscopy and cascade filtration methods for analysis of soil microbial community

    Science.gov (United States)

    Ivanov, Konstantin; Pinchuk, Irina; Gorodnichev, Roman; Polyanskaya, Lubov

    2016-04-01

    Methods establishment of soil microbial cells size estimation called from the importance of current needs of research in microbial ecology. Some of the methods need to be improved for more detailed view of changes happen in microbiome of terrestrial ecosystems. The combination of traditional microscopy methods, fluorescence and filtration in addition to cutting-edge DNA analysis gives a wide range of the approaches for soil microbial ecologists in their research questions. In the most of the cases the bacterial cells size is limited of the natural conditions such as lack of nutrients or stress factors due to heterogeneity of soil system. In the samples of soils, lakes and rivers sediments, snow and rain water the bacterial cells were detected minimally of 0.2 microns. We established the combination of the cascade filtration and fluorescent microscopy for complex analysis of different terrestrial ecosystems and various soil types. Our modification based on the use of successively filtered soil suspension for collection of microbes by the membrane pores decrease. Combination with fluorescence microscopy and DNA analysis via FISH method gave the presentation of microbial interactions and review of ecological strategies of soil microorganisms. Humus horizons of primitive arctic soil were the most favorable for bacterial growth. Quantified biomass of soil bacteria depends on the dominance of cells with specific dimensions caused of stress factors. The average bacterial size of different soil varied from 0.23 to 0.38 microns, however in humus horizons of arctic soil we detected the contrast dominance of the bigger bacterial cells sized of 1.85 microns. Fungi in this case contributed to increase the availability of organic matter for bacteria because the fungal mycelium forms the appreciable part of microbial biomass of primitive arctic soil. The dominant content of bigger bacterial cells in forest and fallow soil as well as the opposite situation in arable soils caused

  20. Magnetic core-shell fluorescent pH ratiometric nanosensor using a Stoeber coating method

    Energy Technology Data Exchange (ETDEWEB)

    Lapresta-Fernandez, A., E-mail: lapresta@ugr.es [Institute of Physical Chemistry, Friedrich-Schiller-University Jena, Lessingstrasse 10, 07743 Jena (Germany); Instituto de Ciencia de Materiales de Sevilla, centro mixto CSIC-Univ. Sevilla, Avda. Americo Vespucio 49, 41092 Sevilla (Spain); Doussineau, T. [Institute of Physical Chemistry, Friedrich-Schiller-University Jena, Lessingstrasse 10, 07743 Jena (Germany); Universite Lyon 1, CNRS, UMR 5579, LASIM, F-69622 Villeurbanne (France); Moro, A.J. [Institute of Physical Chemistry, Friedrich-Schiller-University Jena, Lessingstrasse 10, 07743 Jena (Germany); REQUIMTE, Departamento de Quimica, Faculdade de Ciencias e Tecnologia, Universidade Nova de Lisboa, 2829-516 Caparica (Portugal); Dutz, S. [Institute of Photonic Technology, Department of Nano Biophotonics, Jena (Germany); Steiniger, F. [Center for Electron Microscopy of the Medical Faculty, Jena (Germany); Mohr, G.J. [Fraunhofer Research Institution for Modular Solid State Technologies, Department of Polytronic Systems, Workgroup Sensor Materials, Josef-Engert-Strasse 13, D-93053 Regensburg (Germany)

    2011-11-30

    Highlights: Black-Right-Pointing-Pointer Architecture combination of magnetic core with two fluorescence silica shells. Black-Right-Pointing-Pointer Both shells properly functionalized which develops ratiometric pH measurements. Black-Right-Pointing-Pointer Reference dye does not change significantly ({approx}1.9%) by modifying the pH. Black-Right-Pointing-Pointer Sensitivity range between 2.0% and 4.9% and a few seconds of response time. Black-Right-Pointing-Pointer One month stability with a signal variation of 4.3%. - Abstract: We describe the use of a modified Stoeber method for coating maghemite ({gamma}-Fe{sub 2}O{sub 3}) nanocrystals with silica shells in order to built magnetic fluorescent sensor nanoparticles in the 50-70 nm diameter range. In detail, the magnetic cores were coated by two successive silica shells embedding two fluorophores (two different silylated dye derivatives), which allows for ratiometric pH-measurements in the pH range 5-8. Silica coated magnetic nanoparticles were prepared using maghemite nanocrystals as cores (5-10 nm in diameter) coated by tetraethoxyorthosilicate via hydrolysis/condensation in ethanol, catalyzed by ammonia. In the inner shell was covalently attached a sulforhodamine B, which was used as a reference dye; while a pH-sensitive fluorescein was incorporated into the outer shell. Once synthesized, the particles were characterized in terms of morphology, size, composition and magnetization, using dynamic light scattering (DLS), transmission electron microscopy (TEM), X-ray diffraction (XRD) and vibrating sample magnetometry (VSM). TEM analysis showed the nanoparticles to be very uniform in size. Wide-angle X-ray diffractograms showed, for uncoated as well as coated nanoparticles, typical peaks for the spinel structure of maghemite at the same diffraction angle, with no structural changes after coating. When using VSM, we obtained the magnetization curves of the resulting nanoparticles and the typical magnetization

  1. Aggregation-induced fluorescence behavior of triphenylamine-based Schiff bases: the combined effect of multiple forces.

    Science.gov (United States)

    Yang, Mingdi; Xu, Dongling; Xi, Wengang; Wang, Lianke; Zheng, Jun; Huang, Jing; Zhang, Jingyan; Zhou, Hongping; Wu, Jieying; Tian, Yupeng

    2013-10-18

    Eight triphenylamine (TPA)-based Schiff bases that exhibit different aggregation-induced emission (AIE) or aggregation-caused quenching (ACQ) behavior in tetrahydrofuran (THF)/water mixtures have been synthesized and characterized. The photophysical properties in solution, aqueous suspension, film, and the crystalline state along with their relationships were comparatively investigated. The single-crystal structures of 1-8 indicate that compact π···π stacking or excimers induce fluorescence quenching of 1, 2, 5, and 7. However, the existence of J aggregates or multiple intra- and intermolecular interactions restrict the intramolecular vibration and rotation, enabling compounds 3, 4, 6, and 8 to exhibit good AIE character. The size and growth process of particles with different water fractions were studied using scanning electron microscopy, which demonstrated that smaller uniformly dispersed nanoparticles in the THF/water mixtures favor fluorescence emission. The above results suggest that the combined effects of multiple forces caused by structural variation have a great influence on their molecular packing, electronic structure, and aggregation-induced fluorescence properties. In addition, piezofluorochromic experiments verified the potential applications of 4 and 6.

  2. Spectral resolved Measurement of the Nitrogen Fluorescence Emissions in Air induced by Electrons

    CERN Document Server

    Waldenmaier, Tilo; Klages, Hans

    2007-01-01

    For the calorimetric determination of the primary energy of extensive air showers, measured by fluorescence telescopes, a precise knowledge of the conversion factor (fluorescence yield) between the deposited energy in the atmosphere and the number of emitted fluorescence photons is essential. The fluorescence yield depends on the pressure and the temperature of the air as well as on the water vapor concentration. Within the scope of this work the fluorescence yield for the eight strongest nitrogen emission bands between 300 nm and 400 nm has been measured using electrons from a Sr-90 source with energies between 250 keV and 2000 keV. Measurements have been performed in dry air, pure nitrogen, and a nitrogen-oxygen mixture at pressures ranging from 2 hPa to 990 hPa. Furthermore the influence of water vapor has been studied. A new approach for the parametrization of the fluorescence yield was used to analyze the data, leading to a consistent description of the fluorescence yield with a minimal set of parameters...

  3. Measurement of the pressure dependence of air fluorescence emission induced by electrons

    CERN Document Server

    Ave, M; Buonomo, B; Busca, N; Cazon, L; Chemerisov, S D; Conde, M E; Crowell, R A; Di Carlo, P; Di Giulio, C; Doubrava, M; Esposito, A; Facal, P; Franchini, F J; Horandel, J; Hrabovsky, M; Iarlori, M; Kasprzyk, T E; Keilhauer, B; Klages, H; Kleifges, M; Kuhlmann, S; Mazzitelli, G; Nozka, L; Obermeier, A; Palatka, M; Petrera, S; Privitera, P; Rídky, J; Rizi, V; Rodríguez, G; Salamida, F; Schovanek, P; Spinka, H; Strazzeri, E; Ulrich, A; Yusof, Z M; Vacek, V; Valente, P; Verzi, V; Waldenmaier, T

    2007-01-01

    The fluorescence detection of ultra high energy (> 10^18 eV) cosmic rays requires a detailed knowledge of the fluorescence light emission from nitrogen molecules, which are excited by the cosmic ray shower particles along their path in the atmosphere. We have made a precise measurement of the fluorescence light spectrum excited by MeV electrons in dry air. We measured the relative intensities of 34 fluorescence bands in the wavelength range from 284 to 429 nm with a high resolution spectrograph. The pressure dependence of the fluorescence spectrum was also measured from a few hPa up to atmospheric pressure. Relative intensities and collisional quenching reference pressures for bands due to transitions from a common upper level were found in agreement with theoretical expectations. The presence of argon in air was found to have a negligible effect on the fluorescence yield. We estimated that the systematic uncertainty on the cosmic ray shower energy due to the pressure dependence of the fluorescence spectrum i...

  4. Containerless Atomic-Fluorescence Property Measurements

    Science.gov (United States)

    Nordine, P.; Schiffman, R.; Walker, C.

    1987-01-01

    Report describes studies conducted to establish and verify use of laser-induced fluorescence in monitoring and controlling high-temperature containerless processes. Specimens levitated by gas jets or electromagnetic fields and heated by laser beams or electromagnetic induction while being irradiated and detected by fluorescence technique. Makes quantitative and qualitative comparisons among three new methods of temperature measurement; all rely on laser-induced fluorescence. One method gas-density thermometry with seed gas. Other two methods involve measurements of velocities of evaporating atoms or of population ratios of different electronic states.

  5. Measurement of OH reactivity by laser flash photolysis coupled with laser-induced fluorescence spectroscopy

    Science.gov (United States)

    Stone, Daniel; Whalley, Lisa K.; Ingham, Trevor; Edwards, Peter M.; Cryer, Danny R.; Brumby, Charlotte A.; Seakins, Paul W.; Heard, Dwayne E.

    2016-07-01

    OH reactivity (k'OH) is the total pseudo-first-order loss rate coefficient describing the removal of OH radicals to all sinks in the atmosphere, and is the inverse of the chemical lifetime of OH. Measurements of ambient OH reactivity can be used to discover the extent to which measured OH sinks contribute to the total OH loss rate. Thus, OH reactivity measurements enable determination of the comprehensiveness of measurements used in models to predict air quality and ozone production, and, in conjunction with measurements of OH radical concentrations, to assess our understanding of OH production rates. In this work, we describe the design and characterisation of an instrument to measure OH reactivity using laser flash photolysis coupled to laser-induced fluorescence (LFP-LIF) spectroscopy. The LFP-LIF technique produces OH radicals in isolation, and thus minimises potential interferences in OH reactivity measurements owing to the reaction of HO2 with NO which can occur if HO2 is co-produced with OH in the instrument. Capabilities of the instrument for ambient OH reactivity measurements are illustrated by data collected during field campaigns in London, UK, and York, UK. The instrumental limit of detection for k'OH was determined to be 1.0 s-1 for the campaign in London and 0.4 s-1 for the campaign in York. The precision, determined by laboratory experiment, is typically < 1 s-1 for most ambient measurements of OH reactivity. Total uncertainty in ambient measurements of OH reactivity is ˜ 6 %. We also present the coupling and characterisation of the LFP-LIF instrument to an atmospheric chamber for measurements of OH reactivity during simulated experiments, and provide suggestions for future improvements to OH reactivity LFP-LIF instruments.

  6. Consistency Between Sun-Induced Chlorophyll Fluorescence and Gross Primary Production of Vegetation in North America

    Science.gov (United States)

    Zhang, Yao; Xiao, Xiangming; Jin, Cui; Dong, Jinwei; Zhou, Sha; Wagle, Pradeep; Joiner, Joanna; Guanter, Luis; Zhang, Yongguang; Zhang , Geli; Qin, Yuanwei; Wang, Jie; Moore, Berrien, III

    2016-01-01

    Accurate estimation of the gross primary production (GPP) of terrestrial ecosystems is vital for a better understanding of the spatial-temporal patterns of the global carbon cycle. In this study,we estimate GPP in North America (NA) using the satellite-based Vegetation Photosynthesis Model (VPM), MODIS (Moderate Resolution Imaging Spectrometer) images at 8-day temporal and 500 meter spatial resolutions, and NCEP-NARR (National Center for Environmental Prediction-North America Regional Reanalysis) climate data. The simulated GPP (GPP (sub VPM)) agrees well with the flux tower derived GPP (GPPEC) at 39 AmeriFlux sites (155 site-years). The GPP (sub VPM) in 2010 is spatially aggregated to 0.5 by 0.5-degree grid cells and then compared with sun-induced chlorophyll fluorescence (SIF) data from Global Ozone Monitoring Instrument 2 (GOME-2), which is directly related to vegetation photosynthesis. Spatial distribution and seasonal dynamics of GPP (sub VPM) and GOME-2 SIF show good consistency. At the biome scale, GPP (sub VPM) and SIF shows strong linear relationships (R (sup 2) is greater than 0.95) and small variations in regression slopes ((4.60-5.55 grams Carbon per square meter per day) divided by (milliwatts per square meter per nanometer per square radian)). The total annual GPP (sub VPM) in NA in 2010 is approximately 13.53 petagrams Carbon per year, which accounts for approximately 11.0 percent of the global terrestrial GPP and is within the range of annual GPP estimates from six other process-based and data-driven models (11.35-22.23 petagrams Carbon per year). Among the seven models, some models did not capture the spatial pattern of GOME-2 SIF data at annual scale, especially in Midwest cropland region. The results from this study demonstrate the reliable performance of VPM at the continental scale, and the potential of SIF data being used as a benchmark to compare with GPP models.

  7. Pressure broadening of atomic oxygen two-photon absorption laser induced fluorescence

    Science.gov (United States)

    Marinov, Daniil; Drag, Cyril; Blondel, Christophe; Guaitella, Olivier; Golda, Judith; Klarenaar, Bart; Engeln, Richard; Schulz-von der Gathen, Volker; Booth, Jean-Paul

    2016-12-01

    Atomic oxygen, considered to be a determining reactant in plasma applications at ambient pressure, is routinely detected by two-photon absorption laser induced fluorescence (TALIF). Here, pressure broadening of the (2p 4 3 P 2  →  3p 3 P J=0,1,2) two-photon transition in oxygen atoms was investigated using a high-resolution TALIF technique in normal and Doppler-free configurations. The pressure broadening coefficients determined were {γ{{\\text{O}2}}}   =  0.40  ±  0.08  cm-1/bar for oxygen molecules and {γ\\text{He}}   =  0.46  ±  0.03 cm-1/bar for helium atoms. These correspond to pressure broadening rate constants k\\text{PB}{{\\text{O}2}}   =  9 · 10-9 cm3 s-1 and k\\text{PB}\\text{He}   =  4 · 10-9 cm3 s-1, respectively. The well-known quenching rate constants of O(3p 3 P J ) by O2 and He are at least one order of magnitude smaller, which signifies that non-quenching collisions constitute the main line-broadening mechanism. In addition to providing new insights into collisional processes of oxygen atoms in electronically excited 3p 3 P J state, reported pressure broadening parameters are important for quantification of oxygen TALIF line profiles when both collisional and Doppler broadening mechanisms are important. Thus, the Doppler component (and hence the temperature of oxygen atoms) can be accurately determined from high resolution TALIF measurements in a broad range of conditions.

  8. Standoff laser-induced fluorescence of suspensions from different bacterial strains

    Science.gov (United States)

    Duschek, Frank; Walter, Arne; Fellner, Lea; Grünewald, Karin; Pargmann, Carsten; Handke, Jürgen; Tomaso, Herbert

    2016-10-01

    Biological hazardous substances like certain fungi and bacteria represent a high risk for the broad public if fallen into wrong hands. Incidents based on bio agents are commonly considered to have incalculable and complex consequences for first responders and people. The impact of such an event can be minimized by a combination of different sensor technologies that have been developed to detect bio-threats and to gather information after an incident. Sensors for bio-agents can be grouped into two categories. Sampling devices collect material from locations supposed to be contaminated, and they are able to identify biological material with high sensitivity and selectivity. However, these point sensors need to be positioned correctly in advance of an attack, and moving sources of biological material cannot be tracked. A different approach is based on optical standoff detection. For biological samples laser induced florescence (LIF) has been proven to get real time data on location and type of hazards without being in contact with the suspicious substance. This work is based on a bio-detector developed at the DLR Lampoldshausen. The LIF detection has been designed for outdoor operation at standoff distances from 20 m up to more than 100 m. The detector acquires LIF spectral data for two different excitation wavelengths (280 and 355 nm) as well as time resolved information for the fluorescence decay which can be used to classify suspicious samples. While the classification device had been trained on uncritical samples (like amino acids, NADH, yeast, chemicals, oils), this work presents the progress to more relevant, living bacteria of different strains. The low risk and non-pathogenic bacteria Bacillus thuringensis, Bacillus atrophaeus, Bacillus subtilis, Brevibacillus brevis, Micrococcus luteus, Oligella urethralis, Paenibacillus polymyxa and Escherichia coli (K12) have been investigated with the above set-up at both excitation wavelengths

  9. Ex vivo optical coherence tomography and laser induced fluorescence spectroscopy imaging of murine gastrointestinal tract

    Science.gov (United States)

    Hariri, Lida; Tumlinson, Alexandre R.; Wade, Norman; Besselsen, David; Utzinger, Urs; Gerner, Eugene; Barton, Jennifer

    2005-04-01

    Optical Coherence Tomography (OCT) and Laser Induced Fluorescence Spectroscopy (LIF) have separately been found to have clinical potential in identifying human gastrointestinal (GI) pathologies, yet their diagnostic capability in mouse models of human disease is unknown. We combine the two modalities to survey the GI tract of a variety of mouse strains and sample dysplasias and inflammatory bowel disease (IBD) of the small and large intestine. Segments of duodenum and lower colon 2.5 cm in length and the entire esophagus from 10 mice each of two colon cancer models (ApcMin and AOM treated A/J) and two IBD models (Il-2 and Il-10) and 5 mice each of their respective controls were excised. OCT images and LIF spectra were obtained simultaneously from each tissue sample within 1 hour of extraction. Histology was used to classify tissue regions as normal, Peyer"s patch, dysplasia, adenoma, or IBD. Features in corresponding regions of OCT images were analyzed. Spectra from each of these categories were averaged and compared via the student's t-test. Features in OCT images correlated to histology in both normal and diseased tissue samples. In the diseased samples, OCT was able to identify early stages of mild colitis and dysplasia. In the sample of IBD, the LIF spectra displayed unique peaks at 635nm and 670nm, which were attributed to increased porphyrin production in the proliferating bacteria of the disease. These peaks have the potential to act as a diagnostic for IBD. OCT and LIF appear to be useful and complementary modalities for imaging mouse models.

  10. Evaluation of a direct fluorescent antibody staining method for rapid identification of members of the bacteroides fragilis group.

    Science.gov (United States)

    DeGirolami, P C; Mepani, C P

    1981-07-01

    A direct fluorescent antibody test kit (Fluorotec-F, Pfizer Inc., New York, New York) designed for rapid identification of members of the Bacteroides fragilis group (BFG) was evaluated. Tested were 228 clinical specimens (144 direct smears of clinical material, 14 smears of positive blood cultures, and 70 smears of colonies isolated from clinical material) and 49 reference strains of anaerobic bacteria, including 23 members of the BFG. Fluorotec-F detected 68 of 69 (98.5%) members of the BFG, including 55 B. fragilis, 12 B. thetaiotaomicron, and two B. ovatus, identified by cultural methods in all clinical specimens. Three specimens that yielded B. uniformis also fluoresced. Three specimens fluoresced but failed to yield members of the BFG or B. uniformis on culture. Of the 49 reference strains tested, all strains of B. fragilis, B. thetaiotaomicron, nd B. uniformis tested were detected by Fluorotec-F, but only five of a total of 14B. vulgatus, B. distasonis, and B. ovatus tested fluoresced. Of the 25 reference strains of anaerobic bacteria not belonging to the BFG, none fluoresced except for two strains of B. eggerthii. Direct fluorescent antibody staining of smears of clinical specimens suitable for anaerobic culture is a valuable tool for rapid detection of B. fragilis infections.

  11. Novel Method for Indirect Determination of Iodine in Marine Products by Atomic Fluorescence Spectrometry

    Institute of Scientific and Technical Information of China (English)

    LU Jian-ping; TAN Fang-wei; TANG Qiong; JIANG Tian-cheng

    2013-01-01

    A method for the determination of iodine based upon compound H2HgI4,formed between I-and Hg2+ in nitric acid and extracted in methyl isobutyl ketone(MIBK),was developed via atomic fluorescence spectrometry(AFS).After the compound is reduced with potassium borohydrid(KBH4),the resultant mercury vapor was injected into the instrument and iodine was,therefore,indirectly determined.Experimental parameters such as the conditions of extraction reagents,aqueous phase acidity,elemental mercury diffusion temperature in a vial and other factors were investigated and optimized.Under the optimum experimental conditions,this method shows a detection limit of 0.038 μg/L iodine and a linear relationship between 0.04-20 μg/L.The