Donor-acceptor-pair emission in fluorescent 4H-SiC grown by PVT method

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

Liu, Xi, E-mail: liuxi@mail.sic.ac.cn; Zhuo, Shi-Yi; Gao, Pan

Fluorescent SiC, which contains donor and acceptor impurities with optimum concentrations, can work as a phosphor for visible light emission by donor-acceptor-pair (DAP) recombination. In this work, 3 inch N-B-Al co-doped fluorescent 4H-SiC crystals are prepared by PVT method. The p-type fluorescent 4H-SiC with low aluminum doping concentration can show intensive yellow-green fluorescence at room temperature. N-B DAP peak wavelength shifts from 578nm to 525nm and weak N-Al DAP emission occurred 403/420 nm quenches, when the temperature increases from 4K to 298K. The aluminum doping induces higher defect concentration in the fluorescent crystal and decreases optical transmissivity of the crystalmore » in the visible light range. It triggers more non-radiative recombination and light absorption losses in the crystal.« less

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

NASA Astrophysics Data System (ADS)

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.

Detection and evaluation of normal and malignant cells using laser-induced fluorescence spectroscopy.

PubMed

Khosroshahi, Mohamad E; Rahmani, Mahya

2012-01-01

The aim of this research is to study the normalized fluorescence spectra (intensity variations and area under the fluorescence signal), relative quantum yield, extinction coefficient and intracellular properties of normal and malignant human bone cells. Using Laser-Induced Fluorescence Spectroscopy (LIFS) upon excitation of 405 nm, the comparison of emission spectra of bone cells revealed that fluorescence intensity and the area under the spectra of malignant bone cells was less than that of normal. In addition, the area ratio and shape factor were changed. We obtained two emission bands in spectra of normal cells centered at about 486 and 575 nm and for malignant cells about 482 and 586 nm respectively, which are most likely attributed to NADH and riboflavins. Using fluorescein sodium emission spectrum, the relative quantum yield of bone cells is numerically determined.

Construction of a ratiometric fluorescent probe with an extremely large emission shift for imaging hypochlorite in living cells

NASA Astrophysics Data System (ADS)

Song, Xuezhen; Dong, Baoli; Kong, Xiuqi; Wang, Chao; Zhang, Nan; Lin, Weiying

2018-01-01

Hypochlorite is one of the important reactive oxygen species (ROS) and plays critical roles in many biologically vital processes. Herein, we present a unique ratiometric fluorescent probe (CBP) with an extremely large emission shift for detecting hypochlorite in living cells. Utilizing positively charged α,β-unsaturated carbonyl group as the reaction site, the probe CBP itself exhibited near-infrared (NIR) fluorescence at 662 nm, and can display strong blue fluorescence at 456 nm when responded to hypochlorite. Notably, the extremely large emission shift of 206 nm could enable the precise measurement of the fluorescence peak intensities and ratios. CBP showed high sensitivity, excellent selectivity, desirable performance at physiological pH, and low cytotoxicity. The bioimaging experiments demonstrate the biological application of CBP for the ratiometric imaging of hypochlorite in living cells.

Foliar Reflectance and Fluorescence Responses for Plants Under Nitrogen Stress Determined with Active and Passive Systems

NASA Technical Reports Server (NTRS)

Middleton, E. M.; McMurtrey, J. E.; Campbell, P. K. Entcheva; Corp, L. A.; Butcher, L. M.; Chappelle, E. W.

2003-01-01

Vegetation productivity is driven by nitrogen (N) availability in soils. Both excessive and low soil N induce physiological changes in plant foliage. In 2001, we examined the use of spectral fluorescence and reflectance measurements to discriminate among plants provided different N fertilizer application rates: 20%, 50%, 100% and 150% of optimal N levels. A suite of optical, fluorescence, and biophysical measurements were collected on leaves from field grown corn (Zea mays L.) and soybean plants (Glycine max L.) grown in pots (greenhouse + ambient sunlight daily). Three types of steady state laser-induced fluorescence measurements were made on adaxial and abaxial surfaces: 1) fluorescence images in four 10 nm bands (blue, green, red, far-red) resulting from broad irradiance excitation; 2) emission spectra (5 nm resolution) produced by excitation at single wavelengths (280,380 or 360, and 532 nm); and 3) excitation spectra (2 nm resolution), with emission wavelengths fixed at wavelengths centered on selected solar Fraunhofer lines (532,607,677 and 745 nm). Two complementary sets of high resolution (less than 2 nm) optical spectra were acquired for both adaxial and abaxial leaf surfaces: 1) optical properties (350-2500 nm) for reflectance, transmittance, and absorptance; and 2) reflectance spectra (500-1000 nm) acquired with and without a short pass filter at 665 nm to determine the fluorescence contribution to apparent reflectance in the 650-750 spectrum, especially at the 685 and 740 nm chlorophyll fluorescence (ChIF) peaks. The strongest relationships between foliar chemistry and optical properties were demonstrated for C/N content and two optical parameters associated with the red edge inflection point. Select optical properties and ChIF parameters were highly correlated for both species. A significant contribution of ChIF to apparent reflectance was observed, averaging 10-25% at 685 nm and 2 - 6% at 740 nm over all N treatments. Discrimination of N treatment groups was possible with specific fluorescence band ratios (e.g., F740/F525 obtained with 380EX). From all measurements assessing fluorescence, higher ChIF and blue/green emissions were measured from the abaxial leaf surfaces; Abaxial surfaces also produced higher reflectances in the 400-800 nm spectrum. Fluorescence information collected in Fraunhofer regions located on the shoulders of ChIF features compared favorably with peak emissions. This supports the potential capability of a future space-born interferometer sensor to capture plant canopy fluorescence.

Development of in-vitro models to elucidate mechanisms of intrinsic cellular and tissue fluorescence

NASA Astrophysics Data System (ADS)

Savage, Howard E.; Kolli, Venkateswara; Saha, Sanjoy; Zhang, Jian C.; Glasgold, Mark; Sacks, Peter G.; Alfano, Robert R.; Schantz, Stimson P.

1995-04-01

In vitro cell model systems have been used to study the mechanisms of intrinsic cellular and tissue fluorescence as a potential biomarker for cancer. Phenotypic characteristics of cancer that are different from normal tissue include changes in histoarchitecture, proliferation rates and differentiation. a nitrosmethlybenzylamine (NMBA)/rat esophageal carcinogenesis model (NMBA), a transforming growth factor beta (TGF- (beta) )/normal epithelial cell model, and a retinoic acid (RA)/multicellular tumor spheroid model (RAMTS) were used to assess fluorescence changes associated respectively with changes in histoarchitecture, proliferation rates and differentiation. A xenon based fluorescence spectrophotometer (Mediscience Corp.) was used to collect excitation and emission spectra. Two excitation scans ((lambda) Ex 200-360 nm, (lambda) Em 380 nm; (lambda) Ex 240-430 nm, (lambda) Em 450 nm) and two emission scans ((lambda) Ex 300 nm, (lambda) Em 320-580 nm; (lambda) Ex 340 nm, (lambda) Em 360-660 nm) were used to analyze the three model systems. Using the NMBA model. Differences were seen in the excitation scan ((lambda) Ex 200-360 nm, (lambda) Em 380 nm) and the emission scan ((lambda) Ex 340 nm, (lambda) Em 360-660 nm) when normal rat esophageal tissue was compared to hyperplastic and tumor tissue. In the (TGF-(beta) ) model, differences were seen in the excitation scan ((lambda) Ex 240-430 nm, (lambda) Em 450 nm) when comparing proliferation slowed (TGF-(beta) treated) epithelial cells to their untreated controls. In the RAMTS model, differences were seen with all four scans when RA treated multicellular tumor spheroids (nondifferentiating) were compared to untreated control cells (differentiating). The data indicate that fluorescence changes seen in these model systems may relate to changes in histoarchitecture, proliferation rates and differentiation. Their relationship to in vivo fluorescence changes seen in cancer patients remains to be elucidated.

Real-time monitoring of river water quality using in-line continuous acquisition of fluorescence excitation and emission matrices

NASA Astrophysics Data System (ADS)

Carstea, E.; Baker, A.; Johnson, R.; Reynolds, D. M.

2009-12-01

In-line fluorescence EEM monitoring has been performed over an eleven-day period for Bournbrook River, Birmingham, UK. River water was diverted to a portable laboratory via a continuous flow pump and filter system. Fluorescence excitation-emission matrices data was recorded every 3 minutes using a flow cell (1cm pathlength) coupled to a fiber optic probe. This real-time fluorescence EEM data (Excitation, 225-400 nm at 5 nm steps, emission, 280-500 nm at 2 nm steps) was collected 'in-line'and directly compared with the spectrophotometric properties and physical and chemical parameters of river water samples collected off-line at known time intervals. Over the monitoring period, minor pollution pulses from cross connections were detected and identified hourly along with a random diesel pollution event. This work addresses the practicalities of measuring and detecting fluorescence EEM in the field and discusses the potential of this technological approach for further understanding important hydrological and biogeochemical processes. Problems associated with fouling and system failure are also reported. Example of the data generated from the continuous fluorescence EEM monitoring.

Optical spectroscopy of the bladder washout fluid to optimize fluorescence cystoscopy with Hexvix®.

PubMed

Martoccia, Carla; Zellweger, Matthieu; Lovisa, Blaise; Jichlinski, Patrice; van den Bergh, Hubert; Wagnières, Georges

2014-09-01

Fluorescence cystoscopy enhances detection of early bladder cancer. Water used to inflate thebladder during the procedure rapidly contains urine, which may contain fluorochromes. This frequently degradesfluorescence images. Samples of bladder washout fluid (BWF) or urine were collected (15 subjects). We studiedtheir fluorescence properties and assessed changes induced by pH (4 to 9) and temperature (15°C to 41°C).A typical fluorescence spectrum of BWF features a main peak (excitation/emission: 320∕420 nm, FWHM =50∕100 nm) and a weaker (5% to 20% of main peak intensity), secondary peak (excitation/emission: 455∕525 nm, FWHM = 80∕50 nm). Interpatient fluctuations of fluorescence intensity are observed. Fluorescence intensity decreases when temperature increases (max 30%) or pH values vary (max 25%). Neither approach is compatible with clinical settings. Fluorescence lifetime measurements suggest that 4-pyridoxic acid/riboflavin is the most likely molecule responsible for urine’s main/secondary fluorescence peak. Our measurements give an insight into the spectroscopy of the detrimental background fluorescence. This should be included in the optical design of fluorescence cystoscopes. We estimate that restricting the excitation range from 370–430 nm to 395–415 nm would reduce the BWF background by a factor 2.

UV induced visual cues in grasses

PubMed Central

Baby, Sabulal; Johnson, Anil John; Govindan, Balaji; Lukose, Sujith; Gopakumar, Bhaskaran; Koshy, Konnath Chacko

2013-01-01

Grasses are traditionally considered as wind pollinated, however, field observations confirmed frequent insect visits to grass flowers, suggesting insect pollination. Fruit and seed predators inflict heavy losses to cereals and millets during their growth, maturation and storage. The actual factors guiding insects and predators to grass flowers, fruits and seeds are not clear. Here, we report attractive blue fluorescence emissions on grass floral parts such as glumes, lemma, palea, lodicules, staminal filaments, pollens and fruits in ultraviolet (UV) 366 nm, whereas the stigmatic portions were not blue, but red fluorescent. We characterized the blue fluorescent constituent in grass reproductive structures as ferulic acid (FA). Fluorescence spectra of blue-emitting grass floral, seed extracts and isolated FA on excitation at 366 nm showed their emissions at 420–460 nm. We propose these FA-based blue fluorescence emissions in grass reproductive structures as visual cues that attract pollinators, predators and even pests towards them. PMID:24061408

Parametric models to compute tryptophan fluorescence wavelengths from classical protein simulations.

PubMed

Lopez, Alvaro J; Martínez, Leandro

2018-02-26

Fluorescence spectroscopy is an important method to study protein conformational dynamics and solvation structures. Tryptophan (Trp) residues are the most important and practical intrinsic probes for protein fluorescence due to the variability of their fluorescence wavelengths: Trp residues emit in wavelengths ranging from 308 to 360 nm depending on the local molecular environment. Fluorescence involves electronic transitions, thus its computational modeling is a challenging task. We show that it is possible to predict the wavelength of emission of a Trp residue from classical molecular dynamics simulations by computing the solvent-accessible surface area or the electrostatic interaction between the indole group and the rest of the system. Linear parametric models are obtained to predict the maximum emission wavelengths with standard errors of the order 5 nm. In a set of 19 proteins with emission wavelengths ranging from 308 to 352 nm, the best model predicts the maximum wavelength of emission with a standard error of 4.89 nm and a quadratic Pearson correlation coefficient of 0.81. These models can be used for the interpretation of fluorescence spectra of proteins with multiple Trp residues, or for which local Trp environmental variability exists and can be probed by classical molecular dynamics simulations. © 2018 Wiley Periodicals, Inc. © 2018 Wiley Periodicals, Inc.

System and method for controlling depth of imaging in tissues using fluorescence microscopy under ultraviolet excitation following staining with fluorescing agents

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

Levenson, Richard; Demos, Stavros

A method is disclosed for analyzing a thin tissue sample and adapted to be supported on a slide. The tissue sample may be placed on a slide and exposed to one or more different exogenous fluorophores excitable in a range of about 300 nm-200 nm, and having a useful emission band from about 350 nm-900 nm, and including one or more fluorescent dyes or fluorescently labeled molecular probes that accumulate in tissue or cellular components. The fluorophores may be excited with a first wavelength of UV light between about 200 nm-290 nm. An optical system collects emissions from the fluorophoresmore » at a second wavelength, different from the first wavelength, which are generated in response to the first wavelength of UV light, to produce an image for analysis.« less

Spectral dependence of irreversible light-induced fluorescence quenching: Chlorophyll forms with maximal emission at 700-702 and 705-710nm as spectroscopic markers of conformational changes in the core complex.

PubMed

Nematov, Sherzod; Casazza, Anna Paola; Remelli, William; Khuvondikov, Vakhobjon; Santabarbara, Stefano

2017-07-01

The spectral dependence of the irreversible non-photochemical fluorescence quenching associated with photoinhibition in vitro has been comparatively investigated in thylakoid membranes, PSII enriched particles and PSII core complexes isolated from spinach. The analysis of the fluorescence emission spectra of dark-adapted and quenched samples as a function of the detection temperature in the 280-80K interval, indicates that Chlorophyll spectral forms having maximal emission in the 700-702nm and 705-710nm ranges gain relative intensity in concomitance with the establishment of irreversible light-induced quenching, acting thereby as spectroscopic markers. The relative enhancement of the 700-702nm and 705-710nm forms emission could be due either to an increase of their stoichiometric abundance or to their intrinsically low fluorescence quantum yields. These two factors, that can also coexist, need to be promoted by light-induced alterations in chromophore-protein as well as chromophore-chromophore interactions. The bands centred at about 701 and 706nm are also observed in the PSII core complex, suggesting their, at least partial, localisation in proximity to the reaction centre, and the occurrence of light-induced conformational changes in the core subunits. Copyright © 2017 Elsevier B.V. All rights reserved.

Native fluorescence characterization of human liver abnormalities

NASA Astrophysics Data System (ADS)

Ganesan, Singaravelu; Madhuri, S.; Aruna, Prakasa R.; Suchitra, S.; Srinivasan, T. G.

1999-05-01

Fluorescence spectroscopy of intrinsic biomolecules has been extensively used in biology and medicine for the past several decades. In the present study, we report the native fluorescence characteristics of blood plasma from normal human subjects and patients with different liver abnormalities such as hepatitis, leptospirosis, jaundice, cirrhosis and liver cell failure. Native fluorescence spectra of blood plasma -- acetone extract were measured at 405 nm excitation. The average spectrum of normal blood plasma has a prominent emission peak around 464 nm whereas in the case of liver diseased subjects, the primary peak is red shifted with respect to normal. In addition, liver diseased cases show distinct secondary emission peak around 615 nm, which may be attributed to the presence of endogenous porphyrins. The red shift of the prominent emission peak with respect to normal is found to be maximum for hepatitis and minimum for cirrhosis whereas the secondary emission peak around 615 nm was found to be more prominent in the case of cirrhosis than the rest. The ratio parameter I465/I615 is found to be statistically significant (p less than 0.001) in discriminating liver abnormalities from normal.

Raman scattering and red fluorescence in the photochemical transformation of dry tryptophan particles

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

Lai, Chih Wei; Schwab, Mark; Hill, Steven C.

Tryptophan is a fluorescent amino acid common in proteins. Its absorption is largest for wavelengths λ ≲ 290 nm and its fluorescence emissions peak around 300–350 nm, depending upon the local environment. Here we report the observation of red fluorescence near 600 nm emerging from 488-nm continuous-wave (CW) laser photoexcitation of dry tryptophan (Trp) particles. With an excitation intensity below 0.5 kW/cm 2, dry Trp particles yield distinctive Raman scattering peaks in the presence of relatively weak and spectrally broad emissions with λ ~500–700 nm, allowing estimation of particle temperature at low excitation intensities. When the photoexcitation intensity is increasedmore » to 1 kW/cm 2 or more for a few minutes, fluorescence intensity dramatically increases by more than two orders of magnitude. The fluorescence continues to increase in intensity and gradually shift to the red when photoexcitation intensity and the duration of exposure are increased. The resulting products absorb at visible wavelengths and generate red fluorescence with λ ~ 650–800 nm with 633-nm CW laser excitation. In conclusion, we attribute the emergence of orange and red fluorescence in the Trp products to a photochemical transformation that is instigated by weak optical transitions to triplet states in Trp with 488-nm excitation and which may be expedited by a photothermal effect.« less

Raman scattering and red fluorescence in the photochemical transformation of dry tryptophan particles

DOE PAGES

Lai, Chih Wei; Schwab, Mark; Hill, Steven C.; ...

2016-05-19

Tryptophan is a fluorescent amino acid common in proteins. Its absorption is largest for wavelengths λ ≲ 290 nm and its fluorescence emissions peak around 300–350 nm, depending upon the local environment. Here we report the observation of red fluorescence near 600 nm emerging from 488-nm continuous-wave (CW) laser photoexcitation of dry tryptophan (Trp) particles. With an excitation intensity below 0.5 kW/cm 2, dry Trp particles yield distinctive Raman scattering peaks in the presence of relatively weak and spectrally broad emissions with λ ~500–700 nm, allowing estimation of particle temperature at low excitation intensities. When the photoexcitation intensity is increasedmore » to 1 kW/cm 2 or more for a few minutes, fluorescence intensity dramatically increases by more than two orders of magnitude. The fluorescence continues to increase in intensity and gradually shift to the red when photoexcitation intensity and the duration of exposure are increased. The resulting products absorb at visible wavelengths and generate red fluorescence with λ ~ 650–800 nm with 633-nm CW laser excitation. In conclusion, we attribute the emergence of orange and red fluorescence in the Trp products to a photochemical transformation that is instigated by weak optical transitions to triplet states in Trp with 488-nm excitation and which may be expedited by a photothermal effect.« less

Raman scattering and red fluorescence in the photochemical transformation of dry tryptophan particles.

PubMed

Lai, Chih Wei; Schwab, Mark; Hill, Steven C; Santarpia, Joshua; Pan, Yong-Le

2016-05-30

Tryptophan is a fluorescent amino acid common in proteins. Its absorption is largest for wavelengths λ ≲ 290 nm and its fluorescence emissions peak around 300-350 nm, depending upon the local environment. Here we report the observation of red fluorescence near 600 nm emerging from 488-nm continuous-wave (CW) laser photoexcitation of dry tryptophan (Trp) particles. With an excitation intensity below 0.5 kW/cm², dry Trp particles yield distinctive Raman scattering peaks in the presence of relatively weak and spectrally broad emissions with λ ∼500-700 nm, allowing estimation of particle temperature at low excitation intensities. When the photoexcitation intensity is increased to 1 kW/cm² or more for a few minutes, fluorescence intensity dramatically increases by more than two orders of magnitude. The fluorescence continues to increase in intensity and gradually shift to the red when photoexcitation intensity and the duration of exposure are increased. The resulting products absorb at visible wavelengths and generate red fluorescence with λ ∼ 650-800 nm with 633-nm CW laser excitation. We attribute the emergence of orange and red fluorescence in the Trp products to a photochemical transformation that is instigated by weak optical transitions to triplet states in Trp with 488-nm excitation and which may be expedited by a photothermal effect.

The modification of spectral characteristics of cytostatics by optical beams

NASA Astrophysics Data System (ADS)

Pascu, Mihail Lucian; Brezeanu, Mihail; Carstocea, Benone D.; Voicu, Letitia; Gazdaru, Doina M.; Smarandache, Adriana A.

2004-10-01

Besides the biochemical action of methotrexate (MTX) and 5-fluorouracil (FU) their effect in destroying cancer tumours could be enhanced by exposure to light at different doses. Absorption, excitation and emission spectra of 10-4M - 10-5M MTX solutions in natural saline and sodium hydroxide at pH = 8.4 were measured, while their exposure to coherent and uncoherent light in the visible and near ultraviolet (UV) spectral ranges was made (Hg lamps and Nitrogen pulsed laser radiation were used). Absorption spectra exhibit spectral bands in the range 200 nm - 450 nm. The 200 - 450 nm excitation spectra were measured with emission centered on 470 nm; MTX fluorescence excitation was measured at 390 nm and the emission was detected between 400 nm and 600 nm showing a maximum at 470 nm. Spectra modifications, nonlinearly depending on exposure time (varying from 1 min to 20 min), evidenced MTX photo-dissociation to the fluorescent compound 2,4 diamino-formylpteridine. In the 5-FU case the absorption spectra exhibit bands between 200 nm and 450 nm. The emission fluorescence spectra were measured between 400 nm and 600 nm, with λex = 350 nm for UV Hg lamp and with λex = 360 nm for laser irradiated samples; at irradiation with N2 laser emitted radiation the excitation spectra were measured in the range of 200 nm - 400 nm, with λem = 440 nm. New vascularity rapid destruction was observed for conjunctive impregnated with 5-FU solution whilst exposed to incoherent UV and visible light.

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

NASA Astrophysics Data System (ADS)

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

2018-05-01

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

Porphyrin synthesized from cashew nut shell liquid as part of a novel superparamagnetic fluorescence nanosystem

NASA Astrophysics Data System (ADS)

Clemente, C. S.; Ribeiro, V. G. P.; Sousa, J. E. A.; Maia, F. J. N.; Barreto, A. C. H.; Andrade, N. F.; Denardin, J. C.; Mele, G.; Carbone, L.; Mazzetto, S. E.; Fechine, P. B. A.

2013-06-01

Magnetic Fe3O4 nanoparticles with average size approximately 11 nm were first oleic acid coated to interact with the meso-porphyrin derivative from CNSL. This procedure produced a novel superparamagnetic fluorescent nanosystem (SFN) linked by van der Waals interactions. This system was characterized by transmission electron microscope, infrared spectroscopy, thermogravimetric analysis, magnetic measurements, UV-Vis absorption, and fluorescence emission measurements. These results showed that SFN has good thermal stability, excellent magnetization, and nanosized dimensions ( 13 nm). It exhibited emission peaks at 668 and 725 nm with a maximum emission at 467 nm of excitation wavelength. The type of interaction between porphyrin and magnetic nanoparticles allowed to obtain a material with interesting optical properties which might be used as an imaging agent for contrast in cells as well as heterogeneous photocatalysis.

Vibrio azureus emits blue-shifted light via an accessory blue fluorescent protein.

PubMed

Yoshizawa, Susumu; Karatani, Hajime; Wada, Minoru; Kogure, Kazuhiro

2012-04-01

Luminous marine bacteria usually emit bluish-green light with a peak emission wavelength (λ(max) ) at about 490 nm. Some species belonging to the genus Photobacterium are exceptions, producing an accessory blue fluorescent protein (lumazine protein: LumP) that causes a blue shift, from λ(max)  ≈ 490 to λ(max)  ≈ 476 nm. However, the incidence of blue-shifted light emission or the presence of accessory fluorescent proteins in bacteria of the genus Vibrio has never been reported. From our spectral analysis of light emitted by 16 luminous strains of the genus Vibrio, it was revealed that most strains of Vibrio azureus emit a blue-shifted light with a peak at approximately 472 nm, whereas other Vibrio strains emit light with a peak at around 482 nm. Therefore, we investigated the mechanism underlying this blue shift in V. azureus NBRC 104587(T) . Here, we describe the blue-shifted light emission spectra and the isolation of a blue fluorescent protein. Intracellular protein analyses showed that this strain had a blue fluorescent protein (that we termed VA-BFP), the fluorescent spectrum of which was almost identical to that of the in vivo light emission spectrum of the strain. This result strongly suggested that VA-BFP was responsible for the blue-shifted light emission of V. azureus. © 2012 Federation of European Microbiological Societies. Published by Blackwell Publishing Ltd. All rights reserved.

Laser induced fluorescence of dental caries

NASA Technical Reports Server (NTRS)

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.

LucY: A Versatile New Fluorescent Reporter Protein

PubMed Central

Auldridge, Michele E.; Franz, Laura P.; Bingman, Craig A.; Yennamalli, Ragothaman M.; Phillips, George N.; Mead, David; Steinmetz, Eric J.

2015-01-01

We report on the discovery, isolation, and use of a novel yellow fluorescent protein. Lucigen Yellow (LucY) binds one FAD molecule within its core, thus shielding it from water and maintaining its structure so that fluorescence is 10-fold higher than freely soluble FAD. LucY displays excitation and emission spectra characteristic of FAD, with 3 excitation peaks at 276nm, 377nm, and 460nm and a single emission peak at 530nm. These excitation and emission maxima provide the large Stokes shift beneficial to fluorescence experimentation. LucY belongs to the MurB family of UDP-N-acetylenolpyruvylglucosamine reductases. The high resolution crystal structure shows that in contrast to other structurally resolved MurB enzymes, LucY does not contain a potentially quenching aromatic residue near the FAD isoalloxazine ring, which may explain its increased fluorescence over related proteins. Using E. coli as a system in which to develop LucY as a reporter, we show that it is amenable to circular permutation and use as a reporter of protein-protein interaction. Fragmentation between its distinct domains renders LucY non-fluorescent, but fluorescence can be partially restored by fusion of the fragments to interacting protein domains. Thus, LucY may find application in Protein-fragment Complementation Assays for evaluating protein-protein interactions. PMID:25906065

LucY: A Versatile New Fluorescent Reporter Protein.

PubMed

Auldridge, Michele E; Cao, Hongnan; Sen, Saurabh; Franz, Laura P; Bingman, Craig A; Yennamalli, Ragothaman M; Phillips, George N; Mead, David; Steinmetz, Eric J

2015-01-01

We report on the discovery, isolation, and use of a novel yellow fluorescent protein. Lucigen Yellow (LucY) binds one FAD molecule within its core, thus shielding it from water and maintaining its structure so that fluorescence is 10-fold higher than freely soluble FAD. LucY displays excitation and emission spectra characteristic of FAD, with 3 excitation peaks at 276 nm, 377 nm, and 460 nm and a single emission peak at 530 nm. These excitation and emission maxima provide the large Stokes shift beneficial to fluorescence experimentation. LucY belongs to the MurB family of UDP-N-acetylenolpyruvylglucosamine reductases. The high resolution crystal structure shows that in contrast to other structurally resolved MurB enzymes, LucY does not contain a potentially quenching aromatic residue near the FAD isoalloxazine ring, which may explain its increased fluorescence over related proteins. Using E. coli as a system in which to develop LucY as a reporter, we show that it is amenable to circular permutation and use as a reporter of protein-protein interaction. Fragmentation between its distinct domains renders LucY non-fluorescent, but fluorescence can be partially restored by fusion of the fragments to interacting protein domains. Thus, LucY may find application in Protein-fragment Complementation Assays for evaluating protein-protein interactions.

LucY: A versatile new fluorescent reporter protein

DOE PAGES

Auldridge, Michele E.; Cao, Hongnan; Sen, Saurabh; ...

2015-04-23

We report on the discovery, isolation, and use of a novel yellow fluorescent protein. Lucigen Yellow (LucY) binds one FAD molecule within its core, thus shielding it from water and maintaining its structure so that fluorescence is 10-fold higher than freely soluble FAD. LucY displays excitation and emission spectra characteristic of FAD, with 3 excitation peaks at 276nm, 377nm, and 460nm and a single emission peak at 530nm. These excitation and emission maxima provide the large Stokes shift beneficial to fluorescence experimentation. LucY belongs to the MurB family of UDP-N-acetylenolpyruvylglucosamine reductases. The high resolution crystal structure shows that in contrastmore » to other structurally resolved MurB enzymes, LucY does not contain a potentially quenching aromatic residue near the FAD isoalloxazine ring, which may explain its increased fluorescence over related proteins. Using E. coli as a system in which to develop LucY as a reporter, we show that it is amenable to circular permutation and use as a reporter of protein-protein interaction. Fragmentation between its distinct domains renders LucY non-fluorescent, but fluorescence can be partially restored by fusion of the fragments to interacting protein domains. Thus, LucY may find application in Protein-fragment Complementation Assays for evaluating protein-protein interactions.« less

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

NASA Technical Reports Server (NTRS)

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

1994-01-01

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

Quantum sized Ag nanocluster assisted fluorescence enhancement in Tm{sup 3+}-Yb{sup 3+} doped optical fiber beyond plasmonics

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

Chattopadhyay, Rik; Haldar, Arindam; Paul, Mukul C.

2015-12-07

We report a process for enhancing fluorescence emission from conventional rare earth ions in optical fiber by metal nanocluster (MNC) in nonresonant indirect pumping. The process is completely different from formal metal enhanced fluorescence phenomenon as the MNCs are too small in size to support localized surface plasmon and the excitation wavelength is far from plasmon resonance frequency. We used an established theory of two coupled oscillators to explain the simultaneous enhancement of Ytterbium (Yb{sup 3+}) and Thulium (Tm{sup 3+}) emission by silver (Ag) NCs under nonresonant pumping in optical fiber. The fiber is pumped with a 980 nm fiber pigtailedmore » laser diode with input power of 20–100 mW to excite the Yb{sup 3+}. Four times enhancement of Yb{sup 3+} emission of 900–1100 nm and Tm{sup 3+} upconversion emission around 474 nm, 650 nm, and 790 nm is observed in the fiber with Ag NCs.« less

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

Harilal, Sivanandan S.; LaHaye, Nicole L.; Phillips, Mark C.

We use a two-dimensional laser-induced fluorescence spectroscopy technique to measure the coupled absorption and emission properties of atomic species in plasmas produced via laser ablation of solid aluminum targets at atmospheric pressure. Emission spectra from the Al I 394.4 nm and Al I 396.15 nm transitions are measured while a frequency-doubled, continuous-wave, Ti:Sapphire laser is tuned across the Al I 396.15 nm transition. The resulting two-dimensional spectra show the energy coupling between the two transitions via increased emission intensity for both transitions during resonant absorption of the continuous-wave laser at one transition. Time-delayed and gated detection of the emission spectrummore » is used to isolate the resonantly-excited fluorescence emission from the thermally-excited emission from the plasma. In addition, the tunable continuous-wave laser measures the absorption spectrum of the Al transition with ultra-high resolution after the plasma has cooled, resulting in narrower spectral linewidths than observed in emission spectra. Our results highlight that fluorescence spectroscopy employing continuous-wave laser re-excitation after pulsed laser ablation combines benefits of both traditional emission and absorption spectroscopic methods.« less

Foliar Reflectance and Fluorescence Responses for Corn and Soybean Plants Under Nitrogen Stress

NASA Technical Reports Server (NTRS)

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

2003-01-01

We are investigating the use of spectral indices derived from actively induced fluorescence spectra and passive optical spectra. We examined the influence of photosynthetic pigment, carbon (C) and nitrogen (N) content on the spectral fluorescence and passive optical property characteristics of mature, upper leaves from plants provided different N fertilizer application rates: 20%, 50%, 100% and 150% of recommended N levels. A suite of optical, fluorescence, and biophysical measurements were collected on leaves from field grown corn (Zea mays L.) and soybean plants (Glycine max L.) grown in pots (greenhouse + ambient sunlight. Steady state laser-induced fluorescence emission spectra (5 nm resolution) were obtained from adaxial and abaxial surfaces resulting from excitation at single wavelengths (280, 380 or 360, and 532 nm). For emission spectra produced by each of these excitation wavelengths, ratios of emission peaks were calculated, including the red far-red chlorophyll fluorescence (ChlF) ratio (F685/F740) and the far-red/green (F740/F525) ratio. High resolution (< 3 nm) optical spectra (350-2500 nm) of reflectance, transmittance, and absorptance were also acquired for both adaxial and abaxial leaf surfaces. Species differences were demonstrated for several optical parameters. A 'red edge' derivative ratio determined from transmittance spectra [as the maximum first deivative, between 650-750 nm, normalized to the value at 744 nm, or Dmax/D744], was strongly associated with the C/N ratio (r(exp 2) = 0.90, P +/- 0.001). This ratio, calculated from reflectance spectra, was inversely related to chlorophyll b content (r(exp 2) = 0.91, P +/- 0.001) as was the ChlF (F685/F740) ratio obtained with 532 nm excitation (r(exp 2) = 0.76, P +/- 0.01). Discrimination of N treatment groups was possible with specific fluorescence band ratios (e.g., F740/F525 obtained with 380 nm excitation). Higher ChlF and blue-green emissions were measured from the abaxial leaf surfaces. Abaxial surfaces also produced higher reflectances, in general, in the 400-800 nm spectrum.

A novel fluorescence "turn-on" sensor based on a photochromic diarylethene for the selective detection of Al(III)

NASA Astrophysics Data System (ADS)

Wang, Niansheng; Wang, Renjie; Tu, Yayi; Pu, Shouzhi; Liu, Gang

2018-05-01

A novel photochromic diarylethene with a triazole-containing 2-(2‧-phenoxymethyl)-benzothiazole group has been synthesized via "click" reaction. The diarylethene exhibited good photochromism and photoswitchable fluorescence. Its fluorescence emission intensity was enhanced 7-fold by acids, accompanied by the red-shift of emission peak from 526 nm to 566 nm and the concomitant color change from dark to bright flavogreen. The diarylethene selectively formed a 1:1 metal complex with Al3+, resulting in a "turn-on" fluorescence signal. The complexation - reaction between Al3+ and the diarylethene is reversible with the binding constant of 2.73 × 103 L mol-1. The limit of detection (LOD) of Al3+ was determined to be 5.94 × 10-8 mol L-1. Based on this unimolecular platform, a logic circuit was fabricated using the fluorescence emission intensity at 572 nm as the output and the combined stimuli of Al3+/EDTA and UV/Vis as the inputs.

Visualizing BPA by molecularly imprinted ratiometric fluorescence sensor based on dual emission nanoparticles.

PubMed

Lu, Hongzhi; Xu, Shoufang

2017-06-15

Construction of ratiometric fluorescent probe often involved in tedious multistep preparation or complicated coupling or chemical modification process. The emergence of dual emission fluorescent nanoparticles would simplify the construction process and avoids the tedious chemical coupling. Herein, we reported a facile strategy to prepare ratiometric fluorescence molecularly imprinted sensor based on dual emission nanoparticles (d-NPs) which comprised of carbon dots and gold nanoclusters for detection of Bisphenol A (BPA). D-NPs emission at 460nm and 580nm were first prepared by seed growth co-microwave method using gold nanoparticles as seeds and glucose as precursor for carbon dots. When they were applied to propose ratiometric fluorescence molecularly imprinted sensor, the preparation process was simplified, and the sensitivity of sensor was improved with detection limit of 29nM, and visualizing BPA was feasible based on the distinguish fluorescence color change. The feasibility of the developed method in real samples was successfully evaluated through the analysis of BPA in water samples with satisfactory recoveries of 95.9-98.9% and recoveries ranging from 92.6% to 98.6% in canned food samples. When detection BPA in positive feeding bottles, the results agree well with those obtained by accredited method. The developed method proposed in this work to prepare ratiometric fluorescence molecularly imprinted sensor based on dual emission nanoparticles proved to be a convenient, reliable and practical way to prepared high sensitive and selective fluorescence sensors. Copyright © 2017 Elsevier B.V. All rights reserved.

Synchronous fluorescence spectroscopy for analysis of wine and wine distillates

NASA Astrophysics Data System (ADS)

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

2015-01-01

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

Dendritic copper phthalocyanine with aggregation induced blue emission and solid-state fluorescence

NASA Astrophysics Data System (ADS)

Wang, Jiayi; Pan, Lin; Zhou, Xuefei; Jia, Kun; Liu, Xiaobo

2016-09-01

In this work, dendritic copper phthalocyanine (CuPc) showing obvious aggregation induced emission (AIE) and strong solid-state fluorescence was synthesized. It was found that synthesized CuPc can be easily solubilized in polar aprotic solvent, where no fluorescence signal was detected. Interestingly, both the CuPc aggregates in solution and solid-state powder exhibited strong fluorescence emission around 480 nm, which should be attributed to the restriction of intramolecular rotation as rationalized in aggregation induced emission framework. Meanwhile the obvious crystalline enhanced solid-state fluorescent emission is observed for CuPc powder.

Chlorophyll Proteins of Photosystem I 1

PubMed Central

Mullet, John E.; Burke, John J.; Arntzen, Charles J.

1980-01-01

Data are presented which suggest the existence of a light-harvesting pigment-protein complex which is functionally and structurally associated with photosystem I (PSI) reaction centers. These observations are based on techniques which allow isolation of PSI using minimal concentrations of Triton X-100. Properties of density and self aggregation allowed purification of a “native” PSI complex. The isolated PSI particles appear as 106 Å spherical subunits when viewed by freeze fracture microscopy. When incorporated into phosphatidyl choline vesicles, the particles lose self-aggregation properties and disperse uniformly within the lipid membrane. The isolated PSI preparation contains 100 ± 10 chlorophylls/P700 (Chl a/b ratio greater than 18); this represents a recovery of 27% of the original chloroplast membrane Chl. These particles were enriched in Chl a forms absorbing at 701 to 710 nm. Chl fluorescence at room temperature exhibited a maximum at 690 nm with a pronounced shoulder at 710 nm. At 77 K, peak fluorescence emission was at 736 nm; in the presence of dithionite an additional fluorescence maximum at 695 nm was obtained at 77 K. This dual fluorescence emission peak for the PSI particles is evidence for at least two Chl populations within the PSI membrane subunit. The fluorescence emission observed at 695 nm was identified as arising from the core of PSI which contains 40 Chl/P700 (PSI-40). This core complex, derived from native PSI particles, was enriched in Chl a absorbing at 680 and 690 nm and fluorescing with maximal emission at 694 nm at 77 K. PSI particles consisting of the PSI core complex plus 20 to 25 Chl antennae (65 Chl/P700) could also be derived from native PSI complexes. These preparations were enriched in Chl a forms absorbing at 697 nm and exhibited a 77 K fluorescence emission maximum at 722 nm. A comparison of native PSI particles which contain 110 Chl/P700 (PSI-110) and PSI particles containing 65 Chl/P700 (PSI-65) provides evidence for the existence of a peripheral Chl-protein complex tightly associated in the native PSI complex. The native PSI subunits contain polypeptides of 22,500 to 24,500 daltons which are not found in the PSI-65 or PSI-40 subfractions. It is suggested that these polypeptides function to bind 40 to 45 Chl per structural complex, including the Chl which emits fluorescence at 736 nm. A model for the organization of Chl forms is presented in which the native PSI membrane subunit consists of a reaction center core complex plus two regions of associated light-harvesting antennae. The presence of energy “sinks” within the antennae is discussed. Images PMID:16661288

Studies of the DOM aqueous extracts from coastal marine sediments

NASA Astrophysics Data System (ADS)

Sakellariadou, F.

2012-04-01

Dissolved organic matter (DOM) represents a major exchangeable organic pool playing an outstanding role in the ocean carbon cycle. It has a complex chemical structure made up of a wide range of organic molecules. The composition of DOM depends on the sources proximity and the exposure to any sort of degradation mechanism. The coloured (or chromophoric) dissolved organic matter (CDOM), representing the optically active fraction of DOM, consists of aromatic rings able to absorb light in the visible and UV regions (Kirk, 1994) and fluorophoric molecules that emit light. The main fluorophoric moieties of CDOM are humic material with a blue fluorescence and protein material with an ultraviolet (UV) fluorescence (Mopper and Schultz, 1993). Dissolved organic matter interacts with pollutants either by enhancing their bioavailability or by influencing their transportation to the soluble phase. In addition, DOM affects the remineralisation of carbon and its preservation in marine sediments. Referring to its origin, it can be terrestrial, freshwater or marine one. Fluorescence spectroscopy is a technique widely applied for the identification and characterization of organic matter, being fast, simple, non-destructive and sensitive. In addition, the fluorescence analysis for the physico-chemical characterization of organic matter requires a small amount of aqueous sample at a low concentration, in comparison with the large sample volumes needed for conventional techniques. At the present study coastal sediment samples were collected from Messiniakos gulf in the south western Peloponnese in South Greece. Messiniakos gulf has a seabed dominated by very abrupt inclinations reaching depths of more than 1000m. All samples, according to their grain size, are classified as fine clayey silt. Dissolved organic matter was extracted under gentle extraction conditions (4 mM CaCl2 solution). The various classes of organic components present at the DOM aqueous extracts were characterised by fluorescence spectroscopy technique as DOM fluorescence is a property furnishing valuable information for its composition and biogeochemical cycling. Fluorescence spectra were recorded using a Perkin-Elmer LS 55 luminescence spectrophotometer equipped with the WinLab 4.00.02 software for data processing. Conventional mono-dimensional emission, excitation and synchronous-scan excitation spectra were recorded. Mono dimensional emission spectra were recorded over the range 380-600 nm at a constant excitation wavelength of 360 nm. Excitation spectra were recorded over the range 300-500 nm at a fixed emission wavelength of 520 nm. Synchronous-scan excitation spectra were measured by scanning simultaneously both the excitation and the emission wavelengths (from 300 to 550 nm), while maintaining a constant, optimised wavelength difference Δλ (λem - λexc) = 18 nm. (Senesi et al., 1991). Total Luminescence Spectra (3D fluorescence spectra) were obtained in the form of excitation/emission matrix spectra (or contour maps) by scanning the wavelength emission over the range 300 to 600 nm, while the excitation wavelength was increased sequentially by 5-nm steps from 250 to 500 nm. In comparison with the limited provided information from the conventional fluorescence spectroscopy, EEM analysis seems to be sufficiently sensitive to distinguish between the various types of marine gelbstoff as well as to help estimating the contribution of each of them. Humification indices according to Ohno (2002) and Zsolnay (1999) were calculated. The Humification index (HIX) according to Ohno is calculated by dividing the emission intensity in the 435 to 480 nm region by the emission intensity in the 300 to 345 nm region; HIX quantifies the red shift of the emission spectra toward longer wavelengths with increasing humification. The HI index according to Zsolnay is defined as the area in the upper quarter (Σ435-480nm) of the usable emission peak divided by the area in the lower usable quarter (Σ300-445nm). All fluorescence spectra were thoroughly evaluated for the classification of chromophoric units present. Variations were observed according to the sampling site and more precisely its proximity to the coastline and the corresponding water column's depth; the oceanographic characteristics allowing or obstructing sea water circulation; as well as the proximity of each sample to the seabed, in other words the characterization of each sediment sample as surface of subsurface one.

Study on fluorescence spectra of thiamine, riboflavin and pyridoxine

NASA Astrophysics Data System (ADS)

Yang, Hui; Xiao, Xue; Zhao, Xuesong; Hu, Lan; Lv, Caofang; Yin, Zhangkun

2016-01-01

This paper presents the intrinsic fluorescence characteristics of vitamin B1, B2 and B6 measured with 3D fluorescence Spectrophotometer. Three strong fluorescence areas of vitamin B2 locate at λex/λem=270/525nm, 370/525nm and 450/525nm, one fluorescence areas of vitamin B1 locates at λex/λem=370/460nm, two fluorescence areas of vitamin B6 locate at λex/λem=250/370nm and 325/370nm were found. The influence of pH of solution to the fluorescence profile was also discussed. Using the PARAFAC algorithm, 10 vitamin B1, B2 and B6 mixed solutions were successfully decomposed, and the emission profiles, excitation profiles, central wavelengths and the concentration of the three components were retrieved precisely through about 5 iteration times.

A fluorescence approach to the unfolding thermodynamics of horseradish peroxidase based on heme degradation by hydrogen peroxide

NASA Astrophysics Data System (ADS)

Ke, Zhigang; Ma, Shanshan; Li, Lamei; Huang, Qing

2016-07-01

Horseradish peroxidase (HRP) is a classical heme-containing protein which has been applied in many fields. The prosthetic group heme in HRP, especially in unfolded state, can react with hydrogen peroxide (H2O2) to produce a fluorescent product with the maximum emission wavelength at 450 nm. Utilizing this emission band as a fluorescence probe, the unfolding process of HRP in urea can be assessed quantitatively, and the calculated thermodynamic parameters are consistent with those determined by circular dichroism (CD) at 222 nm and steady-state tryptophan (Trp) fluorescence methods.

Leaf Level Chlorophyll Fluorescence Emission Spectra: Narrow Band versus Full 650-800 nm Retrievals

NASA Astrophysics Data System (ADS)

Middleton, E.; Zhang, Q.; Campbell, P. K.; Huemmrich, K. F.; Corp, L.; Cheng, Y.

2012-12-01

Recently, chlorophyll fluorescence (ChlF) retrievals in narrow spectral regions (< 1 nm, between 750-770 nm) of the near infrared (NIR) region of Earth's reflected radiation have been achieved from satellites, including the Japanese GOSAT and the European Space Agency's Sciamachy/Envisat. However, these retrievals sample the total full-spectrum ChlF and are made at non-optimal wavelengths since they are not located at the peak fluorescence emission features. We wish to estimate the total full-spectrum ChlF based on emissions obtained at selected wavelengths. For this, we drew upon leaf emission spectra measured on corn leaves obtained from a USDA experimental cornfield in MD (USA). These emission spectra were determined for the adaxial and abaxial (i.e., top and underside) surfaces of leaves measured throughout the 2008 and 2011 growing seasons (n>400) using a laboratory instrument (Fluorolog-3, Horiba Scientific, USA), recorded in either 1 nm or 5 nm increments with monochromatic excitation wavelengths of either 532 or 420 nm. The total ChlF signal was computed as the area under the continuous spectral emission curves, summing the emission intensities (counts per second) per waveband. The individual narrow (1 or 5 nm) waveband emission intensities were linearly related to full emission values, with variable success across the spectrum. Equations were developed to estimate total ChlF from these individual wavebands. Here, we report the results for the average adaxial/abaxial emissions. Very strong relationships were achieved for the relatively high fluorescence intensities at the red chlorophyll peak, centered at 685 nm (r2= 0.98, RMSE = 5.53 x 107 photons/s) and in the nearby O2-B atmospheric absorption feature centered at 688 nm (r2 = 0.94, RMSE = 4.04 x 107), as well as in the far-red peak centered at 740 nm (r2=0.94, RMSE = 5.98 x107). Very good retrieval success occurred for the O2-A atmospheric absorption feature on the declining NIR shoulder centered at 760 nm (r2 = 0.88, RMSE = 7.54 x 107). When perfect retrievals were assumed (0% noise), retrievals remained good in the low emission regions on either side of the peaks-- those associated with the H alpha line at 655 nm (r2 = 0.83, RMSE =8.87 x 107) and the far-NIR wavelengths recently utilized for satellite retrievals: a K line at 770 nm (r2 = 0.85, RMSE = 8.36 x 107) and the 750-770 nm interval (r2 = 0.88, RMSE = 6.92 x 107). However, the atmosphere and satellite observations are expected to add noise to retrievals. Adding 5% random error to these relationships did not seriously impair the retrieval successes in the red and far-red peaks (r2 ~ 0.85, RMSEs = 6.31 x 107). A greater impact occurred (reducing retrieval success by ~10%) when adding 5% noise for the far-NIR narrow band at 770 nm (r2 ~ 0.70, RMSE ~ 8.5 x 107). When a 10% random error was added, the retrieval successes fell to ~68 ± 7% for all retrieval wavebands, and RMSEs increased by a factor of 10. This laboratory approach will be critical to calibrate space borne retrievals, but additional information across plant species is needed. Furthermore, this experiment indicates that ChlF retrievals from space should include information from the red and far-red peak emission regions, since the true total fluorescence signal is the desired parameter for Earth carbon and energy budgets.

Composite fluorescent nanoparticles for biomedical imaging.

PubMed

Pansare, Vikram J; Bruzek, Matthew J; Adamson, Douglas H; Anthony, John; Prud'homme, Robert K

2014-04-01

In the rapidly expanding field of biomedical imaging, there is a need for nontoxic, photostable, and nonquenching fluorophores for fluorescent imaging. We have successfully encapsulated a new, extremely hydrophobic, pentacene-based fluorescent dye within polymeric nanoparticles (NPs) or nanocarriers (NCs) via the Flash NanoPrecipitation (FNP) process. Nanoparticles and dye-loaded micelles were formulated by FNP and characterized by dynamic light scattering, fluorescence spectroscopy, UV-VIS absorbance spectroscopy, and confocal microscopy. These fluorescent particles were loaded from less than 1% to 78% by weight core loading and the fluorescence maximum was found to be at 2.3 wt.%. The particles were also stably formed at 2.3% core loading from 20 up to 250 nm in diameter with per-particle fluorescence scaling linearly with the NC core volume. The major absorption peaks are at 458, 575, and 625 nm, and the major emission peaks at 635 and 695 nm. In solution, the Et-TP5 dye displays a strong concentration-dependent ratio of the emission intensities of the first two emission peaks, whereas in the nanoparticle core the spectrum is independent of concentration over the entire concentration range. A model of the fluorescence quenching was consistent with Förster resonant energy transfer as the cause of the quenching observed for Et-TP5. The Förster radius calculated from the absorption and emission spectra of Et-TP5 is 4.1 nm, whereas the average dye spacing in the particles at the maximum fluorescence is 3.9 nm. We have successfully encapsulated Et-TP5, a pentacene derivative dye previously only used in light-emitting diode applications, within NCs via the FNP process. The extreme hydrophobicity of the dye keeps it encapsulated in the NC core, its extended pentacene structure gives it relatively long wavelength emission at 695 nm, and the pentacene structure, without oxygen or nitrogen atoms in its core, makes it highly resistant to photobleaching. Its bulky side groups minimize self-quenching and localization within the nanoparticle core prevents interaction of the dye with biological surfaces, or molecules in diagnostic assays. Loading of dye in the NP core allows 25 times more dye to be delivered than if it were conjugated onto the nanocarrier surface. The utility of the dye for quantifying nanoparticle binding is demonstrated. Studies to extend the wavelength range of these pentacene dyes into the near infra-red are underway.

One-pot and ultrafast synthesis of nitrogen and phosphorus co-doped carbon dots possessing bright dual wavelength fluorescence emission

NASA Astrophysics Data System (ADS)

Sun, Xiangcheng; Brückner, Christian; Lei, Yu

2015-10-01

Very brief microwave heating of aniline, ethylene diamine, and phosphoric acid in water at ambient pressure generated nitrogen and phosphorus co-doped carbon dots (N,P-CDs) that exhibit bright dual blue (centred at 450 nm; 51% quantum yield) and green (centred at 510 nm, 38% quantum yield) fluorescence emission bands. The N,P-CDs were characterized using TEM, XRD, XPS, IR, UV-vis, and fluorescence spectroscopy, demonstrating their partially crystalline carbon, partially amorphous structures, and the incorporation of O, N, and P into the carbogenic scaffold. The N,P-CDs demonstrated excitation-dependent and nearly pH-independent emission properties. The unique dual emission properties lay the foundation for the use of N,P-CDs in ratiometric sensing applications.Very brief microwave heating of aniline, ethylene diamine, and phosphoric acid in water at ambient pressure generated nitrogen and phosphorus co-doped carbon dots (N,P-CDs) that exhibit bright dual blue (centred at 450 nm; 51% quantum yield) and green (centred at 510 nm, 38% quantum yield) fluorescence emission bands. The N,P-CDs were characterized using TEM, XRD, XPS, IR, UV-vis, and fluorescence spectroscopy, demonstrating their partially crystalline carbon, partially amorphous structures, and the incorporation of O, N, and P into the carbogenic scaffold. The N,P-CDs demonstrated excitation-dependent and nearly pH-independent emission properties. The unique dual emission properties lay the foundation for the use of N,P-CDs in ratiometric sensing applications. Electronic supplementary information (ESI) available: Detailed experimental section, XRD, FTIR, explosive sensing and the applications results. See DOI: 10.1039/c5nr05549k

A Preliminary Study of the Effects of pH upon Fluorescence in Suspensions of Prevotella intermedia.

PubMed

Hope, Christopher K; Billingsley, Karen; de Josselin de Jong, Elbert; Higham, Susan M

2016-01-01

The quantification of fluorescence in dental plaque is currently being developed as a diagnostic tool to help inform and improve oral health. The oral anaerobe Prevotella intermedia exhibits red fluorescence due to the accumulation of porphyrins. pH affects the fluorescence of abiotic preparations of porphyrins caused by changes in speciation between monomers, higher aggregates and dimers, but this phenomenon has not been demonstrated in bacteria. Fluorescence spectra were obtained from suspensions of P. intermedia that were adjusted to pHs commensurate with the range found within dental plaque. Two fluorescent motifs were identified; 410 nm excitation / 634 nm emission (peak A) and 398 nm excitation / 622 nm emission (peak B). A transition in the fluorescence spectra was observed from peak A to peak B with increasing pH which was also evident as culture age increased from 24 hours to 96 hours. In addition to these 'blue-shifts', the intensity of peak A increased with pH whilst decreasing with culture age from 24 to 96 hours. A bacterium's relationship with the local physiochemical environment at the time of image capture may therefore affect the quantification of dental plaque fluorescence.

Interpretation of the fluorescence signatures from vegetation

NASA Astrophysics Data System (ADS)

Buschmann, C.

Vegetation emits fluorescence as part of the energy taken up by absorption %of solar radiation from UV to the visible. This fluorescence consists of light with low intensity (only few percents of the reflected light) emitted from the leaves. The fluorescence emission of a green leaf is characterized by four bands with maxima in the blue (440 nm), green (520 nm), red (690 nm) and far red (740 nm) spectral region. The intensity of fluorescence in the maxima of the emission spectrum varies depending on the following six basic parameters which must be taken into account for the interpretation of fluorescence signatures from vegetation: (a) content of the fluorophores (ferulic acid, chlorophyll a), (b) temperature of the leaf, (c) penetration of excitation light into the leaf, (d) emission of fluorescence from the leaf (re-absorption inside the leaf tissue), (e) photosynthetic activity of the leaf, (f) non-radiative decay (heat production) parallel to the fluorescence The ratios between the intensities of the maxima (F440/F690, F440/F520, F690/F740) are used as characteristic fluorescence parameter. The wide range of changes of these ratios caused by differences in the leaf tissue (aerial interspaces, variegated/homogeneous green leaves), various types of stress (UV, photoinhibition, sun exposure, heat, water deficiency, N-deficiency) and chemicals (inhibitors, fertilizers) can be explained by changes of the six basic parameters. It will be shown that the interpretation of the fluorescence signatures, in most cases, must be based on a complex consideration of more than one of the basic parameters.

Investigations on the effects of the Stark splitting on the fluorescence behaviors in Yb3+-doped silicate, tellurite, germanate, and phosphate glasses

NASA Astrophysics Data System (ADS)

Zhang, Liaolin; Xia, Yu; Shen, Xiao; Yang, Runlan; Wei, Wei

2018-01-01

In this work, we systematically studied the spectroscopic characteristics of Yb3+ doped germanate, phosphate, silicate, and tellurite glasses. The emission peak beyond 976 nm showed irregular shift from 1001 nm to 1023 nm when Yb3+ in different glass matrices. It was associated with the Stark splitting of 2F7/2 and the emission intensities ratio between the transition from the lowest Stark splitting energy level of 2F5/2 to the Stark splitting energy levels of 2F7/2, e to b and that of e to d. Larger Stark splitting of 2F7/2 results in the red-shift of the near infrared emission band at room temperature and larger ratio results in the blue-shift of emission band. The fluorescence lifetimes of Yb3+ doped germanate, phosphate, silicate, and tellurite glasses were measured to be 0.94, 0.82, 1.51, and 0.66 ms, respectively. The fluorescence lifetime was associated with the reabsorption of Yb3+, which larger absorption cross section at the emission band results in larger reabsorption, then leads to the shorter near infrared fluorescence lifetime.

Single particle size and fluorescence spectra from emissions of burning materials in a tube furnace to simulate burn pits

NASA Astrophysics Data System (ADS)

Pan, Yong-Le; Houck, Joshua D. T.; Clark, Pamela A.; Pinnick, Ronald G.

2013-08-01

A single-particle fluorescence spectrometer (SPFS) and an aerodynamic particle sizer were used to measure the fluorescence spectra and particle size distribution from the particulate emissions of 12 different burning materials in a tube furnace to simulate open-air burning of garbage. Although the particulate emissions are likely dominated by particles <1 μm diameter, only the spectra of supermicron particles were measured here. The overall fluorescence spectral profiles exhibit either one or two broad bands peaked around 300-450 nm within the 280-650 nm spectral range, when the particles are illuminated with a 263-nm laser. Different burning materials have different profiles, some of them (cigarette, hair, uniform, paper, and plastics) show small changes during the burning process, and while others (beef, bread, carrot, Styrofoam, and wood) show big variations, which initially exhibit a single UV peak (around 310-340 nm) and a long shoulder in visible, and then gradually evolve into a bimodal spectrum with another visible peak (around 430-450 nm) having increasing intensity during the burning process. These spectral profiles could mainly derive from polycyclic aromatic hydrocarbons with the combinations of tyrosine-like, tryptophan-like, and other humic-like substances. About 68 % of these single-particle fluorescence spectra can be grouped into 10 clustered spectral templates that are derived from the spectra of millions of atmospheric aerosol particles observed in three locations; while the others, particularly these bimodal spectra, do not fall into any of the 10 templates. Therefore, the spectra from particulate emissions of burning materials can be easily discriminated from that of common atmospheric aerosol particles. The SFFS technology could be a good tool for monitoring burning pit emissions and possibly for distinguishing them from atmospheric aerosol particles.

The spectral properties of (-)-epigallocatechin 3-O-gallate (EGCG) fluorescence in different solvents: dependence on solvent polarity.

PubMed

Snitsarev, Vladislav; Young, Michael N; Miller, Ross M S; Rotella, David P

2013-01-01

(-)-Epigallocatechin 3-O-gallate (EGCG) a molecule found in green tea and known for a plethora of bioactive properties is an inhibitor of heat shock protein 90 (HSP90), a protein of interest as a target for cancer and neuroprotection. Determination of the spectral properties of EGCG fluorescence in environments similar to those of binding sites found in proteins provides an important tool to directly study protein-EGCG interactions. The goal of this study is to examine the spectral properties of EGCG fluorescence in an aqueous buffer (AB) at pH=7.0, acetonitrile (AN) (a polar aprotic solvent), dimethylsulfoxide (DMSO) (a polar aprotic solvent), and ethanol (EtOH) (a polar protic solvent). We demonstrate that EGCG is a highly fluorescent molecule when excited at approximately 275 nm with emission maxima between 350 and 400 nm depending on solvent. Another smaller excitation peak was found when EGCG is excited at approximately 235 nm with maximum emission between 340 and 400 nm. We found that the fluorescence intensity (FI) of EGCG in AB at pH=7.0 is significantly quenched, and that it is about 85 times higher in an aprotic solvent DMSO. The Stokes shifts of EGCG fluorescence were determined by solvent polarity. In addition, while the emission maxima of EGCG fluorescence in AB, DMSO, and EtOH follow the Lippert-Mataga equation, its fluorescence in AN points to non-specific solvent effects on EGCG fluorescence. We conclude that significant solvent-dependent changes in both fluorescence intensity and fluorescence emission shifts can be effectively used to distinguish EGCG in aqueous solutions from EGCG in environments of different polarity, and, thus, can be used to study specific EGCG binding to protein binding sites where the environment is often different from aqueous in terms of polarity.

Modification of fluorescence and optical properties of Rhodamine B dye doped PVA/Chitosan polymer blend films

NASA Astrophysics Data System (ADS)

Padmakumari, R.; Ravindrachary, V.; Mahantesha, B. K.; Sagar, Rohan N.; Sahanakumari, R.; Bhajantri, R. F.

2018-05-01

Pure and Rhodamine B doped Poly (vinyl alcohol)/Chitosan composite films are prepared using solution casting method. Fourier transforms infrared spectra (FTIR), Ultraviolet-Visible (UV-Vis), fluorescence studies were used to characterize the prepared polymer films. The FT-IR results show that the appearance of new peaks along with shift in peak positions indicates the interaction of Rhodamine B with PVA-CS blend. Optical absorption edge, band gap and activation energy were determined from UV-Visible studies. The optical absorption edge increases, band gap decreases and activation energy increases with dopant concentration respectively. The corresponding emission spectra were studied using fluorescence spectroscopy. From the fluorescence study the quenching phenomena are observed in emission wavelength range of 607nm-613nm upon excitation with absorption maxima 443nm.

Intrinsic fluorescence spectra characteristics of vitamin B1, B2, and B6

NASA Astrophysics Data System (ADS)

Yang, Hui; Xiao, Xue; Zhao, Xuesong; Hu, Lan; Lv, Caofang; Yin, Zhangkun

2015-11-01

This paper presents the intrinsic fluorescence characteristics of vitamin B1, B2 and B6 measured with 3D fluorescence Spectrophotometer. Three strong fluorescence areas of vitamin B2 locate at λex/λem=270/525nm, 370/525nm and 450/525nm, one fluorescence areas of vitamin B1 locates at λex/λem=370/460nm, two fluorescence areas of vitamin B6 locates at λex/λem=250/370nm and 325/370nm were found. The influence of pH of solution to the fluorescence profile was also discussed. Using the PARAFAC algorithm, 10 vitamin B1, B2 and B6 mixed solutions were successfully decomposed, and the emission profiles, excitation profiles, central wavelengths and the concentration of the three components were retrieved precisely through about 5 iteration times.

The exploration of the characteristics of the hyperglycemia serum fluorescence spectrum

NASA Astrophysics Data System (ADS)

Wang, Lexin; Zhao, Zhimin; Chen, Hui; Li, Peng; Xin, Yujun

2008-12-01

Now, spectra technology is widely used in the biomedicine research,so this study investigates variation of the fluorescence spectra in different excitation wavelength, and the spectra of serum with different glucose concentration is tested in the excitation wavelength of 240nm to 280nm. The experimental result shows that the correlation between the serum fluorescence intensity and the excitation light is very close, when the excitation light is in the ultraviolet wave band, the fluorescence of serum is intensive. There is a violent fluorescence emission wavelength, which is 300nm to 410nm, while the excitation wavelength ranges from 220nm to 290nm, and the peaks wavelength are 330nm and 370nm. From 240nm to 280nm, the serum fluorescence intensity increases synchronously with the glucose concentration, and the relationship between the fluorescence peak wavelength and the glucose concentration is almost in line. In this way the blood sugar concentration can be estimated by the fluorescence spectra peak wavelength when the excitation wavelength is from 240nm to 280nm, which is effective. It provides experimental foundation for the wide use of spectra technology in medical diagnose, and the effectiv method to test the blood sugar concentration.

Ratiometric fluorescent detection of chromium(VI) in real samples based on dual emissive carbon dots.

PubMed

Ma, Yunxia; Chen, Yonglei; Liu, Juanjuan; Han, Yangxia; Ma, Sudai; Chen, Xingguo

2018-08-01

As we know, hexavalent chromium (Cr(VI)) was usually used as an additive to improve the color fastness during the printing and dyeing process, and thus posing tremendous threat to our health and living quality. In this work, the dual emissive carbon dots (DECDs) were synthesized through hydrothermal treatment of m-aminophenol and oxalic acid. The obtained DECDs not only exhibited dual emission fluorescence peaks (430 nm, 510 nm) under the single excitation wavelength of 380 nm, but also possessed good water solubility and excellent fluorescence stability. A ratiometric fluorescent method for the determination of Cr(VI) was developed using the DECDs as a probe. Under the optimal conditions, a linear range was obtained from 2 to 300 μM with a limit of detection of 0.4 μM. Furthermore, the proposed ratiometric fluorescent method was applied to the analysis of Cr(VI) in textile, steel, industrial wastewater and chromium residue samples with satisfactory recoveries (88.4-106.8%). Copyright © 2018 Elsevier B.V. All rights reserved.

A novel fluorescence "turn-on" sensor based on a photochromic diarylethene for the selective detection of Al(III).

PubMed

Wang, Niansheng; Wang, Renjie; Tu, Yayi; Pu, Shouzhi; Liu, Gang

2018-05-05

A novel photochromic diarylethene with a triazole-containing 2-(2'-phenoxymethyl)-benzothiazole group has been synthesized via "click" reaction. The diarylethene exhibited good photochromism and photoswitchable fluorescence. Its fluorescence emission intensity was enhanced 7-fold by acids, accompanied by the red-shift of emission peak from 526nm to 566nm and the concomitant color change from dark to bright flavogreen. The diarylethene selectively formed a 1:1 metal complex with Al 3+ , resulting in a "turn-on" fluorescence signal. The complexation - reaction between Al 3+ and the diarylethene is reversible with the binding constant of 2.73×10 3 Lmol -1 . The limit of detection (LOD) of Al 3+ was determined to be 5.94×10 -8 molL -1 . Based on this unimolecular platform, a logic circuit was fabricated using the fluorescence emission intensity at 572nm as the output and the combined stimuli of Al 3+ /EDTA and UV/Vis as the inputs. Copyright © 2018 Elsevier B.V. All rights reserved.

Different sensing modes of fluoride and acetate based on a calix[4]arene with 25,27-bistriazolylmethylpyrenylacetamides.

PubMed

Hung, Hao-Chih; Chang, Yung-Yu; Luo, Liyang; Hung, Chen-Hsiung; Diau, Eric Wei-Guang; Chung, Wen-Sheng

2014-02-01

25,27-Bis{1'-N-(1-pyrenyl)-aminocarbonylmethyl-1H-[1',2',3']tri-azolyl-4'-methoxy}-26,28-dihydroxycalix[4]arene, 4, is synthesized as a fluorescent chemosensor for the selective detection of both anions and ion pairs in MeCN. Sensor 4 uses bis-triazoles as ligands to bind a metal ion, bis-amides and bis-triazoles as the sites to recognize anions, and pyrenes as fluorophores. Among eight anions screened, chemosensor 4 showed a marked fluorescence change toward F(-), H2PO4(-) and AcO(-), but 4 responded to each anion in a distinct way. In the presence of F(-) at low concentrations, the dynamic excimer emission of compound 4 at λ(max) 482 nm was quenched, but an emission at λ(max) 472 nm appeared at large doses of F(-). A control compound 6 showed very similar red shifts in the UV-vis and excitation spectra as 4 did, and its 472 nm emission band grew as the fluoride doses increased. Thus, the growth of the 472 nm emission of 4 and 6 in the presence of excess F(-) may be because strong H-bonding interactions of amido protons with F(-) favoured the formation of pyrene dimers in the ground state with charge transfer characteristics. The addition of H2PO4(-), unlike F(-), to a solution of 4 showed an enhanced monomer emission but a decreased excimer emission (λ(max) 482 nm). Adding AcO(-) to 4 produced a systematic change from a dynamic excimer (λ(max) 482 nm) to λ(max) 472 nm but with very little change in the UV-vis spectrum. Time-resolved fluorescence measurements on compound 6 with F(-) and AcO(-) confirmed that the 472 nm emission band mainly came from static excimers for the former, but was partly from a dynamic excimer for the latter because it contained a growth component in the fluorescence decay traces. Without pre-treatment with an anion, chemosensor 4 showed recognition of only metal ions Cu(2+), Hg(2+) and Cr(3+), but it became sensitive to Ag(+) when it was pretreated with fluoride.

Hydrophobic-carbon-dot-based dual-emission micelle for ratiometric fluorescence biosensing and imaging of Cu2+ in liver cells.

PubMed

Lu, Linlin; Feng, Chongchong; Xu, Jie; Wang, Fengyang; Yu, Haijun; Xu, Zhiai; Zhang, Wen

2017-06-15

Copper is closely related to liver damage, therefore, it is essential to develop a simple and sensitive strategy to detect copper ions (Cu 2+ ) in liver cells. A hydrophobic carbon dots (HCDs)-based dual-emission fluorescent probe for Cu 2+ was prepared by encapsulating HCDs in micelles formed by self-assembly of amphiphilic polymer DSPE-PEG and tetrakis (4-carboxyphenyl) porphyrin (TCPP)-modified DSPE-PEG. The obtained probe showed characteristic fluorescence emissions of HCDs and TCPP with large emission shift of 170nm with single-wavelength excitation. In the presence of Cu 2+ , the fluorescence of TCPP was quenched and that of HCDs remained unchanged, displaying ratiometric fluorescence response to Cu 2+ . The developed probe exhibited high sensitivity (detection limit down to 36nM) and selectivity to Cu 2+ over other substances, and the probe was used to image the changes of Cu 2+ level in liver cells successfully. Copyright © 2017 Elsevier B.V. All rights reserved.

Optical Properties of CdSe/ZnS Nanocrystals

PubMed Central

Gaigalas, Adolfas K; DeRose, Paul; Wang, Lili; Zhang, Yu-Zhong

2014-01-01

Measurements are presented of the absorbance, fluorescence emission, fluorescence quantum yield, and fluorescence lifetime of CdSe/ZnS nanocrystals, also known as quantum dots (QDs). The study included three groups of nanocrystals whose surfaces were either passivated with organic molecules, modified further with carboxyl groups, or conjugated with CD14 mouse anti-human antibodies. The surface modifications had observable effects on the optical properties of the nanocrystals. The oscillator strength (OS) of the band edge transition was about 1.0 for the nanocrystals emitting at 565 nm, 605 nm, and 655 nm. The OS could not be determined for QDs with emission at 700 nm and 800 nm. The fluorescence lifetimes varied from 26 ns for nanocrystals emitting near 600 nm to 150 ns for nanocrystals emitting near 800 nm. The quantum yield ranged between 0.4 and 0.9 for the nanocrystals in this study. A brightness index (BI) was used to evaluate the suitability of the nanocrystal labels for flow cytometer measurements. Most QD labels are at least as bright as fluorescein for applications in flow cytometer assays with 488 nm excitation. For optimal brightness the QDs should be excited with 405 nm light. We observed a strong dependence of the QD absorbance at 250 nm on the surface modification of the QD. PMID:26601047

Red fluorescent protein responsible for pigmentation in trematode-infected Porites compressa tissues.

PubMed

Palmer, Caroline V; Roth, Melissa S; Gates, Ruth D

2009-02-01

Reports of coral disease have increased dramatically over the last decade; however, the biological mechanisms that corals utilize to limit infection and resist disease remain poorly understood. Compromised coral tissues often display non-normal pigmentation that potentially represents an inflammation-like response, although these pigments remain uncharacterized. Using spectral emission analysis and cryo-histological and electrophoretic techniques, we investigated the pink pigmentation associated with trematodiasis, infection with Podocotyloides stenometre larval trematode, in Porites compressa. Spectral emission analysis reveals that macroscopic areas of pink pigmentation fluoresce under blue light excitation (450 nm) and produce a broad emission peak at 590 nm (+/-6) with a 60-nm full width at half maximum. Electrophoretic protein separation of pigmented tissue extract confirms the red fluorescence to be a protein rather than a low-molecular-weight compound. Histological sections demonstrate green fluorescence in healthy coral tissue and red fluorescence in the trematodiasis-compromised tissue. The red fluorescent protein (FP) is limited to the epidermis, is not associated with cells or granules, and appears unstructured. These data collectively suggest that the red FP is produced and localized in tissue infected by larval trematodes and plays a role in the immune response in corals.

Fluorescence calibration method for single-particle aerosol fluorescence instruments

NASA Astrophysics Data System (ADS)

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

2017-05-01

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

An efficient core-shell fluorescent silica nanoprobe for ratiometric fluorescence detection of pH in living cells.

PubMed

Fu, Jingni; Ding, Changqin; Zhu, Anwei; Tian, Yang

2016-08-07

Intracellular pH plays a vital role in cell biology, including signal transduction, ion transport and homeostasis. Herein, a ratiometric fluorescent silica probe was developed to detect intracellular pH values. The pH sensitive dye fluorescein isothiocyanate isomer I (FITC), emitting green fluorescence, was hybridized with reference dye rhodamine B (RB), emitting red fluorescence, as a dual-emission fluorophore, in which RB was embedded in a silica core of ∼40 nm diameter. Moreover, to prevent fluorescence resonance energy transfer between FITC and RB, FITC was grafted onto the surface of core-shell silica colloidal particles with a shell thickness of 10-12 nm. The nanoprobe exhibited dual emission bands centered at 517 and 570 nm, under single wavelength excitation of 488 nm. RB encapsulated in silica was inert to pH change and only served as reference signals for providing built-in correction to avoid environmental effects. Moreover, FITC (λem = 517 nm) showed high selectivity toward H(+) against metal ions and amino acids, leading to fluorescence variation upon pH change. Consequently, variations of the two fluorescence intensities (Fgreen/Fred) resulted in a ratiometric pH fluorescent sensor. The specific nanoprobe showed good linearity with pH variation in the range of 6.0-7.8. It can be noted that the fluorescent silica probe demonstrated good water dispersibility, high stability and low cytotoxicity. Accordingly, imaging and biosensing of pH variation was successfully achieved in HeLa cells.

Fluorescent biosensor for the detection of hyaluronidase: intensity-based ratiometric sensing and fluorescence lifetime-based sensing using a long lifetime azadioxatriangulenium (ADOTA) fluorophore.

PubMed

Chib, Rahul; Mummert, Mark; Bora, Ilkay; Laursen, Bo W; Shah, Sunil; Pendry, Robert; Gryczynski, Ignacy; Borejdo, Julian; Gryczynski, Zygmunt; Fudala, Rafal

2016-05-01

In this report, we have designed a rapid and sensitive, intensity-based ratiometric sensing as well as lifetime-based sensing probe for the detection of hyaluronidase activity. Hyaluronidase expression is known to be upregulated in various pathological conditions. We have developed a fluorescent probe by heavy labeling of hyaluronic acid with a new orange/red-emitting organic azadioxatriangulenium (ADOTA) fluorophore, which exhibits a long fluorescence lifetime (∼20 ns). The ADOTA fluorophore in water has a peak fluorescence lifetime of ∼20 ns and emission spectra centered at 560 nm. The heavily ADOTA-labeled hyaluronic acid (HA-ADOTA) shows a red shift in the peak emission wavelength (605 nm), a weak fluorescence signal, and a shorter fluorescence lifetime (∼4 ns) due to efficient self-quenching and formation of aggregates. In the presence of hyaluronidase, the brightness and fluorescence lifetime of the sample increase with a blue shift in the peak emission to its original wavelength at 560 nm. The ratio of the fluorescence intensity of the HA-ADOTA probe at 560 and 605 nm can be used as the sensing method for the detection of hyaluronidase. The cleavage of the hyaluronic acid macromolecule reduces the energy migration between ADOTA molecules, as well as the degree of self-quenching and aggregation. This probe can be efficiently used for both intensity-based ratiometric sensing as well as fluorescence lifetime-based sensing of hyaluronidase. The proposed method makes it a rapid and sensitive assay, useful for analyzing levels of hyaluronidase in relevant clinical samples like urine or plasma. Graphical Abstract Scheme showing cleavage of HA-ADOTA probe by hyaluronidase and the change in the emission spectrum of HA-ADOTA probe before and after cleavage by hyaluronidase.

Variations in the endogenous fluorescence of rabbit corneas after mechanical property alterations

NASA Astrophysics Data System (ADS)

Ortega-Martinez, Antonio; Touchette, Genna; Zhu, Hong; Kochevar, Irene E.; Franco, Walfre

2017-09-01

Keratoconus is an eye disease in which the cornea progressively deforms due to loss of cornea mechanical rigidity, and thus causes deterioration of visual acuity. Techniques to characterize the mechanical characteristics of the cornea are important to better monitor changes and response to treatments. To investigate the feasibility of using the endogenous fluorescence of cornea for monitoring alterations of its mechanical rigidity, linear tensiometry was used to quantitate stiffness and Young's modulus (YM) after treatments that increase cornea stiffness (collagen photocross-linking) or decrease stiffness (enzymatic digestion). The endogenous ultraviolet fluorescence of cornea was also measured before and after these treatments. The fluorescence excitation/emission spectral ranges were 280 to 430/390 to 520 nm, respectively. A correlation analysis was carried out to identify fluorescence excitation/emission pairs whose intensity changes correlated with the stiffness. A positive correlation was found between variations in fluorescence intensity of the 415-/485-nm excitation/emission pair and YM of photocross-linked corneas. After treatment of corneas with pepsin, the YM decreased as the fluorescence intensity at 290-/390-nm wavelengths decreased. For weakening of corneas with collagenase, only qualitative changes in the fluorescence spectrum were observed. Changes in the concentration of native or newly created fluorescent molecular species contain information that may be directly or indirectly related to the mechanical structure of the cornea.

A New Thiosemicarbazone-Based Fluorescence "Turn-on" Sensor for Zn(2+) Recognition with a Large Stokes Shift and its Application in Live Cell Imaging.

PubMed

Tang, Lijun; Huang, Zhenlong; Zheng, Zhuxuan; Zhong, Keli; Bian, Yanjiang

2016-09-01

Selective fluorescence turn on Zn(2+) sensor with long-wavelength emission and a large Stokes shift is highly desirable in Zn(2+) sensing area. We reported herein the synthesis and Zn(2+) recognition properties of a new thiosemicarbazone-based fluorescent sensor L. L displays high selectivity and sensitivity toward Zn(2+) over other metal ions in DMSO-H2O (1:1, v/v, HEPES 10 mM, pH = 7.4) solution with a long-wavelength emission at 572 nm and a large Stokes shift of 222 nm. Confocal fluorescence microscopy experiments demonstrate that L is cell-permeable and capable of monitoring intracellular Zn(2+). Graphical Abstract We report a new thiosemicarbazone-based fluorescent sensor (L) for selective recognition of Zn(2+) with a long wavelength emission and a large Stokes shift.

DNA nanostructure-based fluorescence thermometer with silver nanoclusters

NASA Astrophysics Data System (ADS)

Bu, Congcong; Mu, Lixuan; Cao, Xingxing; Chen, Min; She, Guangwei; Shi, Wensheng

2018-07-01

DNA nanostructure-based fluorescence thermometers were fabricated by linking fluorescent silver nanoclusters (AgNCs) and guanine-rich（G-rich）DNA chains via a thermally sensitive DNA stem-loop at terminals 5‧ and 3‧. Variations of temperature alter the distance between the AgNCs and G-rich DNA chain, affecting the interaction between them. As a result, the intensity of fluorescence emission from the AgNCs at 636 nm can be sensitively modulated. It was found that the intensity of such red emission is more temperature sensitive than the equivalent green emission at 543 nm; sensitivity of ‑3.6%/°C was achieved. Through variation of the melting temperature of the DNA stem-loop, the response temperature range of the thermometers could be readily adjusted. Novel DNA nanostructure-based fluorescence thermometers as described in this work are anticipated to be able to measure the temperature of biological systems at small scales—even a single cell.

DNA nanostructure-based fluorescence thermometer with silver nanoclusters.

PubMed

Bu, Congcong; Mu, Lixuan; Cao, XIngxing; Chen, Min; She, Guangwei; Shi, Wensheng

2018-04-27

Linking the fluorescent silver nanoclusters (AgNCs) and guanine-rich（G-rich）DNA chains by the thermal sensitive DNA stem-loop at teminal 5' and 3', DNA nanostructure-based fluorescence thermometers were fabricated. The variations of the temperature alter the distance between AgNCs and G-rich DNA chain, which could affect the interaction between them. As a result, the intensity of fluorescence emission from AgNCs at 636 nm can be sensitively modulated. It was found that such red emission is more sensitive to the temperature comparing with its intrinsic green emission at 543 nm, and sensitivity of -3.6%/℃ was achieved. Varying the melting temperature of the DNA stem-loop could readily adjust the response temperature range of thermometers. Novel DNA nanostructure-based fluorescence thermometers in this work could be anticipated to measure the temperature of biological system, even a single cell. © 2018 IOP Publishing Ltd.

Fluorescence of silicon nanoparticles prepared by nanosecond pulsed laser

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

Liu, Chunyang, E-mail: chunyangliu@126.com; Sui, Xin; Yang, Fang

2014-03-15

A pulsed laser fabrication method is used to prepare fluorescent microstructures on silicon substrates in this paper. A 355 nm nanosecond pulsed laser micromachining system was designed, and the performance was verified and optimized. Fluorescence microscopy was used to analyze the photoluminescence of the microstructures which were formed using the pulsed laser processing technique. Photoluminescence spectra of the microstructure reveal a peak emission around 500 nm, from 370 nm laser irradiation. The light intensity also shows an exponential decay with irradiation time, which is similar to attenuation processes seen in porous silicon. The surface morphology and chemical composition of themore » microstructure in the fabricated region was also analyzed with multifunction scanning electron microscopy. Spherical particles are produced with diameters around 100 nm. The structure is compared with porous silicon. It is likely that these nanoparticles act as luminescence recombination centers on the silicon surface. The small diameter of the particles modifies the band gap of silicon by quantum confinement effects. Electron-hole pairs recombine and the fluorescence emission shifts into the visible range. The chemical elements of the processed region are also changed during the interaction between laser and silicon. Oxidation and carbonization play an important role in the enhancement of fluorescence emission.« less

Intraoperative delineation of primary brain tumors using time-resolved fluorescence spectroscopy

PubMed Central

Butte, Pramod V.; Fang, Qiyin; Jo, Javier A.; Yong, William H.; Pikul, Brian K.; Black, Keith L.; Marcu, Laura

2010-01-01

The goal of this study is to determine the potential of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) as an adjunctive tool for delineation of brain tumor from surrounding normal tissue in order to assist the neurosurgeon in near-complete tumor excision. A time-domain TR-LIFS prototype apparatus (gated photomultiplier detection, fast digitizer) was used for recording tissue autofluorescence in normal cortex (NC), normal white matter (NWM), and various grades of gliomas intraoperatively. Tissue fluorescence was induced with a pulsed nitrogen laser (337nm, 700ps), and the intensity decay profiles were recorded in the 360-to550-nm spectral range (10-nm interval). Histopathological analysis (hematoxylin & eosin) of the biopsy samples taken from the site of TR-LIFS measurements was used for validation of spectroscopic results. Preliminary results on 17 patients demonstrate that normal cortex (N=16) and normal white matter (N=3) show two peaks of fluorescence emission at 390nm(lifetime=1.8±0.3ns) and 460nm(lifetime=0.8±0.1ns). The 390-nm emission peak is absent in low-grade glioma (N=5; lifetime=1.1ns) and reduced in high-grade glioma (N=9; lifetime=1.7±0.4ns). The emission characteristics at 460nm in all tissues correlated with the nicotinamide adenine dinucleotide fluorescence (peak: 440to460nm; lifetime: 0.8to1.0ns). These findings demonstrate the potential of using TR-LIFS as a tool for enhanced delineation of brain tumors during surgery. In addition, this study evaluates similarities and differences between TR-LIFS signatures of brain tumors obtained in vivo and those previously reported in ex vivo brain tumor specimens. PMID:20459282

Intraoperative delineation of primary brain tumors using time-resolved fluorescence spectroscopy.

PubMed

Butte, Pramod V; Fang, Qiyin; Jo, Javier A; Yong, William H; Pikul, Brian K; Black, Keith L; Marcu, Laura

2010-01-01

The goal of this study is to determine the potential of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) as an adjunctive tool for delineation of brain tumor from surrounding normal tissue in order to assist the neurosurgeon in near-complete tumor excision. A time-domain TR-LIFS prototype apparatus (gated photomultiplier detection, fast digitizer) was used for recording tissue autofluorescence in normal cortex (NC), normal white matter (NWM), and various grades of gliomas intraoperatively. Tissue fluorescence was induced with a pulsed nitrogen laser (337 nm, 700 ps), and the intensity decay profiles were recorded in the 360- to 550-nm spectral range (10-nm interval). Histopathological analysis (hematoxylin & eosin) of the biopsy samples taken from the site of TR-LIFS measurements was used for validation of spectroscopic results. Preliminary results on 17 patients demonstrate that normal cortex (N=16) and normal white matter (N=3) show two peaks of fluorescence emission at 390 nm (lifetime=1.8+/-0.3 ns) and 460 nm (lifetime=0.8+/-0.1 ns). The 390-nm emission peak is absent in low-grade glioma (N=5; lifetime=1.1 ns) and reduced in high-grade glioma (N=9; lifetime=1.7+/-0.4 ns). The emission characteristics at 460 nm in all tissues correlated with the nicotinamide adenine dinucleotide fluorescence (peak: 440 to 460 nm; lifetime: 0.8 to 1.0 ns). These findings demonstrate the potential of using TR-LIFS as a tool for enhanced delineation of brain tumors during surgery. In addition, this study evaluates similarities and differences between TR-LIFS signatures of brain tumors obtained in vivo and those previously reported in ex vivo brain tumor specimens.

Intraoperative delineation of primary brain tumors using time-resolved fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Butte, Pramod V.; Fang, Qiyin; Jo, Javier A.; Yong, William H.; Pikul, Brian K.; Black, Keith L.; Marcu, Laura

2010-03-01

The goal of this study is to determine the potential of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) as an adjunctive tool for delineation of brain tumor from surrounding normal tissue in order to assist the neurosurgeon in near-complete tumor excision. A time-domain TR-LIFS prototype apparatus (gated photomultiplier detection, fast digitizer) was used for recording tissue autofluorescence in normal cortex (NC), normal white matter (NWM), and various grades of gliomas intraoperatively. Tissue fluorescence was induced with a pulsed nitrogen laser (337 nm, 700 ps), and the intensity decay profiles were recorded in the 360- to 550-nm spectral range (10-nm interval). Histopathological analysis (hematoxylin & eosin) of the biopsy samples taken from the site of TR-LIFS measurements was used for validation of spectroscopic results. Preliminary results on 17 patients demonstrate that normal cortex (N=16) and normal white matter (N=3) show two peaks of fluorescence emission at 390 nm (lifetime=1.8+/-0.3 ns) and 460 nm (lifetime=0.8+/-0.1 ns). The 390-nm emission peak is absent in low-grade glioma (N=5; lifetime=1.1 ns) and reduced in high-grade glioma (N=9; lifetime=1.7+/-0.4 ns). The emission characteristics at 460 nm in all tissues correlated with the nicotinamide adenine dinucleotide fluorescence (peak: 440 to 460 nm lifetime: 0.8 to 1.0 ns). These findings demonstrate the potential of using TR-LIFS as a tool for enhanced delineation of brain tumors during surgery. In addition, this study evaluates similarities and differences between TR-LIFS signatures of brain tumors obtained in vivo and those previously reported in ex vivo brain tumor specimens.

In vitro energy transfer in Renilla bioluminescence

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

Ward, W.W.; Cormier, M.J.

1976-09-23

A quantitative study of in vitro energy transfer in a natural biological system is reported. The in vitro bioluminescent oxidation of Renilla (sea pansy) luciferin by luciferase produces a broad, structureless emission, peaking in the blue at 490 nm. In contrast, the live animal produces a structured emission peaking in the green at 509 nm. This difference in emission characteristics is due to the presence, in Renilla, of a green fluorescent protein (GFP). Addition of GFP in vitro sensitizes the oxyluciferin product excited state, resulting in the narrow, structured green emission characteristic of GFP fluorescence (lambda/sub max/ 509 nm). Undermore » conditions of efficient in vitro energy transfer (2.7 x 10/sup -6/ M GFP) the radiative quantum yield (with respect to luciferin) increases 5.7-fold from 5.3% (blue pathway) to 30% (green pathway). The fluorescence quantum yield of the Renilla GFP has been measured as 30%; thus, within the precision of our measurements (15% coefficient of variation) the in vitro energy transfer efficiency is a surprising 100%.« less

Red Fluorescent Carbon Nanoparticle-Based Cell Imaging Probe.

PubMed

Ali, Haydar; Bhunia, Susanta Kumar; Dalal, Chumki; Jana, Nikhil R

2016-04-13

Fluorescent carbon nanoparticle-based probes with tunable visible emission are biocompatible, environment friendly and most suitable for various biomedical applications. However, synthesis of red fluorescent carbon nanoparticles and their transformation into functional nanoparticles are very challenging. Here we report red fluorescent carbon nanoparticle-based nanobioconjugates of <25 nm hydrodynamic size and their application as fluorescent cell labels. Hydrophobic carbon nanoparticles are synthesized via high temperature colloid-chemical approach and transformed into water-soluble functional nanoparticles via coating with amphiphilic polymer followed by covalent linking with desired biomolecules. Following this approach, carbon nanoparticles are functionalized with polyethylene glycol, primary amine, glucose, arginine, histidine, biotin and folic acid. These functional nanoparticles can be excited with blue/green light (i.e., 400-550 nm) to capture their emission spanning from 550 to 750 nm. Arginine and folic acid functionalized nanoparticles have been demonstrated as fluorescent cell labels where blue and green excitation has been used for imaging of labeled cells. The presented method can be extended for the development of carbon nanoparticle-based other bioimaging probes.

Insight into the heterogeneous adsorption of humic acid fluorescent components on multi-walled carbon nanotubes by excitation-emission matrix and parallel factor analysis.

PubMed

Yang, Chenghu; Liu, Yangzhi; Cen, Qiulin; Zhu, Yaxian; Zhang, Yong

2018-02-01

The heterogeneous adsorption behavior of commercial humic acid (HA) on pristine and functionalized multi-walled carbon nanotubes (MWCNTs) was investigated by fluorescence excitation-emission matrix and parallel factor (EEM- PARAFAC) analysis. The kinetics, isotherms, thermodynamics and mechanisms of adsorption of HA fluorescent components onto MWCNTs were the focus of the present study. Three humic-like fluorescent components were distinguished, including one carboxylic-like fluorophore C1 (λ ex /λ em = (250, 310) nm/428nm), and two phenolic-like fluorophores, C2 (λ ex /λ em = (300, 460) nm/552nm) and C3 (λ ex /λ em = (270, 375) nm/520nm). The Lagergren pseudo-second-order model can be used to describe the adsorption kinetics of the HA fluorescent components. In addition, both the Freundlich and Langmuir models can be suitably employed to describe the adsorption of the HA fluorescent components onto MWCNTs with significantly high correlation coefficients (R 2 > 0.94, P< 0.05). The dissimilarity in the adsorption affinity (K d ) and nonlinear adsorption degree from the HA fluorescent components to MWCNTs was clearly observed. The adsorption mechanism suggested that the π-π electron donor-acceptor (EDA) interaction played an important role in the interaction between HA fluorescent components and the three MWCNTs. Furthermore, the values of the thermodynamic parameters, including the Gibbs free energy change (ΔG°), enthalpy change (ΔH°) and entropy change (ΔS°), showed that the adsorption of the HA fluorescent components on MWCNTs was spontaneous and exothermic. Copyright © 2017 Elsevier Inc. All rights reserved.

Multispectral laser-induced fluorescence imaging system for large biological samples

NASA Astrophysics Data System (ADS)

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

2003-07-01

A laser-induced fluorescence imaging system developed to capture multispectral fluorescence emission images simultaneously from a relatively large target object is described. With an expanded, 355-nm Nd:YAG laser as the excitation source, the system captures fluorescence emission images in the blue, green, red, and far-red regions of the spectrum centered at 450, 550, 678, and 730 nm, respectively, from a 30-cm-diameter target area in ambient light. Images of apples and of pork meat artificially contaminated with diluted animal feces have demonstrated the versatility of fluorescence imaging techniques for potential applications in food safety inspection. Regions of contamination, including sites that were not readily visible to the human eye, could easily be identified from the images.

Preparation and flow cytometry of uniform silica-fluorescent dye microspheres.

PubMed

Bele, Marjan; Siiman, Olavi; Matijević, Egon

2002-10-15

Uniform fluorescent silica-dye microspheres have been prepared by coating preformed monodispersed silica particles with silica layers containing rhodamine 6G or acridine orange. The resulting dispersions exhibit intense fluorescent emission between 500 and 600 nm, over a broad excitation wavelength range of 460 to 550 nm, even with exceedingly small amounts of dyes incorporated into the silica particles (10-30 ppm, expressed as weight of dye relative to weight of dry particles). The fluorescent particles can be prepared in micrometer diameters suitable for analyses using flow cytometry with 488-nm laser excitation.

Pathological changes in Alzheimer"s brain evaluated with fluorescence emission analysis (FEA)

NASA Astrophysics Data System (ADS)

Christov, Alexander; Ottman, Todd; Grammas, Paula

2004-07-01

Development of AD is associated with cerebrovascular deposition of amyloid beta (Aβ) as well as a progressive increase in vasular collagen content. Both AΒ and collagen are naturally fluorescent compounds when exposed to UV light. We analyzed autofluorescence emitted from brain tissue samples and isolated brain resistance vessels harvested postmortem from patients with Alzheimer's disease (AD) and age-matched controls. Fluorescence emission, excited at 355 nm with an Nd:YAG laser, was measured using a fiber-optic based fluorescence spectroscopic system for tissue analysis. Significantly higher values of fluorescence emission intensity (P<0.001) in the spectral region from 465 to 490 nm were detected in brain resistance vessel samples from AD patients compared to the normal individuals. Results from western blot analysis showed elevated levels of type I and type III collagen, and reduced levels of type IV collagen in resistance vessels from AD patients, compared to control samples. In addition, using direct scanning of the cortical suface for fluoresxcence emission by the laser-induced fluorescence spectroscopy system we detected a significantly (P<0.05) higher level of apoptosis in AD brain tissue compared to age-matched controls. Fluorescence emission analysis (FEA) appears to be a sensitive technique for detecting structural changes in AD brain tissue.

Fluorescence lifetime spectroscopy for guided therapy of brain tumors.

PubMed

Butte, Pramod V; Mamelak, Adam N; Nuno, Miriam; Bannykh, Serguei I; Black, Keith L; Marcu, Laura

2011-01-01

This study evaluates the potential of time-resolved laser induced fluorescence spectroscopy (TR-LIFS) as intra-operative tool for the delineation of brain tumor from normal brain. Forty two patients undergoing glioma (WHO grade I-IV) surgery were enrolled in this study. A TR-LIFS prototype apparatus (gated detection, fast digitizer) was used to induce in-vivo fluorescence using a pulsed N2 laser (337 nm excitation, 0.7 ns pulse width) and to record the time-resolved spectrum (360-550 nm range, 10 nm interval). The sites of TR-LIFS measurement were validated by conventional histopathology (H&E staining). Parameters derived from the TR-LIFS data including intensity values and time-resolved intensity decay features (average fluorescence lifetime and Laguerre coefficients values) were used for tissue characterization and classification. 71 areas of tumor and normal brain were analyzed. Several parameters allowed for the differentiation of distinct tissue types. For example, normal cortex (N=35) and normal white matter (N=12) exhibit a longer-lasting fluorescence emission at 390 nm (τ390=2.12±0.10 ns) when compared with 460 nm (τ460=1.16±0.08 ns). High grade glioma (grades III and IV) samples (N=17) demonstrate emission peaks at 460 nm, with large variation at 390 nm while low grade glioma (I and II) samples (N=7) demonstrated a peak fluorescence emission at 460 nm. A linear discriminant algorithm allowed for the classification of low-grade gliomas with 100% sensitivity and 98% specificity. High-grade glioma demonstrated a high degree of heterogeneity thus reducing the discrimination accuracy of these tumors to 47% sensitivity and 94% specificity. Current findings demonstrate that TR-LIFS holds the potential to diagnose brain tumors intra-operatively and to provide a valuable tool for aiding the neurosurgeon-neuropathologist team in to rapidly distinguish between tumor and normal brain during surgery. Copyright © 2010 Elsevier Inc. All rights reserved.

An analysis of dissolved organic matter from freshwater Karelian Lakes using reversed-phase high-performance liquid chromatography with online absorbance and fluorescence analysis

NASA Astrophysics Data System (ADS)

Khundzhua, D. A.; Patsaeva, S. V.; Trubetskoj, O. A.; Trubetskaya, O. E.

2017-01-01

The spectral and optical properties of the fractionated components of dissolved organic matter (DOM) of three freshwater lakes in Karelia were studied using reversed-phase high-performance liquid chromatography (RP-HPLC) with online detection of fluorescence and absorption spectra. It is shown that the DOM fractions are qualitatively similar, but differ quantitatively in the ratio of components and consist of at least three types of fluorophores: (1) hydrophilic "humic-like" fluorophore(s) with the emission maximum in the region of 420 nm and an absorption band at 260-270 nm; (2) hydrophobic "humic-like" fluorophore(s) with the emission maximum at approximately 450 nm that has no characteristic absorption maxima in the region from 220 to 400 nm; and (3) a "protein-like" fluorophore with the emission maximum in the region of 340-350 nm, which is typical of proteins and peptides containing tryptophan.

A simple one-step method to prepare fluorescent carbon dots and their potential application in non-invasive glioma imaging

NASA Astrophysics Data System (ADS)

Ruan, Shaobo; Qian, Jun; Shen, Shun; Zhu, Jianhua; Jiang, Xinguo; He, Qin; Gao, Huile

2014-08-01

Fluorescent carbon dots (CD) possess impressive potential in bioimaging because of their low photobleaching, absence of optical blinking and good biocompatibility. However, their relatively short excitation/emission wavelengths restrict their application in in vivo imaging. In the present study, a kind of CD was prepared by a simple heat treatment method using glycine as the only precursor. The diameter of CD was lower than 5 nm, and the highest emission wavelength was 500 nm. However, at 600 nm, there was still a relatively strong fluorescent emission, suggesting CD could be used for in vivo imaging. Additionally, several experiments demonstrated that CD possessed good serum stability and low cytotoxicity. In vitro, CD could be taken up into C6 glioma cells in a time- and concentration-dependent manner, with both endosomes and mitochondria involved. In vivo, CD could be used for non-invasive glioma imaging because of its high accumulation in the glioma site of the brain, which was demonstrated by both in vivo imaging and ex vivo tissue imaging. Furthermore, the fluorescent distribution in tissue slices also showed CD distributed in glioma with high intensity, while with a low intensity in normal brain tissue. In conclusion, CD were prepared using a simple method with relatively long excitation and emission wavelengths and could be used for non-invasive glioma imaging.Fluorescent carbon dots (CD) possess impressive potential in bioimaging because of their low photobleaching, absence of optical blinking and good biocompatibility. However, their relatively short excitation/emission wavelengths restrict their application in in vivo imaging. In the present study, a kind of CD was prepared by a simple heat treatment method using glycine as the only precursor. The diameter of CD was lower than 5 nm, and the highest emission wavelength was 500 nm. However, at 600 nm, there was still a relatively strong fluorescent emission, suggesting CD could be used for in vivo imaging. Additionally, several experiments demonstrated that CD possessed good serum stability and low cytotoxicity. In vitro, CD could be taken up into C6 glioma cells in a time- and concentration-dependent manner, with both endosomes and mitochondria involved. In vivo, CD could be used for non-invasive glioma imaging because of its high accumulation in the glioma site of the brain, which was demonstrated by both in vivo imaging and ex vivo tissue imaging. Furthermore, the fluorescent distribution in tissue slices also showed CD distributed in glioma with high intensity, while with a low intensity in normal brain tissue. In conclusion, CD were prepared using a simple method with relatively long excitation and emission wavelengths and could be used for non-invasive glioma imaging. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr02657h

Dy3+ doped tellurite glasses containing silver nanoparticles for lighting devices

NASA Astrophysics Data System (ADS)

Hua, Chenxiao; Shen, Lifan; Pun, Edwin Yue Bun; Li, Desheng; Lin, Hai

2018-04-01

Efficient warm yellowish-white fluorescence emissions of Dy3+ were observed in heavy metal germanium tellurite (HGT) glasses under the excitation of 454 nm. Further, the luminescence intensity of Dy3+ is increased by ∼29% accompanying the introduction of Ag NPs with diameter ∼7 nm when compared with that of the silver-free case, which is caused by the existence of localized surface plasmon resonance (LSPR). The larger net emission power, the more net emission photon number and the higher quantum yield in Dy2O3 doped HGT glasses containing Ag NPs (HGT-Ag) confirm the availability of utilizing laser. Presupposed fluorescence color trace reveals that white luminescence can be achieved when the intensity ratio between residual laser and Dy3+ emission reaches the appropriate range. The productive transition emissions and the tunable white fluorescence illustrate tellurite glasses embodying noble-metal NPs are a potential candidate for high-quality lighting devices.

Authentication of the botanical origin of honey by front-face fluorescence spectroscopy. A preliminary study.

PubMed

Ruoff, Kaspar; Karoui, Romdhane; Dufour, Eric; Luginbühl, Werner; Bosset, Jacques-Olivier; Bogdanov, Stefan; Amado, Renato

2005-03-09

The potential of front-face fluorescence spectroscopy for the authentication of unifloral and polyfloral honey types (n = 57 samples) previously classified using traditional methods such as chemical, pollen, and sensory analysis was evaluated. Emission spectra were recorded between 280 and 480 nm (excit: 250 nm), 305 and 500 nm (excit: 290 nm), and 380 and 600 nm (excit: 373 nm) directly on honey samples. In addition, excitation spectra (290-440 nm) were recorded with the emission measured at 450 nm. A total of four different spectral data sets were considered for data analysis. After normalization of the spectra, chemometric evaluation of the spectral data was carried out using principal component analysis (PCA) and linear discriminant analysis (LDA). The rate of correct classification ranged from 36% to 100% by using single spectral data sets (250, 290, 373, 450 nm) and from 73% to 100% by combining these four data sets. For alpine polyfloral honey and the unifloral varieties investigated (acacia, alpine rose, honeydew, chestnut, and rape), correct classification ranged from 96% to 100%. This preliminary study indicates that front-face fluorescence spectroscopy is a promising technique for the authentication of the botanical origin of honey. It is nondestructive, rapid, easy to use, and inexpensive. The use of additional excitation wavelengths between 320 and 440 nm could increase the correct classification of the less characteristic fluorescent varieties.

Fluorescence spectroscopy for rapid detection and classification of bacterial pathogens.

PubMed

Sohn, Miryeong; Himmelsbach, David S; Barton, Franklin E; Fedorka-Cray, Paula J

2009-11-01

This study deals with the rapid detection and differentiation of Escherichia coli, Salmonella, and Campylobacter, which are the most commonly identified commensal and pathogenic bacteria in foods, using fluorescence spectroscopy and multivariate analysis. Each bacterial sample cultured under controlled conditions was diluted in physiologic saline for analysis. Fluorescence spectra were collected over a range of 200-700 nm with 0.5 nm intervals on the PerkinElmer Fluorescence Spectrometer. The synchronous scan technique was employed to find the optimum excitation (lambda(ex)) and emission (lambda(em)) wavelengths for individual bacteria with the wavelength interval (Deltalambda) being varied from 10 to 200 nm. The synchronous spectra and two-dimensional plots showed two maximum lambda(ex) values at 225 nm and 280 nm and one maximum lambda(em) at 335-345 nm (lambda(em) = lambda(ex) + Deltalambda), which correspond to the lambda(ex) = 225 nm, Deltalambda = 110-120 nm, and lambda(ex) = 280 nm, Deltalambda = 60-65 nm. For all three bacterial genera, the same synchronous scan results were obtained. The emission spectra from the three bacteria groups were very similar, creating difficulty in classification. However, the application of principal component analysis (PCA) to the fluorescence spectra resulted in successful classification of the bacteria by their genus as well as determining their concentration. The detection limit was approximately 10(3)-10(4) cells/mL for each bacterial sample. These results demonstrated that fluorescence spectroscopy, when coupled with PCA processing, has the potential to detect and to classify bacterial pathogens in liquids. The methodology is rapid (>10 min), inexpensive, and requires minimal sample preparation compared to standard analytical methods for bacterial detection.

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

NASA Astrophysics Data System (ADS)

Makoui, Anali

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

Investigation of formalin influence over hard and soft biological tissues fluorescent spectra in vitro

NASA Astrophysics Data System (ADS)

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

2007-05-01

In order to investigate the formalin influence over fluorescence properties of hard and soft biological tissues during conservation, emission spectra have been registered. Nitrogen laser at 337 nm and light-emitting diode with maximum at 405 nm have been used as excitation sources. For investigation of formalin influence over hard tissues, an experiment was made on teeth samples. Sound teeth were demineralized with a phosphoric acid for 10 seconds to obtain enamel structure near to the tooth lesion, and were fixed in formalin. Before and after teeth treatment spectra from the areas of interest were detected. There were not observed changes in the shape of the teeth spectra, related to the introduction of formalin fluorescence. Samples from mucosa of esophagus and stomach, where initially an ALA/Protoporphyrin IX diagnosis was applied, were used as soft tissue specimens. After fluorescent diagnosis in vivo biopsy samples were obtained from normal and cancerous areas and were conserved in formalin. Initially, spectrum observed has one autofluorescence maximum from the mucous tissue at 500-600 nm and secondary maxima from the protoporphyrin fluorescence at 635 nm and 720 nm, as well as pronounced minima at 540 and 575 nm related to hemoglobin absorption. After formalin conservation hemoglobin absorption was strongly reduced that increases mucous emission signal in green-yellow spectral region. Simultaneously the maxima at 635 nm and 720 nm were reduced. As conclusion we could say that formalin has negligible influence over fluorescence spectra of conserved hard tissues and has more pronounced influence over fluorescence spectra obtained in the case of soft tissue conservation, which has to be taking into account in measurements in vitro.

Violet and blue light-induced green fluorescence emissions from dental caries.

PubMed

Shakibaie, F; Walsh, L J

2016-12-01

The objective of this laboratory study was to compare violet and visible blue LED light-elicited green fluorescence emissions from enamel and dentine in healthy or carious states. Microscopic digital photography was undertaken using violet and blue LED illumination (405 nm and 455 nm wavelengths) of tooth surfaces, which were photographed through a custom-made stack of green compensating filters which removed the excitation light and allowed green fluorescence emissions to pass. Green channel pixel data were analysed. Dry sound enamel and sound root surfaces showed strong green fluorescence when excited by violet or blue lights. Regions of cavitated dental caries gave lower green fluorescence, and this was similar whether the dentine in the lesions was the same colour as normal dentine or was darkly coloured. The presence of saliva on the surface did not significantly change the green fluorescence, while the presence of blood diluted in saliva depressed green fluorescence. Using violet or blue illumination in combination with green compensating filters could potentially aid in the assessment of areas of mineral loss. © 2016 Australian Dental Association.

A direct and simultaneous detection of zinc protoporphyrin IX, free protoporphyrin IX, and fluorescent heme degradation product in red blood cell hemolysates.

PubMed

Chen, Qiuying; Hirsch, Rhoda Elison

2006-03-01

Fluorescence emission of free protoporphyrin IX (PPIX, em. approximately 626 nm), zinc protoporphyrin IX (ZPP, em. approximately 594 nm) and fluorescent heme degradation product (FHDP, em. approximately 466 nm) are identified and simultaneously detected in mouse and human red cell hemolysates, when excited at 365 nm. A novel method is established for comparing relative FHDP, PPIX and ZPP levels in hemolysates without performing red cell porphyrin extractions. The ZPP fluorescence directly measured in hemolysates (F(365/594)) correlates with the ZPP fluorescence obtained from acetone/water extraction (R(2) = 0.9515, P < 0.0001). The relative total porphyrin (ZPP and PPIX) fluorescence obtained from direct hemolysate fluorescence measurements also correlates with red blood cell total porphyrins determined by ethyl acetate extraction (Piomelli extraction, R(2) = 0.88, P < 0.0001). These fluorescent species serves as biomarkers for alterations in Hb synthesis and Hb stability.

Spectroscopy of high index contrast Yb:Ta2O5 waveguides for lasing applications

NASA Astrophysics Data System (ADS)

Aghajani, A.; Murugan, G. S.; Sessions, N. P.; Apostolopoulos, V.; Wilkinson, J. S.

2015-06-01

Ytterbium-doped waveguides are required for compact integrated lasers and Yb- doped Ta2O5 is a promising candidate material. The design, fabrication and spectroscopic characterisation of Yb:Ta2O5 rib waveguides are described. The peak absorption cross-section was measured to be 2.75×10-20 cm2 at 975 nm. The emission spectrum was found to have a fluorescence emission peak at a wavelength of 976 nm with a peak cross-section of 2.9×10-20 cm2 and a second broad fluorescence band spanning from 990 nm to 1090 nm. The excited- state life time was measured to be 260 μs.

A ratiometric fluorescent probe for hydrophobic proteins in aqueous solution based on aggregation-induced emission.

PubMed

Peng, Lu; Wei, Ruirui; Li, Kai; Zhou, Zhaojuan; Song, Panshu; Tong, Aijun

2013-04-07

A novel fluorescent probe 1 is reported here with ratiometric response to hydrophobic proteins (casein) or proteins with hydrophobic pockets (BSA, HSA) through hydrophobic interaction. Probe 1 underwent deprotonation in aqueous solution at pH 7.4 and emitted blue fluorescence at 436 nm. Upon the addition of BSA, HSA or casein, the aggregation-induced emission fluorescence of 1 at 518 nm was turned on. The fluorescence intensity ratio, I518/I436 was linearly related to the concentrations of these proteins. The detection limits for BSA, HSA and casein based on IUPAC (CDL = 3Sb m(-1)) were 16.2 μg mL(-1), 10.5 μg mL(-1) and 5.7 μg mL(-1), respectively.

Droplet-based immunoassay on a 'sticky' nanofibrous surface for multiplexed and dual detection of bacteria using smartphones.

PubMed

Nicolini, Ariana M; Fronczek, Christopher F; Yoon, Jeong-Yeol

2015-05-15

We have developed a rapid, sensitive, and specific droplet-based immunoassay for the detection of Escherichia coli and Salmonella within a single-pipetted sample. Polycaprolactone (PCL) electrospun fibers on indium-tin-oxide (ITO) glass provide a sufficient surface to render a non-slip droplet condition, and while the PCL fibers lend a local hydrophilicity (contact angle θ=74°) for sufficient sub-micron particle adhesion, air pockets within the fibers lend an apparent hydrophobicity. Overall, the contact angle of water on this electrospun surface is 119°, and the air pockets cause the droplet to be completely immobile and resistant to movement, protecting it from external vibration. By using both anti-E. coli conjugated, 510 nm diameter green fluorescent particles (480 nm excitation and 520 nm emission) and anti-Salmonella conjugated, 400 nm diameter red fluorescent particles (640 nm excitation and 690 nm emission), we can detect multiple targets in a single droplet. Using appropriate light sources guided by fiber optics, we determined a detection limit of 10(2) CFU mL(-1). Immunoagglutination can be observed under a fluorescence microscope. Fluorescence detection (at the emission wavelength) of immunoagglutination was maximum at 90° from the incident light, while light scattering (at the excitation wavelength) was still present and behaved similarly, indicating the ability of double detection, greatly improving credibility and reproducibility of the assay. A power function (light intensity) simulation of elastic Mie scatter confirmed that both fluorescence and light scattering were present. Due to the size of the fluorescent particles relative to their incident excitation wavelengths, Mie scatter conditions were observed, and fluorescence signals show a similar trend to light scattering signals. Smartphone detection was included for true portable detection, in which the high contact angle pinning of the droplet makes this format re-usable and re-configurable. Copyright © 2014 Elsevier B.V. All rights reserved.

Investigating temperature effects on extra virgin olive oil using fluorescence spectroscopy

NASA Astrophysics Data System (ADS)

Saleem, M.; Ahmad, Naveed; Ali, H.; Bilal, M.; Khan, Saranjam; Ullah, Rahat; Ahmed, M.; Mahmood, S.

2017-12-01

The potential of fluorescence spectroscopy has been utilized to study the heating effects on extra virgin olive oil (EVOO). Through a series of experiments, a temperature range from 140 °C  -  150 °C has been found where cooking with EVOO is possible without destroying its natural ingredients. Fluorescence emission spectra from all heated and non-heated EVOO samples were recorded using an excitation source at 350 nm, where emission bands in non-heated EVOO at 380, 440, 455, and 525 nm are labelled for vitamin E and a band at 673 nm is assigned for chlorophyll a. The emission band at 525 nm is also responsible for beta carotenoids (vitamin A). As a result of heating, prominent intensity variations have been observed in all spectral bands, but it is particularly affected at 525 nm, indicating the deterioration of vitamin E and beta carotenoids. However, if the temperature of oil can be maintained in the above defined range, then frying food with EVOO is possible by preserving its natural ingredients. The spectral variations resulting from the heating effects have been further highlighted by using principal component analysis for classification purposes.

Label-Free Carbon-Dots-Based Ratiometric Fluorescence pH Nanoprobes for Intracellular pH Sensing.

PubMed

Shangguan, Jingfang; He, Dinggeng; He, Xiaoxiao; Wang, Kemin; Xu, Fengzhou; Liu, Jinquan; Tang, Jinlu; Yang, Xue; Huang, Jin

2016-08-02

Measuring pH in living cells is of great importance for better understanding cellular functions as well as providing pivotal assistance for early diagnosis of diseases. In this work, we report the first use of a novel kind of label-free carbon dots for intracellular ratiometric fluorescence pH sensing. By simple one-pot hydrothermal treatment of citric acid and basic fuchsin, the carbon dots showing dual emission bands at 475 and 545 nm under single-wavelength excitation were synthesized. It is demonstrated that the fluorescence intensities of the as-synthesized carbon dots at the two emissions are pH-sensitive simultaneously. The intensity ratio (I475 nm/I545 nm) is linear against pH values from 5.2 to 8.8 in buffer solution, affording the capability as ratiometric probes for intracellular pH sensing. It also displays that the carbon dots show excellent reversibility and photostability in pH measurements. With this nanoprobe, quantitative fluorescence imaging using the ratio of two emissions (I475 nm/I545 nm) for the detection of intracellular pH were successfully applied in HeLa cells. In contrast to most of the reported nanomaterials-based ratiometric pH sensors which rely on the attachment of additional dyes, these carbon-dots-based ratiometric probes are low in toxicity, easy to synthesize, and free from labels.

Double-excitation fluorescence spectral imaging: eliminating tissue auto-fluorescence from in vivo PPIX measurements

NASA Astrophysics Data System (ADS)

Torosean, Sason; Flynn, Brendan; Samkoe, Kimberley S.; Davis, Scott C.; Gunn, Jason; Axelsson, Johan; Pogue, Brian W.

2012-02-01

An ultrasound coupled handheld-probe-based optical fluorescence molecular tomography (FMT) system has been in development for the purpose of quantifying the production of Protoporphyrin IX (PPIX) in aminolevulinic acid treated (ALA), Basal Cell Carcinoma (BCC) in vivo. The design couples fiber-based spectral sampling of PPIX fluorescence emission with a high frequency ultrasound imaging system, allowing regionally localized fluorescence intensities to be quantified [1]. The optical data are obtained by sequential excitation of the tissue with a 633nm laser, at four source locations and five parallel detections at each of the five interspersed detection locations. This method of acquisition permits fluorescence detection for both superficial and deep locations in ultrasound field. The optical boundary data, tissue layers segmented from ultrasound image and diffusion theory are used to estimate the fluorescence in tissue layers. To improve the recovery of the fluorescence signal of PPIX, eliminating tissue autofluorescence is of great importance. Here the approach was to utilize measurements which straddled the steep Qband excitation peak of PPIX, via the integration of an additional laser source, exciting at 637 nm; a wavelength with a 2 fold lower PPIX excitation value than 633nm.The auto-fluorescence spectrum acquired from the 637 nm laser is then used to spectrally decouple the fluorescence data and produce an accurate fluorescence emission signal, because the two wavelengths have very similar auto-fluorescence but substantially different PPIX excitation levels. The accuracy of this method, using a single source detector pair setup, is verified through animal tumor model experiments, and the result is compared to different methods of fluorescence signal recovery.

Quantitative photoabsorption and fluorescence spectroscopy of benzene, naphthalene, and some derivatives at 106-295 nm

NASA Technical Reports Server (NTRS)

Suto, Masako; Wang, Xiuyan; Shan, Jun; Lee, L. C.

1992-01-01

Photoabsorption and fluorescence cross sections of benzene, (o-, m-, p-) xylenes, naphthalene, 1-methylnaphthalene, and 2-ethylnaphthalene in the gas phase are measured at 106-295 nm using synchrotron radiation as a light source. Fluorescences are observed from the photoexcitation of benzene and xylenes at 230-280 nm and from naphthalene and its derivatives at 190-295 nm. The absolute fluorescence cross section is determined by calibration with respect to the emission intensity of the NO(A-X) system, for which the fluorescence quantum yield is equal to 1. To cross-check the current calibration method, the quantum yield of the SO2(C-X) system at 220-230 nm was measured since it is about equal to 1. The current quantum-yield data are compared with previously published values measured by different methods.

[Resolving characteristic of CDOM by excitation-emission matrix spectroscopy combined with parallel factor analysis in the seawater of outer Yangtze Estuary in Autumn in 2010].

PubMed

Yan, Li-Hong; Chen, Xue-Jun; Su, Rong-Guo; Han, Xiu-Rong; Zhang, Chuan-Song; Shi, Xiao-Yong

2013-01-01

The distribution and estuarine behavior of fluorescent components of chromophoric dissolved organic matter in the seawater of outer Yangtze Estuary were determined by fluorescence excitation emission matrix spectra combined with parallel factor analysis. Six individual fluorescent components were identified by PARAFAC models, including three terrestrial humic-like components C1 [330 nm/390(430) nm], C2 (390 nm/480 nm), C3 (360 nm/440 nm), marine biological production component C5 (300 nm/400 nm) and protein-like components C4 (290 nm/350 nm) and C6 (275 nm/300 nm). The results indicated that C1, C2, and C3 showed a conservative mixing behavior in the whole estuarine region, especially in high-salinity region. And the fluorescence intensity proportion of C1 and C3 decreased with increase of salinity and fluorescence intensity proportion of C2 kept constant with increase of salinity in the whole estuarine region. While C4 showed conservative mixing behavior in low-salinity region and non-conservative mixing behavior in high-salinity region, and fluorescence intensity proportion of C4 increased with increase of salinity. However, C5 and C6 showed a non-conservative mixing behavior and fluorescence intensity proportion increased with increase of salinity in high-salinity region. Significantly spatial difference was recorded for CDOM absorption coefficient in the coastal region and in the open water areas with the highest value in coastal region and the lowest value in the open water areas. The scope of absorption coefficient and absorption slope was higher in coastal region than that in the open water areas. Significantly positive correlations were found between CDOM absorption coefficient and the fluorescence intensities of C1, C2, C3, and C4, but no significant correlation was found between C5 and C6, suggesting that the river inputs contributed to the coastal areas, while CDOM in the open water areas was affected by terrestrial inputs and phytoplankton degradation.

Emission wavelength tuning of fluorescence by fine structural control of optical metamaterials with Fano resonance

PubMed Central

Moritake, Y.; Kanamori, Y.; Hane, K.

2016-01-01

We demonstrated fine emission wavelength tuning of quantum dot (QD) fluorescence by fine structural control of optical metamaterials with Fano resonance. An asymmetric-double-bar (ADB), which was composed of only two bars with slightly different bar lengths, was used to obtain Fano resonance in the optical region. By changing the short bar length of ADB structures with high dimensional accuracy in the order of 10 nm, resonant wavelengths of Fano resonance were controlled from 1296 to 1416 nm. Fluorescence of QDs embedded in a polymer layer on ADB metamaterials were modified due to coupling to Fano resonance and fine tuning from 1350 to 1376 nm was observed. Wavelength tuning of modified fluorescence was reproduced by analysis using absorption peaks of Fano resonance. Tuning range of modified fluorescence became narrow, which was interpreted by a simple Gaussian model and resulted from comparable FWHM in QD fluorescence and Fano resonant peaks. The results will help the design and fabrication of metamaterial devices with fluorophores such as light sources and biomarkers. PMID:27622503

Large Fluorescence Response by Alcohol from a Bis(benzoxazole)-Zinc(II) Complex: The Role of Excited State Intramolecular Proton Transfer

PubMed Central

Wang, Junfeng; Chu, Qinghui; Liu, Xiumin; Wesdemiotis, Chrys

2013-01-01

The formation of a bis(HBO) anion is known to turn-on the fluorescence to give red emission, via controlling the excited-state intramolecular proton transfer (ESIPT). The poor stability of the formed anion, however, hampered its application. The anion stability is found to be greatly improved by attaching the anion to Zn2+ cation (i.e. forming zinc complex), whose emission is at λem ≈ 550 and 760 nm. Interestingly, addition of methanol to the zinc complex induces a remarkable red fluorescence (λem ≈ 630 nm, ϕfl ≈ 0.8). With the aid of spectroscopic studies (1H NMR, UV-vis, fluorescence, and mass spectra), the structures of the zinc complexes are characterized. The emission species is identified as a dimer-like structure. The study thus reveals an effective fluorescence switching mechanism that could further advance the application of ESIPT-based sensors. PMID:23514312

Ex-vivo UV autofluorescence imaging and fluorescence spectroscopy of atherosclerotic pathology in human aorta

NASA Astrophysics Data System (ADS)

Lewis, William; Williams, Maura; Franco, Walfre

2017-02-01

The aim of our study was to identify fluorescence excitation-emission pairs correlated with atherosclerotic pathology in ex-vivo human aorta. Wide-field images of atherosclerotic human aorta were captured using UV and visible excitation and emission wavelength pairs of several known fluorophores to investigate correspondence with gross pathologic features. Fluorescence spectroscopy and histology were performed on 21 aortic samples. A matrix of Pearson correlation coefficients were determined for the relationship between relevant histologic features and the intensity of emission for 427 wavelength pairs. A multiple linear regression analysis indicated that elastin (370/460 nm) and tryptophan (290/340 nm) fluorescence predicted 58% of the variance in intima thickness (R-squared = 0.588, F(2,18) = 12.8, p=.0003), and 48% of the variance in media thickness (R-squared = 0.483, F(2,18) = 8.42, p=.002), suggesting that endogenous fluorescence intensity at these wavelengths can be utilized for improved pathologic characterization of atherosclerotic plaques.

Implications of the temperature dependence of Nd:YAG spectroscopic values for low temperature laser operation at 946 nm

NASA Astrophysics Data System (ADS)

Yoon, S. J.; Mackenzie, J. I.

2014-05-01

We present our measurements of the key spectroscopic properties over the temperature range of 77 K to 450 K for Nd3+ ions doped in Y3Al5O12 (YAG). From room to liquid nitrogen temperature (LNT), the peak absorption cross section around 808 nm increased by almost 3 times, in conjunction the bandwidth of this absorption line reduced by the same factor. At LNT the peak of the absorption line was blue shifted by 0.25 nm with respect to that at 300 K. The fluorescence spectrum between 850 nm - 1450 nm was measured, from which the emission cross sections for the three main transitions were calculated. One note of particular interest for the dominant emission wavelengths around 1064nm and 1061nm (4F3/2 --> 4I11/2) was the switch in their relative strength below 170K, and at LNT the 1061 nm line has almost twice the cross section as at 1064nm.. The fluorescence and lifetime of the upper laser level (4F3/2) was measured and the effective emission cross section determined by the Fuchtbauer-Ladenburg (F-L) method. The effective emission cross section for 946 nm (R1 --> Z5) increased by more than two times over the 300 K to 77 K range. A numerical fit for the temperature dependent emission cross section at 946 nm and 1064 nm and also calculated absorption coefficient at 808 nm pump diode laser have also obtained from the measured spectroscopic data.

A compact OPO/SFG laser for ultraviolet biological sensing

NASA Astrophysics Data System (ADS)

Tiihonen, Mikael; Pasiskevicius, Valdas; Laurell, Fredrik; Jonsson, Per; Lindgren, Mikael

2004-07-01

A compact parametric oscillator (OPO) with intracavity sum-frequency generation (SFG) to generate 293 nm UV laser irradiation, was developed. The OPO/SFG device was pumped by a 100 Hz Nd:YAG laser (1064 nm) of own design, including subsequent second harmonic generation (SHG) in an external periodically poled KTiOPO4 (KTP) crystal. The whole system could be used to deliver more than 30 μJ laser irradiation per pulse (100 Hz) at 293 nm. The UV laser light was introduced in an optical fiber attached to a sample compartment allowing detection of fluorescence emission using a commercial spectrometer. Aqueous samples containing biomolecules (ovalbumin) or bacteria spores (Bacillus subtilis) were excited by the UV-light at 293 nm resulting in strong fluorescence emission in the range 325 - 600 nm.

Spectral enhancement of leucocrystal violet treated footwear impression evidence in blood.

PubMed

Spence, Lindsay; Asmussen, Gary

2003-03-27

The results presented demonstrate the capacity for spectral enhancement to substantially improve the forensic examination of footwear impressions in blood treated with leucocrystal violet (LCV). The UV-Vis absorption spectra were generated of (i) an aqueous solution of leucocrystal violet, (ii) leucocrystal violet in 3% H(2)O(2), (iii) LCV working solution and (iv) whole blood added to LCV working solution. The resultant fluorescence emission spectra were subsequently generated (lambda(ex)=630nm, lambda(em)=661-900nm). The results indicate that the UV-Vis absorption spectra of an unbuffered solution of whole blood with LCV working solution produces a strong absorbance curve with a maxima at 630nm. Subsequent excitation at this wavelength and generation of the emission spectrum in the fluorescence mode indicates that a solution of whole blood added to LCV working solution is an extremely weak fluorophore. Therefore, to enable an adequate and timely enhancement of blood impression evidence treated with LCV utilising either visible fluorescence or infrared luminescence requires (i) selection of the most appropriate excitation wavelength (lambda(ex)) and emission wavelength (lambda(em)) with extremely narrow band pass filters, which in the absence of substrate matrix interference is excitation at 630nm producing the emission maxima at 665nm and (ii) a visual enhancement system such as a CCD colour IR video camera with image integration.

Impact of oxygen chemistry on the emission and fluorescence spectroscopy of laser ablation plumes

NASA Astrophysics Data System (ADS)

Hartig, K. C.; Brumfield, B. E.; Phillips, M. C.; Harilal, S. S.

2017-09-01

Oxygen present in the ambient gas medium may affect both laser-induced breakdown spectroscopy (LIBS) and laser-induced fluorescence (LIF) emission through a reduction of emission intensity and persistence. In this study, an evaluation is made on the role of oxygen in the ambient environment under atmospheric pressure conditions in LIBS and laser ablation (LA)-LIF emission. To generate plasmas, 1064 nm, 10 ns pulses were focused on an aluminum alloy sample. LIF was performed by frequency scanning a CW laser over the 396.15 nm (3s24s 2S1/2 → 3s23p 2P°3/2) Al I transition. Time-resolved emission and fluorescence signals were recorded to evaluate the variation in emission intensity caused by the presence of oxygen. The oxygen partial pressure (po) in the atmospheric pressure environment using N2 as the makeup gas was varied from 0 to 400 Torr O2. 2D-fluorescence spectroscopy images were obtained for various oxygen concentrations for simultaneous evaluation of the emission and excitation spectral features. Results showed that the presence of oxygen in the ambient environment reduces the persistence of the LIBS and LIF emission through an oxidation process that depletes the density of atomic species within the resulting laser-produced plasma (LPP) plume.

Engineering of mCherry variants with long Stokes shift, red-shifted fluorescence, and low cytotoxicity

PubMed Central

Shen, Yi; Chen, Yingche; Wu, Jiahui; Shaner, Nathan C.; Campbell, Robert E.

2017-01-01

MCherry, the Discosoma sp. mushroom coral-derived monomeric red fluorescent protein (RFP), is a commonly used genetically encoded fluorophore for live cell fluorescence imaging. We have used a combination of protein design and directed evolution to develop mCherry variants with low cytotoxicity to Escherichia coli and altered excitation and emission profiles. These efforts ultimately led to a long Stokes shift (LSS)-mCherry variant (λex = 460 nm and λem = 610 nm) and a red-shifted (RDS)-mCherry variant (λex = 600 nm and λem = 630 nm). These new RFPs provide insight into the influence of the chromophore environment on mCherry’s fluorescence properties, and may serve as templates for the future development of fluorescent probes for live cell imaging. PMID:28241009

Fluorescence properties and energy transfer study of Er3+/Nd3+ doped fluorophosphate glass pumped at 800 and 980 nm for mid-infrared laser applications

NASA Astrophysics Data System (ADS)

Tian, Ying; Xu, Rongrong; Hu, Lili; Zhang, Junjie

2012-04-01

The fluorescence properties of 2.7 μm emission as well as near infrared emissions in Er3+/Nd3+ doped fluorophosphate glasses are investigated under 800 and 980 nm excitation. The fluorescence dynamics and energy transfer processes between Er and Nd ions in different pumping schemes are reported. Three Judd-Ofelt intensity parameters, energy transfer microparameters, and efficiency have been determined using the Judd-Ofelt and Förster-Dexter theories. The calculated energy transfer efficiency of the Er3+:4I13/2 level to the Nd3+:4I15/2 level is as high as 83.91%. The results indicate that Nd3+ may be an efficient sensitizer for Er3+ to obtain mid-infrared emission and the more suitable pumping scheme of 2.7 μm laser applications for Er3+/Nd3+ doped fluorophosphate glass is 980 nm excitation.

Aminoquinolines as fluorescent labels for hydrophilic interaction liquid chromatography of oligosaccharides.

PubMed

Struwe, Weston B; Rudd, Pauline M

2012-08-01

In this study, we investigated the potential of four different aminoquinoline (AQ) compounds as fluorescent labels for glycan analysis using hydrophilic interaction liquid chromatography (HILIC) and fluorescence detection (FLD). We confirmed the optimal excitation and emission wavelengths of 3-AQ and 6-AQ conjugated to glycan standards using three-dimensional fluorescent spectral scanning. The optimal excitation and emission wavelengths for 6-AQ were confirmed at λ(ex)=355 nm and λ(em)=440 nm. We concluded that the optimal wavelengths for 3-AQ were λ(ex)=355 nm and λ(em)=420 nm, which differed considerably from the wavelengths applied in previous reports. HILIC-FLD chromatograms using experimentally determined wavelengths were similar to 2-aminobenzamide controls, but the peak capacity and resolution differed significantly when published 3-AQ λ(ex/em) values were applied. Furthermore, we found that 5-AQ and 8-AQ labeled maltohexaose did not display any fluorescent properties when used as a carbohydrate tag for HPLC analysis. Finally, we applied experimentally determined wavelengths to 3-AQ labeled N-glycans released from human IgG to illustrate changes in retention time as well as to demonstrate that AQ labeling is applicable to complex sample analysis via exoglycosidase sequencing.

Luminescent Fluorene-Based Bis-Pyrazolyl Aniline Ligand for Aluminum Detection.

PubMed

Frazer, Andrew; Morales, Alma R; Woodward, Adam W; Tongwa, Paul; Timofeeva, Tatiana; Belfield, Kevin D

2013-09-29

The design, synthesis, and photophysical properties of a new fluorene-based fluorescent chemosensor, 4-((E)-2-(2-(benzo[d]thiazol-2-yl)-9,9-diethyl-9H-fluoren-7-yl)vinyl)-N,N-bis((3,5-dimethyl-1H-pyrazol-1-yl)methyl)benzenamine (AXF-Al), is described for the detection of Al 3+ . AXF-Al exhibited absorption at 382 nm and strong fluorescence emission at 542 nm (fluorescence quantum yield, Φ F , of 0.80). The capture of Al 3+ by the pyrazolyl aniline receptor resulted in nominal change in the linear absorption (372 nm) but a large hypsochromic shift of 161 nm in the fluorescence spectrum (542 to 433 nm, Φ F  = 0.88), from which Al 3+ was detected both ratiometrically and colorimetrically. The addition of other metal ions, namely Mg 2+ , Ca 2+ , Mn 2+ , Fe 2+ , Co 2+ , Ni 2+ , Cu 2+ , Zn 2+ , Cd 2+ , Hg 2+ and Pb 2+ , produced only minimal changes in the optical properties of this probe. The emission band of this probe was also accessed by two-photon excitation in the near-IR, as two-photon absorption (2PA) is important for potential applications in two-photon fluorescence microscopy (2PFM) imaging. The 2PA cross section of the free fluorenyl ligand AXF-Al was 220 GM at 810 nm and 235 GM at 810 nm for the Al-ligand complex, practically useful properties for 2PFM.

Solvent induced fluorescence enhancement of graphene oxide studied by ultrafast spectroscopy

NASA Astrophysics Data System (ADS)

Zhao, Litao; Chen, Jinquan; He, Xiaoxiao; Yu, Xiantong; Yan, Shujun; Zhang, Sanjun; Pan, Haifeng; Xu, Jianhua

2018-05-01

Femtosecond transient absorption (TA) spectroscopy combined with picosecond time resolved fluorescence (TRF) were used to reveal the fluorescence kinetics of graphene oxide (GO) in water, ethanol and water-ethanol mixtures. Size-independent fluorescence of GO were observed in water, and pH-dependent fluorescence spectra could be fitted well by a triple emission relaxation with peaks around 440 nm, 500 nm, and 590 nm respectively. The results indicate that polycyclic aromatic hydrocarbons (PAHs) linked by oxygen-containing functional groups dominate GO's fluorescence emission. GO's fluorescence quantum yield was measured to be 2.8% in ethanol but 1.2% in water. The three decay components fluorescence decay, as well as the transient absorption dynamics with an offset, confirmed this solvent induced fluorescence enhancement. GO's Raman spectral signals showed that GO in ethanol has a smaller average size of PAHs than that of GO in water. Therefore, besides other enhancement effects reported in literatures, we proposed that solvents could also change the size of PAHs, resulting in a photoluminescence enhancement. Our experimental data demonstrates that GO's quantum yield could be up to 2.8% in water and 8.4% in ethanol and this observation may help ones to improve GO's photoluminescence efficiency as well as its applications in solution.

The fluorescence of a chelating two-photon-absorbing dye is enhanced with the addition of transition metal ions but quenched in the presence of acid

NASA Astrophysics Data System (ADS)

Stewart, David J.; Long, Stephanie L.; Yu, Zhenning; Kannan, Ramamurthi; Mikhailov, Alexandr; Rebane, Aleksander; Tan, Loon-Seng; Haley, Joy E.

2016-09-01

A pseudo-symmetric two-photon absorbing dye (1) containing a central piperazine unit substituted with (benzothiazol-2- yl)-9,9-diethylfluoren-2-yl pendant groups has been synthesized and characterized. The molecule has a two-photonabsorption cross-section of σ2 = 140 GM in tetrahydrofuran at 740 nm and shows significant solvatochromism in the excited-state fluorescence spectra. The emission spectra broaden and the maxima bathochromically shift from 411 nm to 524 nm in n-hexane and acetonitrile, respectively. Moreover, the central piperazine moiety serves as a potential chelation site for ions. Addition of copper(I) hexafluorophosphate and zinc(II) triflate in acetonitrile indicate ground-state complexation with a shift in the emission maximum from 524 nm to 489 nm and 487 nm, respectively. Interestingly, the newly formed Cu and Zn complexes are more strongly emissive than the free dye. Finally, addition of p-toluenesulfonic acid in tetrahydrofuran also blue-shifts the emission maximum, but the intensity is quenched. Due to the photophysical changes induced by addition of metal ions and protons, the dye shows promise as a potential sensor.

Excitation-emission matrices measurements of human cutaneous lesions: tool for fluorescence origin

NASA Astrophysics Data System (ADS)

Zhelyazkova, A.; Borisova, E.; Angelova, L.; Pavlova, E.; Keremedchiev, M.

2013-11-01

The light induced fluorescence (LIF) technique has the potential of providing real-time diagnosis of malignant and premalignant skin tissue; however, human skin is a multilayered and inhomogeneous organ with different optical properties that complicate the analysis of cutaneous fluorescence spectra. In spite of the difficulties related to the detection and analysis of fluorescent data from skin lesions, this technique is among the most widely applied techniques in laboratorial and pre-clinical investigations for early skin neoplasia diagnosis. The important point is to evaluate all sources of intrinsic fluorescence and find any significant alterations distinguishing the normal skin from a cancerous state of the tissue; this would make the autofluorescence signal obtained useful for the development of a non-invasive diagnostic tool for the dermatological practice. Our investigations presented here were based on ex vivo point-by-point measurements of excitation-emission matrices (EEM) from excised tumor lesions and the surrounding skin taken during the daily clinical practice of Queen Jiovanna- ISUL University Hospital, Sofia, the local Ethical Committee's approval having already been obtained. The fluorescence emission was measured between 300 nm and 800 nm using excitation in the 280-440 nm spectral range. In the process of excitation-emission matrices (EEM) measurements we could establish the origin of the autofluorescence and the compounds related by assigning the excitation and emission maxima obtained during the experiments. The EEM were compared for normal human skin, basal cell carcinoma, squamous cell carcinoma, benign nevi and malignant melanoma lesions to obtain information for the most common skin malignancies and their precursors. The main spectral features and the applicability of the technique of autofluorescent spectroscopy of human skin in general as an initial diagnostic tool are discussed as well.

Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf

NASA Astrophysics Data System (ADS)

Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; An Nguyen, Thien; Alfano, Robert R.

2014-06-01

Two-photon (2P) excitation of the second singlet (S) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S2 state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

Deep two-photon microscopic imaging through brain tissue using the second singlet state from fluorescent agent chlorophyll α in spinach leaf.

PubMed

Shi, Lingyan; Rodríguez-Contreras, Adrián; Budansky, Yury; Pu, Yang; Nguyen, Thien An; Alfano, Robert R

2014-06-01

Two-photon (2P) excitation of the second singlet (S₂) state was studied to achieve deep optical microscopic imaging in brain tissue when both the excitation (800 nm) and emission (685 nm) wavelengths lie in the "tissue optical window" (650 to 950 nm). S₂ state technique was used to investigate chlorophyll α (Chl α) fluorescence inside a spinach leaf under a thick layer of freshly sliced rat brain tissue in combination with 2P microscopic imaging. Strong emission at the peak wavelength of 685 nm under the 2P S₂ state of Chl α enabled the imaging depth up to 450 μm through rat brain tissue.

Identification of endogenous flurophores in the layered retina

NASA Astrophysics Data System (ADS)

Xu, Gaixia; Chen, Danni; Sun, Yiwen; Qu, Junle; Lin, Ziyang; Ding, Zhihua; Niu, Hanben

2007-05-01

In this paper, we measured and analyzed the characteristic of endogenous fluorophores in porcine layered retina by using advanced fluorescence spectroscopy and microscopy imaging technology. It was found that there were obvious contrasts corresponding to the different layers of retina, which may be important for fundus disease diagnosis. The retinal pigment epithelium cells exhibited strong autofluorescence with as emission peak of 600+/-10nm when excited with 860-nm light. The emission peak of photoreceptors was at 652+/-5nm, and the emission peak of retinal vessels layer was weak and at 640~700nm, when excited with 488-nm light. Autofluorescence images of three layers of retina were obtained using the same setup. We concluded that the main endogenous fluorophore in PRE was lipofuscin and that in retinal vessels was porphyrin. What's more, the FMHW (full width at half. maximum) of retinal fluorescence spectrum was broad, which suggested that there wasn't only one endogenous fluorophores of tissues excited.

Conversion of just-continuous metallic films to large particulate substrates for metal-enhanced fluorescence

PubMed Central

Aslan, Kadir; Malyn, Stuart N.; Zhang, Yongxia; Geddes, Chris D.

2008-01-01

We report the effects of thermally annealing, non-, just-, and thick continuous silver films for their potential applications in metal-enhanced fluorescence, a near-field concept which can alter the free-space absorption and emissive properties of close-proximity fluorophores (excited states). We have chosen to anneal a noncontinuous particulate film 5 nm thick and two thicker continuous films, 15 and 25 nm thick, respectively. Our results show that the annealing of the 25 nm film has little effect on close-proximity fluorescence when coated with a monolayer of fluorophore-labeled protein. However, the 15 nm continuous film cracks upon annealing, producing large nanoparticles which are ideal for enhancing the fluorescence of close-proximity fluorophores that are indeed difficult to prepare by other wet-chemical deposition processes. The annealing of 5 nm noncontinuous particulate films (a control sample) has little influence on metal-enhanced fluorescence, as expected. PMID:19479004

Conversion of just-continuous metallic films to large particulate substrates for metal-enhanced fluorescence.

PubMed

Aslan, Kadir; Malyn, Stuart N; Zhang, Yongxia; Geddes, Chris D

2008-04-15

We report the effects of thermally annealing, non-, just-, and thick continuous silver films for their potential applications in metal-enhanced fluorescence, a near-field concept which can alter the free-space absorption and emissive properties of close-proximity fluorophores (excited states). We have chosen to anneal a noncontinuous particulate film 5 nm thick and two thicker continuous films, 15 and 25 nm thick, respectively. Our results show that the annealing of the 25 nm film has little effect on close-proximity fluorescence when coated with a monolayer of fluorophore-labeled protein. However, the 15 nm continuous film cracks upon annealing, producing large nanoparticles which are ideal for enhancing the fluorescence of close-proximity fluorophores that are indeed difficult to prepare by other wet-chemical deposition processes. The annealing of 5 nm noncontinuous particulate films (a control sample) has little influence on metal-enhanced fluorescence, as expected.

Fluorescent pH sensor based on Ag@SiO2 core-shell nanoparticle.

PubMed

Bai, Zhenhua; Chen, Rui; Si, Peng; Huang, Youju; Sun, Handong; Kim, Dong-Hwan

2013-06-26

We have demonstrated a novel method for the preparation of a fluorescence-based pH sensor by combining the plasmon resonance band of Ag core and pH sensitive dye (HPTS). A thickness-variable silica shell is placed between Ag core and HPTS dye to achieve the maximum fluorescence enhancement. At the shell thickness of 8 nm, the fluorescence intensity increases 4 and 9 times when the sensor is excited at 405 and 455 nm, respectively. At the same time, the fluorescence intensity shows a good sensitivity toward pH value in the range of 5-9, and the ratio of emission intensity at 513 nm excited at 455 nm to that excited at 405 nm versus the pH value in the range of 5-9 is determined. It is believed that the present pH sensor has the potential for determining pH real time in the biological sample.

Effects of alcohols on fluorescence intensity and color of a discharged-obelin-based biomarker.

PubMed

Alieva, Roza R; Belogurova, Nadezhda V; Petrova, Alena S; Kudryasheva, Nadezhda S

2014-05-01

Photoproteins are responsible for bioluminescence of marine coelenterates; bioluminescent and fluorescent biomarkers based on photoproteins are useful for monitoring of calcium-dependent processes in medical investigations. Here, we present the analysis of intensity and color of light-induced fluorescence of Ca(2+)-discharged photoprotein obelin in the presence of alcohols (ethanol and glycerol). Complex obelin spectra obtained at different concentrations of the alcohols at 350- and 280-nm excitation (corresponding to polypeptide-bound coelenteramide and tryptophan absorption regions) were deconvoluted into Gaussian components; fluorescent intensity and contributions of the components to experimental spectra were analyzed. Five Gaussian components were found in different spectral regions-ultraviolet (tryptophan emission), blue-green (coelenteramide emission), and red (hypothetical indole-coelenteramide exciplex emission). Inhibition coefficients and contributions of the components to experimental fluorescent spectra showed that presence of alcohols increased contributions of ultraviolet, violet, and red components, but decreased contributions of components in the blue-green region. The effects were related to (1) changes of proton transfer efficiency in fluorescent S*1 state of coelenteramide in the obelin active center and (2) formation of indole-coelenteramide exciplex at 280-nm photoexcitation. The data show that variation of fluorescence color and intensity in the presence of alcohols and dependence of emission spectra on excitation wavelength should be considered while applying the discharged obelin as a fluorescence biomarker.

Fluorescence of the various red antenna states in photosystem I complexes from cyanobacteria is affected differently by the redox state of P700.

PubMed

Schlodder, Eberhard; Hussels, Martin; Cetin, Marianne; Karapetyan, Navassard V; Brecht, Marc

2011-11-01

Photosystem I of cyanobacteria contains different spectral pools of chlorophylls called red or long-wavelength chlorophylls that absorb at longer wavelengths than the primary electron donor P700. We measured the fluorescence spectra at the ensemble and the single-molecule level at low temperatures in the presence of oxidized and reduced P700. In accordance with the literature, it was observed that the fluorescence is quenched by P700(+). However, the efficiency of the fluorescence quenching by oxidized P700(+) was found to be extremely different for the various red states in PS I from different cyanobacteria. The emission of the longest-wavelength absorbing antenna state in PS I trimers from Thermosynechococcus elongatus (absorption maximum at 5K: ≈ 719nm; emission maximum at 5K: ≈ 740nm) was found to be strongly quenched by P700(+) similar to the reddest state in PS I trimers from Arthrospira platensis emitting at 760nm at 5K. The fluorescence of these red states is diminished by more than a factor of 10 in the presence of oxidized P700. For the first time, the emission of the reddest states in A. platensis and T. elongatus has been monitored using single-molecule fluorescence techniques. 2011 Elsevier B.V. All rights reserved.

2D and 3D surface photopatterning via laser-promoted homopolymerization of a perfluorophenyl azide-substituted BODIPY.

PubMed

Raffy, Guillaume; Bofinger, Robin; Tron, Arnaud; Guerzo, André Del; McClenaghan, Nathan D; Vincent, Jean-Marc

2017-11-09

An innovative photopatterning process is described that allows, in a single laser-promoted operation, the covalent attachment of a molecule on a surface (2D patterning - xy dimensions) and its photopolymerization to grow micro-/nanostructures with spatial control in a third z-dimension. The surface patterning process, based on nitrene reactivity, was harnessed using the highly fluorescent azide-substituted boron difluoride dipyrromethene (BODIPY) 1 that was prepared in a single synthetic step from the parent pentafluorophenyl BODIPY on reacting with NaN 3 . Using the laser of a fluorescence microscope (375 nm or 532 nm) 1 could be grafted on adapted surfaces and then homopolymerised. In this study we show that using glass coverslips coated with PEG/high density alkyne groups (density of ∼1 × 10 14 per cm 2 ), the patterning process was much more spatially confined than when using PEG only coating. Varying the irradiation time (1 to 15 s) or laser power (0.14-3.53 μW) allowed variation of the amount of deposited BODIPY to afford, in the extreme case, pillars of a height up to 800 nm. AFM and MS studies revealed that the nano/microstructures were formed of particles of photopolymerized 1 having a mean diameter of ca. 30 nm. The emission spectra and fluorescence lifetimes for the patterned structures were measured, revealing a red-shift (from ∼560 nm up to 620 nm) of the maximum emission and a shortening (from ∼6 ns to 0.8 ns) of the fluorescence lifetimes in areas where the density of BODIPY is high. As an application of the patterning process, a figure formed of 136 dots/pillars was prepared. The confocal hyperspectral fluorescence image revealed that the figure is clearly resolved and constituted by highly photoluminescent red dots whose fluorescence intensities and emission color proved to be highly reproducible. SEM and AFM studies showed that the luminescent dots were pillars with a conical shape, an average height of 710 ± 28 nm and a FWHM of 400 ± 20 nm.

Supramolecular nanofiber of pyrene-lactose conjugates and its two-photon fluorescence imaging.

PubMed

Sun, Qian; Zhu, Hong-Yu; Wang, Jun-Fang; Chen, Xiao; Wang, Ke-Rang; Li, Xiao-Liu

2018-04-23

A lactose modified pyrene derivative (Py-Lac) was synthesized, with which novel twisted supramolecular nanofibers in diameter about 20 nm were constructed by self-assembly. The nanofibers showed solid-state fluorescence between 400 nm and 650 nm with the maximum emission at 495 nm. Furthermore, its recognition reaction with PNA lectin was investigated by fluorescence spectra and turbidity assays. It is interesting found that the supramolecular assembly as multivalent glycocluster exhibited unique and selectively binding interactions with PNA lectin with the binding constant of 5.74 × 10 6  M -1 . Moreover, compound Py-Lac showed two-photon fluorescence imaging with Hep G2 cells. Copyright © 2018 Elsevier Inc. All rights reserved.

Preliminary investigation of intrinsic UV fluorescence spectroscopic changes associated with proteolytic digestion of bovine articular cartilage

NASA Astrophysics Data System (ADS)

Lewis, William; Padilla-Martinez, Juan-Pablo; Ortega-Martinez, Antonio; Franco, Walfre

2016-03-01

Degradation and destruction of articular cartilage is the etiology of osteoarthritis (OA), an entity second only to cardiovascular disease as a cause of disability in the United States. Joint mechanics and cartilage biochemistry are believed to play a role in OA; an optical tool to detect structural and chemical changes in articular cartilage might offer benefit for its early detection and treatment. The objective of the present study was to identify the spectral changes in intrinsic ultraviolet (UV) fluorescence of cartilage that occur after proteolytic digestion of cartilage. Bovine articular cartilage samples were incubated in varying concentrations of collagenase ranging from 10ug/mL up to 5mg/mL for 18 hours at 37°C, a model of OA. Pre- and post-incubation measurements were taken of the UV excitation-emission spectrum of each cartilage sample. Mechanical tests were performed to determine the pre- and post-digestion force/displacement ratio associated with indentation of each sample. Spectral changes in intrinsic cartilage fluorescence and stiffness of the cartilage were associated with proteolytic digestion. In particular, changes in the relative intensity of fluorescence peaks associated with pentosidine crosslinks (330 nm excitation, 390 nm emission) and tryptophan (290 nm excitation, 340 nm emission) were found to correlate with different degrees of cartilage digestion and cartilage stiffness. In principle, it may be possible to use UV fluorescence spectral data for early detection of damage to articular cartilage, and as a surrogate measure for cartilage stiffness.

Excitation of sensitized fluorescence of europium and curium in an aqueous solution of thenoyltrifluoroacetone by a nitrogen laser

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

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

1987-03-01

The fluorescence spectrum of trivalent europium in aqueous solutions of thenoyltrifluoroacetone, excited by a nitrogen laser with emission wavelength 337 nm, exhibits bands at 582, 593, 616, 650, and 695 nm. Two bands appear in the fluorescence spectrum of trivalent curium under the same conditions - at 598 and 607 nm. The times of quenching of the fluorescence of the ions of these elements were measured, both in H/sub 2/O medium and in D/sub 2/O. A linear relationship was found between the fluorescence intensity of europium and curium and their concentration in TTA solution. The limit of determination of europiummore » and curium by the fluorescent method with laser excitation using the bands at 615 and 607 nm proved equal to 0.3 and 0.07 ng/ml, respectively.« less

Dual emission fluorescent silver nanoclusters for sensitive detection of the biological coenzyme NAD+/NADH.

PubMed

Yuan, Yufeng; Huang, Kehan; Chang, Mengfang; Qin, Cuifang; Zhang, Sanjun; Pan, Haifeng; Chen, Yan; Xu, Jianhua

2016-02-01

Fluorescent silver nanoclusters (Ag NCs) displaying dual-excitation and dual-emission properties have been developed for the specific detection of NAD(+) (nicotinamide adenine dinucleotide, oxidized form). With the increase of NAD(+) concentrations, the longer wavelength emission (with the peak at 550 nm) was gradually quenched due to the strong interactions between the NAD(+) and Ag NCs, whereas the shorter wavelength emission (peaking at 395 nm) was linearly enhanced. More important, the dual-emission intensity ratio (I395/I550), fitting by a single-exponential decay function, can efficiently detect various NAD(+) levels from 100 to 4000 μM, as well as label NAD(+)/NADH (reduced form of NAD) ratios in the range of 1-50. Copyright © 2015 Elsevier Inc. All rights reserved.

Fluorescent labeling of proteins with amine-specific 1,3,2-(2H)-dioxaborine polymethine dye.

PubMed

Gerasov, Andriy; Shandura, Mykola; Kovtun, Yuriy; Losytskyy, Mykhaylo; Negrutska, Valentyna; Dubey, Igor

2012-01-15

A novel water-soluble amine-reactive dioxaborine trimethine dye was synthesized in a good yield and characterized. The potential of the dye as a specific reagent for protein labeling was demonstrated with bovine serum albumin and lysozyme. Its interaction with proteins was studied by fluorescence spectroscopy and gel electrophoresis. The covalent binding of this almost nonfluorescent dye to proteins results in a 75- to 78-fold increase of its emission intensity accompanied by a red shift of the fluorescence emission maximum by 27 to 45 nm, with fluorescence wavelengths of labeled biomolecules being more than 600 nm. The dye does not require activation for the labeling reaction and can be used in a variety of bioassay applications. Copyright © 2011 Elsevier Inc. All rights reserved.

In vivo native fluorescence spectroscopy and nicotinamide adinine dinucleotide/flavin adenine dinucleotide reduction and oxidation states of oral submucous fibrosis for chemopreventive drug monitoring

NASA Astrophysics Data System (ADS)

Sivabalan, Shanmugam; Vedeswari, C. Ponranjini; Jayachandran, Sadaksharam; Koteeswaran, Dornadula; Pravda, Chidambaranathan; Aruna, Prakasa Rao; Ganesan, Singaravelu

2010-01-01

Native fluorescence spectroscopy has shown potential to characterize and diagnose oral malignancy. We aim at extending the native fluorescence spectroscopy technique to characterize normal and oral submucous fibrosis (OSF) patients under pre- and post-treated conditions, and verify whether this method could also be considered in the monitoring of therapeutic prognosis noninvasively. In this study, 28 normal subjects and 28 clinically proven cases of OSF in the age group of 20 to 40 years are diagnosed using native fluorescence spectroscopy. The OSF patients are given dexamethasone sodium phosphate and hyaluronidase twice a week for 6 weeks, and the therapeutic response is monitored using fluorescence spectroscopy. The fluorescence emission spectra of normal and OSF cases of both pre- and post-treated conditions are recorded in the wavelength region of 350 to 600 nm at an excitation wavelength of 330 nm. The statistical significance is verified using discriminant analysis. The oxidation-reduction ratio of the tissue is also calculated using the fluorescence emission intensities of flavin adenine dinucleotide and nicotinamide adinine dinucleotide at 530 and 440 nm, respectively, and they are compared with conventional physical clinical examinations. This study suggests that native fluorescence spectroscopy could also be extended to OSF diagnosis and therapeutic prognosis.

Perylenetetracarboxylic anhydride as a precursor of fluorescent carbon nanoonion rings.

PubMed

Baldoví, Herme G; Herance, José Raul; Manuel Víctor, Víctor; Alvaro, Mercedes; Garcia, Hermenegildo

2015-08-07

Thermal annealing at 400 °C of perylenetetracarboxylic anhydride in low molecular mass PEG gives rise to the formation of well defined nanoobjects of 2.5 nm height and size distribution from 10 to 65 nm (average 40 nm) after purification of the raw mixture with silicagel chromatography. TEM reveals that the flat nanoobjects are constituted of concentric graphenic rings (0.34 nm interlayer distance). The morphology of the nanoparticles resembles onion rings of nanometric dimensions (nanoonion rings C-NOR). C-NOR particles have an excitation dependent emission with λem from 430 to 570 nm and a maximum emission quantum yield of 0.49. C-NOR particles can be internalized into Hep3B human hepatoma cells as determined by confocal fluorescence microscopy and are remarkably biocompatible affecting slightly cell viability according to the MTT test.

Fluorescence anisotropy of tyrosinate anion using one-, two- and three-photon excitation: tyrosinate anion fluorescence.

PubMed

Kierdaszuk, Borys

2013-03-01

We examined the emission spectra and steady-state anisotropy of tyrosinate anion fluorescence with one-photon (250-310 nm), two-photon (570-620 nm) and three-photon (750-930 nm) excitation. Similar emission spectra of the neutral (pH 7.2) and anionic (pH 13) forms of N-acetyl-L-tyrosinamide (NATyrA) (pKa 10.6) were observed for all modes of excitation, with the maxima at 302 and 352 nm, respectively. Two-photon excitation (2PE) and three-photon excitation (3PE) spectra of the anionic form were the same as that for one-photon excitation (1PE). In contrast, 2PE spectrum from the neutral form showed ~30-nm shift to shorter wavelengths relative to 1PE spectrum (λmax 275 nm) at two-photon energy (550 nm), the latter being overlapped with 3PE spectrum, both at two-photon energy (550 nm). Two-photon cross-sections for NATyrA anion at 565-580 nm were 10 % of that for N-acetyl-L-tryptophanamide (NATrpA), and increased to 90 % at 610 nm, while for the neutral form of NATyrA decreased from 2 % of that for NATrpA at 570 nm to near zero at 585 nm. Surprisingly, the fundamental anisotropy of NATyrA anion in vitrified solution at -60 °C was ~0.05 for 2PE at 610 nm as compared to near 0.3 for 1PE at 305 nm, and wavelength-dependence appears to be a basic feature of its anisotropy. In contrast, the 3PE anisotropy at 900 nm was about 0.5, and 3PE and 1PE anisotropy values appear to be related by the cos(6) θ to cos(2) θ photoselection factor (approx. 10/6) independently of excitation wavelength. Attention is drawn to the possible effect of tyrosinate anions in proteins on their multi-photon induced fluorescence emission and excitation spectra as well as excitation anisotropy spectra.

Renilla luciferase- Aequorea GFP (Ruc-GFP) fusion protein, a novel dual reporter for real-time imaging of gene expression in cell cultures and in live animals.

PubMed

Wang, Y; Yu, Y A; Shabahang, S; Wang, G; Szalay, A A

2002-10-01

Light-emitting reporter proteins play an increasing role in the study of gene expression in vitro and in vivo. Here we present a ruc-gfp fusion gene construct generated by fusing a cDNA for Renilla luciferase (ruc) in-frame with a cDNA encoding the "humanized" GFP (gfp) from Aequorea. A plasmid containing the fusion gene construct was successfully transformed into, and expressed in, mammalian cells. The transformed cells exhibited both Renilla luciferase activity in the presence of coelenterazine and GFP fluorescence upon excitation with UV light. Spectrofluorometry of cells containing the Ruc-GFP fusion protein, in the absence of wavelengths capable of exciting GFP fluorescence but in the presence of the luciferase substrate, coelenterazine, showed an emission spectrum with two peaks at 475 nm and 508 nm. These two peaks correspond to the emission maximum of Renilla luciferase at 475 nm and that of GFP at 508 nm. The peak at 508 nm generated in the presence of coelenterazine alone (without UV excitation) is the result of intramolecular energy transfer from Renilla luciferase to Aequorea GFP. Southern analysis of genomic DNA purified from transformed Chinese hamster ovary (CHO) cells and fluorescence in situ hybridization (FISH) to metaphase chromosomes confirmed the integration of the ruc-gfp fusion gene on a single chromosome. The bifunctional Ruc-GFP fusion protein allows the detection of gene expression at the single-cell level based on green fluorescence, and in a group of cells based on luminescence emission. Furthermore, animal experiments revealed that light emission from the Ruc-GFP fusion protein can be detected externally in the organs or tissues of live animals bearing the gene construct.

Aequorea green fluorescent protein analysis by flow cytometry

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

Ropp, J.D.; Cuthbertson, R.A.; Donahue, C.J.

The isolation and expression of the cDNA for the green fluorescent protein (GFP) from the bioluminescent jellyfish Aequorea victoria has highlighted its potential use as a marker for gene expression in a variety of cell types. The longer wavelength peak (470 nm) of GFP`s bimodal absorption spectrum better matches standard fluorescein filter sets; however, it has a considerably lower amplitude than the major absorption peak at 395. In an effort to increase the sensitivity of GFP with routinely available instrumentation, Heim et al. have generated a GFP mutant (serine-65 to threonine; S65T-GFP) which possesses a single absorption peak centered atmore » 490 nm. We have constructed this mutant in order to determine whether it or wild-type GFP (wt-GFP) afforded greater sensitivity when excited near their respective absorption maxima. Using the conventionally available 488 nm and ultraviolet (UV) laser lines from the argon ion laser as well as the 407 nm line from a krypton ion laser with enhanced violet emission, we were able to closely match the absorption maxima of both the S65T and wild-type forms of Aequorea GFP and analyze differences in fluorescence intensity of transiently transfected 293 cells with flow cytometry. The highest fluorescence signal was observed with 488 nm excitation of S65T-GFP relative to all other laser line/GFP pairs. The wt-GFP fluorescence intensity, in contrast, was significantly higher at 407 nm relative to either 488 nm or UV. These results were consistent with parallel spectrofluorometric analysis of the emission spectrum for wt-GFP and S65T- GFP. The relative contribution of cellular autofluorescence at each wavelength was also investigated and shown to be significantly reduced at 407 nm relative to either UV or 488 nm. 29 refs., 5 figs.« less

Fluorescence spectra and biological activity of aerosolized bacillus spores and MS2 bacteriophage exposed to ozone at different relative humidities in a rotating drum

NASA Astrophysics Data System (ADS)

Ratnesar-Shumate, Shanna; Pan, Yong-Le; Hill, Steven C.; Kinahan, Sean; Corson, Elizabeth; Eshbaugh, Jonathan; Santarpia, Joshua L.

2015-03-01

Biological aerosols (bioaerosols) released into the environment may undergo physical and chemical transformations when exposed to atmospheric constituents such as solar irradiation, reactive oxygenated species, ozone, free radicals, water vapor and pollutants. Aging experiments were performed in a rotating drum chamber subjecting bioaerosols, Bacillus thuringiensis Al Hakam (BtAH) spores and MS2 bacteriophages to ozone at 0 and 150 ppb, and relative humidities (RH) at 10%, 50%, and 80+%. Fluorescence spectra and intensities of the aerosols as a function of time in the reaction chamber were measured with a single particle fluorescence spectrometer (SPFS) and an Ultra-Violet Aerodynamic Particle Sizer® Spectrometer (UV-APS). Losses in biological activity were measured by culture and quantitative polymerase chain reaction (q-PCR) assay. For both types of aerosols the largest change in fluorescence emission was between 280 and 400 nm when excited at 263 nm followed by fluorescence emission between 380 and 700 nm when excited at 351 nm. The fluorescence for both BtAH and MS2 were observed to decrease significantly at high ozone concentration and high RH when excited at 263 nm excitation. The decreases in 263 nm excited fluorescence are indicative of hydrolysis and oxidation of tryptophan in the aerosols. Fluorescence measured with the UV-APS (355-nm excitation) increased with time for both BtAH and MS2 aerosols. A two log loss of MS2 bacteriophage infectivity was observed in the presence of ozone at ~50% and 80% RH when measured by culture and normalized for physical losses by q-PCR. Viability of BtAH spores after exposure could not be measured due to the loss of genomic material during experiments, suggesting degradation of extracelluar DNA attributable to oxidation. The results of these studies indicate that the physical and biological properties of bioaerosols change significantly after exposure to ozone and water vapor.

Fluorescence spectra and biological activity of aerosolized bacillus spores and MS2 bacteriophage exposed to ozone at different relative humidities in a rotating drum

DOE PAGES

Ratnesar-Shumate, Shanna; Pan, Yong-Le; Hill, Steven C.; ...

2015-10-14

Biological aerosols (bioaerosols) released into the environment may undergo physical and chemical transformations when exposed to atmospheric constituents such as solar irradiation, reactive oxygenated species, ozone, free radicals, water vapor and pollutants. Aging experiments were performed in a rotating drum chamber subjecting bioaerosols, Bacillus thuringiensis Al Hakam (BtAH) spores and MS2 bacteriophages to ozone at 0 and 150 ppb, and relative humidities (RH) at 10%, 50%, and 80+%. Fluorescence spectra and intensities of the aerosols as a function of time in the reaction chamber were measured with a single particle fluorescence spectrometer (SPFS) and an Ultra-Violet Aerodynamic Particle Sizer® Spectrometermore » (UV-APS). Losses in biological activity were measured by culture and quantitative polymerase chain reaction (q-PCR) assay. For both types of aerosols the largest change in fluorescence emission was between 280 and 400 nm when excited at 263 nm followed by fluorescence emission between 380 and 700 nm when excited at 351 nm. The fluorescence for both BtAH and MS2 were observed to decrease significantly at high ozone concentration and high RH when excited at 263 nm excitation. The decreases in 263 nm excited fluorescence are indicative of hydrolysis and oxidation of tryptophan in the aerosols. Fluorescence measured with the UV-APS (355-nm excitation) increased with time for both BtAH and MS2 aerosols. A two log loss of MS2 bacteriophage infectivity was observed in the presence of ozone at ~50% and 80% RH when measured by culture and normalized for physical losses by q-PCR. Viability of BtAH spores after exposure could not be measured due to the loss of genomic material during experiments, suggesting degradation of extracelluar DNA attributable to oxidation. The results of these studies indicate that the physical and biological properties of bioaerosols change significantly after exposure to ozone and water vapor.« less

Fluorescence spectra and biological activity of aerosolized bacillus spores and MS2 bacteriophage exposed to ozone at different relative humidities in a rotating drum

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

Ratnesar-Shumate, Shanna; Pan, Yong-Le; Hill, Steven C.

Biological aerosols (bioaerosols) released into the environment may undergo physical and chemical transformations when exposed to atmospheric constituents such as solar irradiation, reactive oxygenated species, ozone, free radicals, water vapor and pollutants. Aging experiments were performed in a rotating drum chamber subjecting bioaerosols, Bacillus thuringiensis Al Hakam (BtAH) spores and MS2 bacteriophages to ozone at 0 and 150 ppb, and relative humidities (RH) at 10%, 50%, and 80+%. Fluorescence spectra and intensities of the aerosols as a function of time in the reaction chamber were measured with a single particle fluorescence spectrometer (SPFS) and an Ultra-Violet Aerodynamic Particle Sizer® Spectrometermore » (UV-APS). Losses in biological activity were measured by culture and quantitative polymerase chain reaction (q-PCR) assay. For both types of aerosols the largest change in fluorescence emission was between 280 and 400 nm when excited at 263 nm followed by fluorescence emission between 380 and 700 nm when excited at 351 nm. The fluorescence for both BtAH and MS2 were observed to decrease significantly at high ozone concentration and high RH when excited at 263 nm excitation. The decreases in 263 nm excited fluorescence are indicative of hydrolysis and oxidation of tryptophan in the aerosols. Fluorescence measured with the UV-APS (355-nm excitation) increased with time for both BtAH and MS2 aerosols. A two log loss of MS2 bacteriophage infectivity was observed in the presence of ozone at ~50% and 80% RH when measured by culture and normalized for physical losses by q-PCR. Viability of BtAH spores after exposure could not be measured due to the loss of genomic material during experiments, suggesting degradation of extracelluar DNA attributable to oxidation. The results of these studies indicate that the physical and biological properties of bioaerosols change significantly after exposure to ozone and water vapor.« less

A photoswitchable orange-to-far-red fluorescent protein, PSmOrange.

PubMed

Subach, Oksana M; Patterson, George H; Ting, Li-Min; Wang, Yarong; Condeelis, John S; Verkhusha, Vladislav V

2011-07-31

We report a photoswitchable monomeric Orange (PSmOrange) protein that is initially orange (excitation, 548 nm; emission, 565 nm) but becomes far-red (excitation, 636 nm; emission, 662 nm) after irradiation with blue-green light. Compared to its parental orange proteins, PSmOrange has greater brightness, faster maturation, higher photoconversion contrast and better photostability. The red-shifted spectra of both forms of PSmOrange enable its simultaneous use with cyan-to-green photoswitchable proteins to study four intracellular populations. Photoconverted PSmOrange has, to our knowledge, the most far-red excitation peak of all GFP-like fluorescent proteins, provides diffraction-limited and super-resolution imaging in the far-red light range, is optimally excited with common red lasers, and can be photoconverted subcutaneously in a mouse. PSmOrange photoswitching occurs via a two-step photo-oxidation process, which causes cleavage of the polypeptide backbone. The far-red fluorescence of photoconverted PSmOrange results from a new chromophore containing N-acylimine with a co-planar carbon-oxygen double bond.

Laser-induced fluorescence detection strategies for sodium atoms and compounds in high-pressure combustors

NASA Technical Reports Server (NTRS)

Weiland, Karen J. R.; Wise, Michael L.; Smith, Gregory P.

1993-01-01

A variety of laser-induced fluorescence schemes were examined experimentally in atmospheric pressure flames to determine their use for sodium atom and salt detection in high-pressure, optically thick environments. Collisional energy transfer plays a large role in fluorescence detection. Optimum sensitivity, at the parts in 10 exp 9 level for a single laser pulse, was obtained with the excitation of the 4p-3s transition at 330 nm and the detection of the 3d-3p fluorescence at 818 nm. Fluorescence loss processes, such as ionization and amplified spontaneous emission, were examined. A new laser-induced atomization/laser-induced fluorescence detection technique was demonstrated for NaOH and NaCl. A 248-nm excimer laser photodissociates the salt molecules present in the seeded flames prior to atom detection by laser-induced fluorescence.

Fluorescence evolution of leachates during treatment processes from two contrasting landfills.

PubMed

Sun, W L; Liu, T T; Cui, F; Ni, J R

2008-10-01

Landfill leachates are composed of a complex mixture of organic matter, including a wide range of potentially fluorescent organic compounds. The fluorescence excitation-emission matrix (FEEM) of leachates during treatment processes is investigated. Particular attention is paid to the fluorescence evolution of leachates during treatment processes. Two typical types of landfill, landfill A (a direct municipal solid waste (MSW) landfill) and landfill B (disposal of bottom ashes from MSW incinerators), in a city in Southern China were selected. The results show that two characteristic and intense excitation-emission peaks located at Ex/Em = 310-330 nm/395-410 nm (peak alpha) and Ex/Em = 250-260 nm/450-460 nm (peak alpha') are observed. As the aromatic chemicals, capable of emitting fluorescence, are more recalcitrant to biodegradation than aliphatic chemicals, enhancement of the dissolved organic carbon normalized fluorescence intensities is demonstrated during treatment processes of leachate A and leachate B. This is confirmed by the variation of ultraviolet absorptivity of leachates at 254 nm. Peak alpha' and peak alpha are attributed to a mixture of xenobiotic organic compounds with low molecular weight and relatively stable aromatic fulvic-like matters with high molecular weight, respectively. Humic substances are more resistant to biodegradation than xenobiotic organic compounds, so a significant reduction in the Ialpha'/Ialpha values (fluorescence intensity ratios of peak alpha' and peak a) of leachate A was observed during treatment processes. However, no evident variation for the Ialpha/Ialpha values of leachate B was found during treatment processes owing to the low concentrations of xenobiotic organic compounds in leachate B after incineration.

Photophysical properties of a laser dye (LD-473) in different solvents

NASA Astrophysics Data System (ADS)

Ibnaouf, K. H.; Alhathlool, R.; Ali, M. K. M.

2018-06-01

In this paper, we investigated the spectroscopic properties the 1, 2, 3, 8-tetrahydrofuran, 2, 3, 8-(LD-473) dissolved in seven types of solvents with different concentrations. The absorption, emission, fluorescence and its quantum yield and Stokes shift of LD-473 were measured. The amplified spontaneous emission (ASE) spectra of LD-473 have been obtained using a transverse laser cavity configuration, where the LD-473 was pumped by laser pulses from the third harmonic of an Nd: YAG laser (355 nm). LD-473 in non-polar solvents exhibits dual ASEs around 445 and 470 nm, whereas the corresponding fluorescence spectrum shows only one peak around 437 nm. The peak at 470 nm is due to the combination of two excited molecules and the solvent between them.

The First Mutant of the Aequorea victoria Green Fluorescent Protein That Forms a Red Chromophore†

PubMed Central

Mishin, Alexander S.; Subach, Fedor V.; Yampolsky, Ilia V.; King, William; Lukyanov, Konstantin A.; Verkhusha, Vladislav V.

2010-01-01

Green fluorescent protein (GFP) from a jellyfish, Aequorea victoria, and its mutants are widely used in biomedical studies as fluorescent markers. In spite of the enormous efforts of academia and industry toward generating its red fluorescent mutants, no GFP variants with emission maximum at more than 529 nm have been developed during the 15 years since its cloning. Here, we used a new strategy of molecular evolution aimed at generating a red-emitting mutant of GFP. As a result, we have succeeded in producing the first GFP mutant that substantially matures to the red-emitting state with excitation and emission maxima at 555 and 585 nm, respectively. A novel, nonoxidative mechanism for formation of the red chromophore in this mutant that includes a dehydration of the Ser65 side chain has been proposed. Model experiments showed that the novel dual-color GFP mutant with green and red emission is suitable for multicolor flow cytometry as an additional color since it is clearly separable from both green and red fluorescent tags. PMID:18366185

Nonnegative constraint analysis of key fluorophores within human breast cancer using native fluorescence spectroscopy excited by selective wavelength of 300 nm

NASA Astrophysics Data System (ADS)

Pu, Yang; Sordillo, Laura A.; Alfano, Robert R.

2015-03-01

Native fluorescence spectroscopy offers an important role in cancer discrimination. It is widely acknowledged that the emission spectrum of tissue is a superposition of spectra of various salient fluorophores. In this study, the native fluorescence spectra of human cancerous and normal breast tissues excited by selected wavelength of 300 nm are used to investigate the key building block fluorophores: tryptophan and reduced nicotinamide adenine dinucleotide (NADH). The basis spectra of these key fluorophores' contribution to the tissue emission spectra are obtained by nonnegative constraint analysis. The emission spectra of human cancerous and normal tissue samples are projected onto the fluorophore spectral subspace. Since previous studies indicate that tryptophan and NADH are key fluorophores related with tumor evolution, it is essential to obtain their information from tissue fluorescence but discard the redundancy. To evaluate the efficacy of for cancer detection, linear discriminant analysis (LDA) classifier is used to evaluate the sensitivity, and specificity. This research demonstrates that the native fluorescence spectroscopy measurements are effective to detect changes of fluorophores' compositions in tissues due to the development of cancer.

One-pot synthesis of fluorescent polysaccharides: adenine grafted agarose and carrageenan.

PubMed

Oza, Mihir D; Prasad, Kamalesh; Siddhanta, A K

2012-08-01

New fluorescent polysaccharides were synthesized by grafting the nucleobase adenine on to the backbones of agarose and κ-carrageenan, which were characterized by FT-IR, (13)C NMR, TGA, XRD, UV, and fluorescence properties. The synthesis involved a rapid water based potassium persulfate (KPS) initiated method under microwave irradiation. The emission spectra of adenine grafted agarose and κ-carrageenan were recorded in aqueous (5×10(-5) M) solution, exhibiting λ(em,max) 347 nm by excitation at 261 nm, affording ca. 30% and 40% enhanced emission intensities, respectively compared to that of pure adenine solution in the same concentration. Similar emission intensity was recorded in the pure adenine solution at its molar equivalent concentrations present in the 5×10(-5) M solution of the agarose and carrageenan grafted products, that is, 3.28×10(-5) M and 4.5×10(-5) M respectively. These fluorescent adenine grafted products may have potential utility in various sensor applications. Copyright © 2012. Published by Elsevier Ltd.

Photoswitchable red fluorescent protein with a large Stokes shift

PubMed Central

Piatkevich, Kiryl D.; English, Brian P.; Malashkevich, Vladimir N.; Xiao, Hui; Almo, Steven C.; Singer, Robert H.; Verkhusha, Vladislav V.

2014-01-01

SUMMARY Subclass of fluorescent proteins, large Stokes shift fluorescent proteins, is characterized by their increased spread between the excitation and emission maxima. Here we report a photoswitchable variant of a red fluorescent protein with a large Stokes shift, PSLSSmKate, which initially exhibits excitation/emission at 445/622 nm, but irradiation with violet light photoswitches PSLSSmKate into a common red form with excitation/emission at 573/621 nm. We characterize spectral, photophysical and biochemical properties of PSLSSmKate in vitro and in mammalian cells, and determine its crystal structure in the large Stokes shift form. Mass-spectrometry, mutagenesis and spectroscopic analysis of PSLSSmKate allow us to propose molecular mechanisms for the large Stokes shift, pH dependence and light-induced chromophore transformation. We demonstrate applicability of PSLSSmKate to superresolution PALM microscopy and protein dynamics in live cells. Given its promising properties, we expect that PSLSSmKate-like phenotype will be further used for photoactivatable imaging and tracking multiple populations of intracellular objects. PMID:25242289

Pharmaceutical analysis in solids using front face fluorescence spectroscopy and multivariate calibration with matrix correction by piecewise direct standardization

NASA Astrophysics Data System (ADS)

Alves, Julio Cesar L.; Poppi, Ronei J.

2013-02-01

This paper reports the application of piecewise direct standardization (PDS) for matrix correction in front face fluorescence spectroscopy of solids when different excipients are used in a pharmaceutical preparation based on a mixture of acetylsalicylic acid (ASA), paracetamol (acetaminophen) and caffeine. As verified in earlier studies, the use of different excipients and their ratio can cause a displacement, change in fluorescence intensity or band profile. To overcome this important drawback, a standardization strategy was adopted to convert all the excitation-emission fluorescence spectra into those used for model development. An excitation-emission matrix (EEM) for which excitation and emission wavelengths ranging from 265 to 405 nm and 300 to 480 nm, respectively, was used. Excellent results were obtained using unfolded partial least squares (U-PLS), with RMSEP values of 8.2 mg/g, 10.9 mg/g and 2.7 mg/g for ASA, paracetamol and caffeine, respectively, and with relative errors lesser than 5% for the three analytes.

Synthesis and Optical Properties of Excited-State Intramolecular Proton Transfer Active π-Conjugated Benzimidazole Compounds: Influence of Structural Rigidification by Ring Fusion.

PubMed

Takagi, Koji; Ito, Kaede; Yamada, Yoshihiro; Nakashima, Takuya; Fukuda, Ryoichi; Ehara, Masahiro; Masu, Hyuma

2017-12-01

Two excited-state intramolecular proton transfer (ESIPT) active benzimidazole derivatives (1 and 2) were synthesized by acid-catalyzed intramolecular cyclization. The steady-state fluorescence spectrum in THF revealed that ring-fused derivative 1 exhibits a dual emission, namely, the major emission was from the K* (keto) form (ESIPT emission) at 515 nm with a large Stokes shift of 11 100 cm -1 and the minor emission was from the E* (enol) form at below 400 nm. In contrast, the normal emission from the E* form was dominant and the fluorescence quantum yield was very low (Φ ∼ 0.002) for nonfused derivative 2. The time-resolved fluorescence spectroscopy of 1 suggested that ESIPT effectively occurs due to the restricted conformational transition to the S 1 -T ICT state, and the averaged radiative and nonradiative decay rate constants were estimated as ⟨k f ⟩ = 0.15 ns -1 and ⟨k nr ⟩ = 0.60 ns -1 , respectively. The fluorescence emission of 1 was influenced by the measurement conditions, such as solvent polarity and basicity, as well as the presence of Lewis base. The ESIPT process and solvatochromic behavior were nicely reproduced by the DFT/TDDFT calculation using the PCM model. In the single-crystal fluorescent spectra, the ESIPT emissions were exclusively observed for both fused and nonfused compounds as a result of hydrogen-bonding interactions.

Picosecond time-resolved fluorescence spectroscopy of K-590 in the bacteriorhodopsin photocycle.

PubMed Central

Atkinson, G H; Blanchard, D; Lemaire, H; Brack, T L; Hayashi, H

1989-01-01

The fluorescence spectrum of a distinct isometric and conformational intermediate formed on the 10(-11) s time scale during the bacteriorhodopsin (BR) photocycle is observed at room temperature using a two laser, pump-probe technique with picosecond time resolution. The BR photocycle is initiated by pulsed (8 ps) excitation at 565 nm, whereas the fluorescence is generated by 4-ps laser pulses at 590 nm. The unstructured fluorescence extends from 650 to 880 nm and appears in the same general spectral region as the fluorescence spectrum assigned to BR-570. The transient fluorescence spectrum can be distinguished from that assigned to BR-570 by a larger emission quantum yield (approximately twice that of BR-570) and by a maximum intensity near 731 nm (shifted 17 nm to higher energy from the maximum of the BR-570 fluorescence spectrum). The fluorescence spectrum of BR-570 only is measured with low energy, picosecond pulsed excitation at 590 nm and is in good agreement with recent data in the literature. The assignment of the transient fluorescence spectrum to the K-590 intermediate is based on its appearance at time delays longer than 40 ps. The K-590 fluorescence spectrum remains unchanged over the entire 40-100-ps interval. The relevance of these fluorescence data with respect to the molecular mechanism used to model the primary processes in the BR photocycle also is discussed. PMID:2713439

Effect of fluorescent particle size on the modulation efficiency of ultrasound-modulated fluorescence.

PubMed

Liu, Yuan; Yuan, Baohong; Vignola, Joseph

2012-01-01

To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes.

Effect of fluorescent particle size on the modulation efficiency of ultrasound-modulated fluorescence

PubMed Central

Liu, Yuan; Yuan, Baohong; Vignola, Joseph

2013-01-01

To investigate whether the size of fluorescent particles affects the modulation efficiency of ultrasound-modulated fluorescence (UMF), we measured UMF and DC (direct current) signals of the fluorescence emission from four different sized fluorescent particles: (1) three carboxylate-modified fluorescent microspheres (FM) with diameters of 20 nm, 200 nm, and 1.0 µm and (2) streptavidin-conjugated Alexa Fluor 647 with a diameter of approximately 5 nm. The UMF and DC signals were simultaneously measured using a broadband lock-in amplifier and a narrowband amplifier, respectively. The ratio of the UMF strength to the DC signal strength is defined as the modulation efficiency. This modulation efficiency was then used to evaluate the effects of fluorophore size and concentration. Results show that the modulation efficiency was improved by approximately a factor of two when the size of the fluorescent particles is increased from 5 nm to 1 µm. In addition, the linear relationship between the UMF strength and ultrasound pressure (observed in our previous study) were maintained regardless of the fluorescent particle sizes. PMID:24179476

Highly selective and sensitive nanoprobes for cyanide based on gold nanoclusters with red fluorescence emission

NASA Astrophysics Data System (ADS)

Zhang, Guomei; Qiao, Yunyun; Xu, Ting; Zhang, Caihong; Zhang, Yan; Shi, Lihong; Shuang, Shaomin; Dong, Chuan

2015-07-01

We report a novel and environmentally friendly fluorescent probe for detecting the cyanide ion (CN-) using l-amino acid oxidase (LAAOx)-protected Au nanoclusters (LAAOx@AuNCs) with red emission. The fluorescence-based sensing behaviour of LAAOx@AuNCs towards anions was investigated in buffered aqueous media. Among the anions studied, CN- was found to effectively quench the fluorescence emission of AuNCs based on CN- induced Au core decomposition. Excellent sensitivity and selectivity toward the detection of CN- in aqueous solution were observed. The CN- detection limit was determined to be approximately 180 nM, which is 15 times lower than the maximum level (2700 nM) of CN- in drinking water permitted by the World Health Organization (WHO). A linear relationship between the fluorescence intensity and CN- concentration was observed in two ranges of CN- concentration, including 3.2 × 10-6 to 3.4 × 10-5 mol L-1 and 3.81 × 10-5 to 1.04 × 10-4 mol L-1. The high sensitivity and selectivity to CN- among the 17 types of anions make the AuNCs good candidates for use in fluorescent nanoprobes of CN-.

Authentication of the botanical and geographical origin of honey by front-face fluorescence spectroscopy.

PubMed

Ruoff, Kaspar; Luginbühl, Werner; Künzli, Raphael; Bogdanov, Stefan; Bosset, Jacques Olivier; von der Ohe, Katharina; von der Ohe, Werner; Amado, Renato

2006-09-06

Front-face fluorescence spectroscopy, directly applied on honey samples, was used for the authentication of 11 unifloral and polyfloral honey types (n = 371 samples) previously classified using traditional methods such as chemical, pollen, and sensory analysis. Excitation spectra (220-400 nm) were recorded with the emission measured at 420 nm. In addition, emission spectra were recorded between 290 and 500 nm (excitation at 270 nm) as well as between 330 and 550 nm (excitation at 310 nm). A total of four different spectral data sets were considered for data analysis. Chemometric evaluation of the spectra included principal component analysis and linear discriminant analysis; the error rates of the discriminant models were calculated by using Bayes' theorem. They ranged from <0.1% (polyfloral and chestnut honeys) to 9.9% (fir honeydew honey) by using single spectral data sets and from <0.1% (metcalfa honeydew, polyfloral, and chestnut honeys) to 7.5% (lime honey) by combining two data sets. This study indicates that front-face fluorescence spectroscopy is a promising technique for the authentication of the botanical origin of honey and may also be useful for the determination of the geographical origin within the same unifloral honey type.

The role of ultraviolet-A reflectance and ultraviolet-A induced fluorescence in the appearance of budgerigar plumage: insights from spectrofluorometry and reflectance spectrophotometry.

PubMed Central

Pearn, Sophie M; Bennett, Andrew T D; Cuthill, Innes C

2003-01-01

Fluorescence has so far been found in 52 parrot species when illuminated with ultraviolet-A (UVA) 'black' lamps, and two attempts have been made to determine whether such fluorescence plays any role in sexual signalling. However, the contribution of the reflectance versus fluorescence to the total radiance from feathers, even in the most studied species to date (budgerigars), is unclear. Nor has the plumage of this study species been systematically assessed to determine the distribution of fluorescent patches. We therefore used spectrofluorometry to determine which areas of budgerigars fluoresce and the excitation and emission spectra involved; this is the first time that such a technique has been applied to avian plumage. We found that both the yellow crown and (normally hidden) white downy chest feathers exhibit strong UVA-induced fluorescence, with peak emissions at 527 nm and 436 nm, respectively. Conversely, the bright-green chest and dark-blue tail feathers do not fluoresce. When comparing reflectance spectra (400-700 nm) from the yellow crown using illuminants with a proportion of UVA comparable to daylight, and illuminants with all UVA removed, no measurable difference resulting from fluorescence was found. This suggests that under normal daylight the contribution of fluorescence to radiance is probably trivial. Furthermore, these spectra revealed that males had fluorescent crowns with substantially higher reflectance than those of females, in both the UV waveband and at longer wavelengths. Reflectance spectrophotometry was also performed on a number of live wild-type male budgerigars to investigate the chromatic contrast between the different plumage areas. This showed that many plumage regions are highly UV-reflective. Overall our results suggest that rapid surveys using UVA black lamps may overestimate the contribution of fluorescence to plumage coloration, and that any signalling role of fluorescence emissions, at least from the yellow crown of budgerigars, may not be as important as previously thought. PMID:12737665

Preliminary study of diagnostic spectroscopic imaging for nasopharyngeal carcinoma

NASA Astrophysics Data System (ADS)

Li, Buhong; Xie, Shusen; Zhang, Xiaodong; Li, Depin

2003-12-01

The optical biopsy system for nasopharyngeal carcinoma based on the technique of laser-induced exogenous fluorescence has been successful developed. Ar+ laser was selected as the excitation light source based on the measurement of the Emission-Excitation Matrix of Hematoporphyrin Monomethyl Ether. Tissue-simulating optical phantoms diluted with different concentration of HMME were used to simulated nasopharyngeal carcinoma lesions in the performance test for the drug-fluorescence optical biopsy system, especially for the comparison of fluorescence image contrast between the excitation wavelength of 488nm and 514.5nm, respectively. Experimental results show that the fluorescence image contrast of simulated nasopharyngeal carcinoma lesions excited by the light at the wavelength of 488nm is about three fold higher than that at 514.5nm, and the sensitivity and resolution of the fluorescence and reflection twilight image can satisfy the needs for clinical diagnosis and localization.

Coherent stimulated light emission (lasing) in covalently linked chlorophyll dimers

PubMed Central

Hindman, James C.; Kugel, Roger; Wasielewski, Michael R.; Katz, Joseph J.

1978-01-01

The covalently linked chlorophyll a dimer exhibits remarkably different properties in the folded and open configurations. In the folded configuration the absorption maximum is at 695 nm and the fluorescence maximum is at 730 nm. Laser output at 733 and 735 nm is obtained for solutions in wet benzene and 0.1 M ethanol/toluene, respectively. Measurements of fluorescence lineshapes, made with a transverse excited atmospheric (TEA) nitrogen laser for excitation, show the lifetime shortening associated with stimulated emission resulting from appreciable concentrations of molecules in S1 excited states. In contrast, the open dimer has absorption and fluorescence spectra essentially the same as those of chlorophyll a monomer. Unlike either the folded dimer or chlorophyll a monomer, the open dimer shows no laser emission or fluorescene lifetime shortening. It does not appear that the behavior of the open dimer can be explained in terms of excimer or triplet formation or by nonradiative decay processes. It is suggested that absorption of the exciting radiation by S1, leading to the formation of an exciplex or charge transfer state, may be involved. Significantly, no large changes in fluorescence quantum yield or fluorescence lifetime are observed for these dimers as compared to monomer chlorophyll. This suggests that concentration quenching and lifetime shortening in condensed chlorophyll systems involve more than the simple proximity of two chlorophyll molecules. Images PMID:16592524

UV plasmonic enhancement through three dimensional nano-cavity antenna array in aluminum

NASA Astrophysics Data System (ADS)

Mao, Jieying; Stevenson, Peter; Montanaric, Danielle; Wang, Yunshan; Shumaker-Parry, Jennifer S.; Harris, Joel M.; Blair, Steve

2017-08-01

Metallic nanostructure can enhance fluorescence through excited surface plasmons which increase the local field as well as improve its quantum efficiency. When coupling to cavity resonance with proper gap dimension, gap hot spots can be generated to interact with fluorescence at their excitation/emission region in UV. A 3D nano-cavity antenna array in Aluminum has been conducted to generate local hot spot resonant at fluorescence emission resonance. Giant field enhancement has been achieved through coupling fundamental resonance modes of nanocavity into surface plasmons polaritons (SPPs). In this work, two distinct plasmonic structure of 3D resonant cavity nanoantenna has been studied and its plasmonic response has been scaled down to the UV regime through finite-difference-time-domain (FDTD) method. Two different strategies for antenna fabrication will be conducted to obtain D-coupled Dots-on-Pillar Antenna array (D2PA) through Focus Ion Beam (FIB) and Cap- Hole Pair Antenna array (CHPA) through nanosphere template lithography (NTL). With proper optimization of the structures, D2PA and CHPA square array with 280nm pitch have achieved distinct enhancement at fluorophore emission wavelength 350nm and excitation wavelength 280nm simultaneously. Maximum field enhancement can reach 20 and 65 fold in the gap of D2PA and CHPA when light incident from substrate, which is expected to greatly enhance fluorescent quantum efficiency that will be confirmed in fluorescence lifetime measurement.

Bowel perforation detection using metabolic fluorescent chlorophylls

NASA Astrophysics Data System (ADS)

Han, Jung Hyun; Jo, Young Goun; Kim, Jung Chul; Choi, Sujeong; Kang, Hoonsoo; Kim, Yong-Chul; Hwang, In-Wook

2016-03-01

Thus far, there have been tries of detection of disease using fluorescent materials. We introduce the chlorophyll derivatives from food plants, which have longer-wavelength emissions (at >650 nm) than those of fluorescence of tissues and organs, for detection of bowel perforation. To figure out the possibility of fluorescence spectroscopy as a monitoring sensor of bowel perforation, fluorescence from organs of rodent models, intestinal and peritoneal fluids of rodent models and human were analyzed. In IVIS fluorescence image of rodent abdominal organ, visualization of perforated area only was possible when threshold of image is extremely finely controlled. Generally, both perforated area of bowel and normal bowel which filled with large amount of chlorophyll derivatives were visualized with fluorescence. The fluorescence from chlorophyll derivatives penetrated through the normal bowel wall makes difficult to distinguish perforation area from normal bowel with direct visualization of fluorescence. However, intestinal fluids containing chlorophyll derivatives from food contents can leak from perforation sites in situation of bowel perforation. It may show brighter and longer-wavelength regime emissions of chlorophyll derivatives than those of pure peritoneal fluid or bioorgans. Peritoneal fluid mixed with intestinal fluids show much brighter emissions in longer wavelength (at>650 nm) than those of pure peritoneal fluid. In addition, irrigation fluid, which is used for the cleansing of organ and peritoneal cavity, made of mixed intestinal and peritoneal fluid diluted with physiologic saline also can be monitored bowel perforation during surgery.

Towards metabolic mapping of the human retina.

PubMed

Schweitzer, D; Schenke, S; Hammer, M; Schweitzer, F; Jentsch, S; Birckner, E; Becker, W; Bergmann, A

2007-05-01

Functional alterations are first signs of a starting pathological process. A device that measures parameter for the characterization of the metabolism at the human eye-ground would be a helpful tool for early diagnostics in stages when alterations are yet reversible. Measurements of blood flow and of oxygen saturation are necessary but not sufficient. The new technique of auto-fluorescence lifetime measurement (FLIM) opens in combination with selected excitation and emission ranges the possibility for metabolic mapping. FLIM not only adds an additional discrimination parameter to distinguish different fluorophores but also resolves different quenching states of the same fluorophore. Because of its high sensitivity and high temporal resolution, its capability to resolve multi-exponential decay functions, and its easy combination with laser scanner ophthalmoscopy, multi-dimensional time-correlated single photon counting was used for fundus imaging. An optimized set up for in vivo lifetime measurements at the human eye-ground will be explained. In this, the fundus fluorescence is excited at 446 or 468 nm and the time-resolved autofluorescence is detected in two spectral ranges between 510 and 560 nm as well as between 560 and 700 nm simultaneously. Exciting the fundus at 446 nm, several fluorescence maxima of lifetime t1 were detected between 100 and 220 ps in lifetime histograms of 40 degrees fundus images. In contrast, excitation at 468 nm results in a single maximum of lifetime t1 = 190 +/- 16 ps. Several fundus layers contribute to the fluorescence intensity in the short-wave emission range 510-560 nm. In contrast, the fluorescence intensity in the long-wave emission range between 560 and 700 nm is dominated by the fluorescence of lipofuscin in the retinal pigment epithelium. Comparing the lateral distribution of parameters of a tri-exponential model function in lifetime images of the fundus with the layered anatomical fundus structure, the shortest component (t1 = 190 ps) originates from the retinal pigment epithelium and the second lifetime (t2 = 1,000 ps) from the neural retina. The lifetime t3 approximately 5.5 ns might be influenced by the long decay of the fluorescence in the crystalline lens. In vitro analysis of the spectral properties of expected fluorophores under the condition of the living eye lightens the interpretation of in vivo measurements. Taking into account the transmission of the ocular media, the excitation of NADH is unlikely at the fundus. Copyright 2007 Wiley-Liss, Inc.

Emission cross sections of excited fragments produced by electron impact on BCl3

NASA Astrophysics Data System (ADS)

Tokue, Ikuo; Kudo, Mikiko; Kusakabe, Masanobu; Honda, Tomohisa; Ito, Yoshio

1992-06-01

Emission spectra in the 190-600 nm region produced by electron impact on BCl3 have been studied up to 110 eV. Emission cross sections of the B(2s2p2 2D-2p 2P0) and B(3s 2S-2p 2P0) lines and the BCl(A 1Π-X 1Σ+) band are evaluated to be 4.9±1.0, 4.5±0.7, and (1.9±0.3)×10-18 cm2, respectively, at 100 eV. Formation cross sections of these species have been determined from the analysis of their fluorescence decaying curves. Two continuous emissions observed in the 230-380 and 400-580 nm regions are attributed to the BCl*2 band. The fluorescence lifetime of BCl*2 in the 300-342 nm region is obtained to be 1.65±0.2 μs, which is nearly independent of the wavelength.

Melanin-originated carbonaceous dots for triple negative breast cancer diagnosis by fluorescence and photoacoustic dual-mode imaging.

PubMed

Xiao, Wei; Li, Yuan; Hu, Chuan; Huang, Yuan; He, Qin; Gao, Huile

2017-07-01

Carbonaceous dots exhibit increasing applications in diagnosis and drug delivery due to excellent photostability and biocompatibility properties. However, relative short excitation and emission of melanin carbonaceous dots (MCDs) limit the applicability in fluorescence bioimaging. Furthermore, the generally poor spatial resolution of fluorescence imaging limits potential in vivo applications. Due to a variety of beneficial properties, in this study, MCDs were prepared exhibiting great potential in fluorescence and photoacoustic dual-mode bioimaging. The MCDs exhibited a long excitation peak at 615nm and emission peak at 650nm, further highlighting the applicability in fluorescence imaging, while the absorbance peak at 633nm renders MCDs suitable for photoacoustic imaging. In vivo, the photoacoustic signal of MCDs was linearly correlated with the concentration of MCDs. Moreover, the MCDs were shown to be taken up into triple negative breast cancer cell line 4T1 in both a time- and concentration-dependent manner. In vivo fluorescence and photoacoustic imaging of subcutaneous 4T1 tumor demonstrated that MCDs could passively target triple negative breast cancer tissue by enhanced permeability and retention effects and may therefore be used for tumor dual-mode imaging. Furthermore, fluorescence distribution in tissue slices suggested that MCDs may distribute in 4T1 tumor with high efficacy. In conclusion, the MCDs studied offer potential application in fluorescence and photoacoustic dual-mode imaging. Copyright © 2017 Elsevier Inc. All rights reserved.

Near-Infrared Phosphorus-Substituted Rhodamine with Emission Wavelength above 700 nm for Bioimaging.

PubMed

Chai, Xiaoyun; Cui, Xiaoyan; Wang, Baogang; Yang, Fan; Cai, Yi; Wu, Qiuye; Wang, Ting

2015-11-16

Phosphorus has been successfully fused into a classic rhodamine framework, in which it replaces the bridging oxygen atom to give a series of phosphorus-substituted rhodamines (PRs). Because of the electron-accepting properties of the phosphorus moiety, which is due to effective σ*-π* interactions and strengthened by the inductivity of phosphine oxide, PR exhibits extraordinary long-wavelength fluorescence emission, elongating to the region above 700 nm, with bathochromic shifts of 140 and 40 nm relative to rhodamine and silicon-substituted rhodamine, respectively. Other advantageous properties of the rhodamine family, including high molar extinction coefficient, considerable quantum efficiency, high water solubility, pH-independent emission, great tolerance to photobleaching, and low cytotoxicity, stay intact in PR. Given these excellent properties, PR is desirable for NIR-fluorescence imaging in vivo. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

A comparison study of different excitation wavelengths to determine the relative content of key biomolecules in breast cancer and breast normal tissue

NASA Astrophysics Data System (ADS)

Sordillo, Laura A.; Sordillo, Peter P.; Budansky, Yury; Pu, Yang; Alfano, R. R.

2015-03-01

Fluorescence profiles from breast cancer and breast normal tissue samples with excitation wavelengths at 280 nm and 340 nm were obtained using the conventional LS-50 Perkin-Elmer spectrometer. Fluorescence ratios from these tissue samples, demonstrated by emission peaks at 340 nm, 440 nm and 460 nm and likely representing tryptophan and NADH, show increased relative content of tryptophan in malignant samples. Double ratio (DR) techniques were used to measure the severity of disease. The single excitation double ratio (Single-DR) method utilizes the emission intensity peaks from the spectrum acquired using a single excitation of 280 nm; while the dual excitation double ratio (dual-DR) method utilizes the emission intensity peaks from the spectra acquired using an excitation of 280 nm and 340 nm. Single-DR and dual-DR from 13 patients with breast carcinoma were compared in terms of their efficiency to distinguish high from low/intermediate tumors. Similar results were found with both methods. Results suggest that dual excitation wavelengths may be as effective as single excitation wavelength in calculating the relative content of biomolecules in breast cancer tissue, as well as for the assessment of the malignant potential of these tumors.

Influence of the CdSe quantum dots concentration on the amplified spontaneous emission from the conjugated polymer (MEH-PPV) in solution

NASA Astrophysics Data System (ADS)

Ibnaouf, K. H.

2015-04-01

The spectral properties of a conjugated polymer poly [2-methoxy-5-(2-ethylhexyloxy)-1, 4-phenylenevinylene] (MEH-PPV) in benzene have been studied intensively. The fluorescence spectra for MEH-PPV, under low concentrations, have shown two peaks around 560 nm and 600 nm, which could be attributed to the monomer and excimer states respectively. In our earlier communication, we had shown that MEH-PPV alone could produce amplified spontaneous emission (ASE) only in its excimeric state (600 nm). The spectral properties of 5 nm size of CdSe (core) quantum dots have been investigated. The fluorescence spectra of CdSe core in benzene showed only one band at 590 nm. Mixtures made of MEH-PPV and CdSe (core) quantum dots have been utilized for studying the amplified spontaneous emission characteristics (ASE) in an organic solution under laser excitation. When the mixture was pumped by the third harmonic of Nd:YAG (355 nm), we observed two ASE peaks; one at 575 nm and another at 595 nm. These ASE peaks could arise from the monomer and excimer states of MEH-PPV. This is perhaps the first report on the influence of quantum dots on the laser from the conjugated polymer MEH-PPV, in liquid solution.

Time-resolved delayed luminescence image microscopy using an europium ion chelate complex.

PubMed Central

Marriott, G.; Heidecker, M.; Diamandis, E. P.; Yan-Marriott, Y.

1994-01-01

Improvements and extended applications of time-resolved delayed luminescence imaging microscopy (TR-DLIM) in cell biology are described. The emission properties of europium ion complexed to a fluorescent chelating group capable of labeling proteins are exploited to provide high contrast images of biotin labeled ligands through detection of the delayed emission. The streptavidin-based macromolecular complex (SBMC) employs streptavidin cross-linked to thyroglobulin multiply labeled with the europium-fluorescent chelate. The fluorescent chelate is efficiently excited with 340-nm light, after which it sensitizes europium ion emission at 612 nm hundreds of microseconds later. The SBMC complex has a high quantum yield orders of magnitude higher than that of eosin, a commonly used delayed luminescent probe, and can be readily seen by the naked eye, even in specimens double-labeled with prompt fluorescent probes. Unlike triplet-state phosphorescent probes, sensitized europium ion emission is insensitive to photobleaching and quenching by molecular oxygen; these properties have been exploited to obtain delayed luminescence images of living cells in aerated medium thus complementing imaging studies using prompt fluorescent probes. Since TR-DLIM has the unique property of rejecting enormous signals that originate from scattered light, autofluorescence, and prompt fluorescence it has been possible to resolve double emission images of living amoeba cells containing an intensely stained lucifer yellow in pinocytosed vesicles and membrane surface-bound SBMC-labeled biotinylated concanavalin A. Images of fixed cells represented in terms of the time decay of the sensitized emission show the lifetime of the europium ion emission is sensitive to the environment in which it is found. Through the coupling of SBMC to streptavidin,a plethora of biotin-based tracer molecules are available for immunocytochemical studies. Images FIGURE 1 FIGURE 2 FIGURE 3 FIGURE 4 FIGURE 5 FIGURE 6 FIGURE 7 PMID:7811952

Photophysical parameters and fluorescence quenching of 7-diethylaminocoumarin (DEAC) laser dye

NASA Astrophysics Data System (ADS)

El-Mossalamy, E. H.; Obaid, A. Y.; El-Daly, S. A.

2011-10-01

The optical properties including electronic absorption spectrum, emission spectrum, fluorescence quantum yield, and dipole moment of electronic transition of 7-diethylaminocoumarin (DEAC) laser dye have been measured in different solvents. Both electronic absorption and fluorescence spectra are red shifted as the polarity of the medium increases, indicating that the dipole moment of molecule increases on excitation. The fluorescence quantum yield of DEAC decreases as the polarity of solvent increases, a result of the role of solvent polarity in stabilization of the twisting of the intramolecular charge transfer (TICT) in excited state, which is a non-emissive state, as well as hydrogen bonding with the hetero-atom of dye. The emission spectrum of DEAC has also been measured in cationic (CTAC) and anionic (SDS) micelles, the intensity increases as the concentration of surfactant increases, and an abrupt change in emission intensity is observed at critical micelle concentration (CMC) of surfactant. 2×10 -3 mol dm -3 of DEAC gives laser emission in the blue region on pumping with nitrogen laser ( λex=337.1 nm). The laser parameters such as tuning range, gain coefficient ( α), emission cross section ( σe), and half-life energy have been calculated in different solvents, namely acetone, dioxane , ethanol, and dimethyforamide (DMF). The photoreactivity of DEAC has been studied in CCl 4 at a wavelength of 366 nm. The values of photochemical yield ( ϕc) and rate constant ( k) are determined. The interaction of organic acceptors such as picric acid (PA), tetracyanoethylene (TCNE), and 7,7,8,8-tetracynoquinonedimethane (TCNQ) with DEAC is also studied using fluorescence measurements in acetonitrile (CH 3CN); from fluorescence quenching study we assume the possible electron transfer from excited donor DEAC to organic acceptor forming non-emissive exciplex.

Simultaneous detection of Escherichia coli O157:H7 and Salmonella Typhimurium using quantum dots as fluorescence labels.

PubMed

Yang, Liju; Li, Yanbin

2006-03-01

In this study, we explored the use of semiconductor quantum dots (QDs) as fluorescence labels in immunoassays for simultaneous detection of two species of foodborne pathogenic bacteria, Escherichia coli O157:H7 and Salmonella Typhimurium. QDs with different sizes can be excited with a single wavelength of light, resulting in different emission peaks that can be measured simultaneously. Highly fluorescent semiconductor quantum dots with different emission wavelengths (525 nm and 705 nm) were conjugated to anti-E. coli O157 and anti-Salmonella antibodies, respectively. Target bacteria were separated from samples by using specific antibody coated magnetic beads. The bead-cell complexes reacted with QD-antibody conjugates to form bead-cell-QD complexes. Fluorescent microscopic images of QD labeled E. coli and Salmonella cells demonstrated that QD-antibody conjugates could evenly and completely attach to the surface of bacterial cells, indicating that the conjugated QD molecules still retain their effective fluorescence, while the conjugated antibody molecules remain active and are able to recognize their specific target bacteria in a complex mixture. The intensities of fluorescence emission peaks at 525 nm and 705 nm of the final complexes were measured for quantitative detection of E. coli O157:H7 and S. Typhimurium simultaneously. The fluorescence intensity (FI) as a function of cell number (N) was found for Salmonella and E. coli, respectively. The regression models can be expressed as: FI = 60.6 log N- 250.9 with R(2) = 0.97 for S. Typhimurium, and FI = 77.8 log N- 245.2 with R(2) = 0.91 for E. coli O157:H7 in the range of cell numbers from 10(4) to 10(7) cfu ml(-1). The detection limit of this method was 10(4) cfu ml(-1). The detection could be completed within 2 hours. The principle of this method could be extended to detect multiple species of bacteria (3-4 species) simultaneously, depending on the availability of each type of QD-antibody conjugates with a unique emission peak and the antibody coated magnetic beads specific to each species of bacteria.

Fabrication et caracterisation de cristaux photoniques pour exaltation de fluorescence

NASA Astrophysics Data System (ADS)

Gascon, Annabelle

2011-12-01

In today's world, there is a pressing need for point-of-care molecular analysis that is fast, inexpensive and transportable. Lab-on-a- chips are designed to fulfill that need. They are micro-electromechanical systems (MEMS), fabricated with microelectronic techniques, that use the analytes physical properties to detect their presence in liquid samples. This detection can be performed by attaching the analyte to quantum dots. These quantum dots are semiconducting nanoparticles with narrow fluorescence band. In our project, we use a tuneable system with a two-slab photonic crystal that serves as a tuneable optical filter, detecting the presence and wavelength of these quantum dots. Photonic crystals are dielectrics with a variable refractive index, with a period near the visible light wavelength. They are called photonic crystals because they have a photonic band gap just as atomic crystals, periodic structure of atoms, have an electronic band gap. They are photonic because photons instead of electrons propagate through them. They can also enhance fluorescence from quantum dots at the photonic crystals guided resonance wavelength. My project objectives are to: (1) Fabricate two-slab photonic crystal, (2) Characterize photonic crystals, (3) Place quantum dots on photonic crystals, (4) Measure fluorescence enhancement. The device made during this project consists of a silicon wafer on which were deposited a 200 nm silicon nitride layer, then a 200 nm silicon dioxide layer and finally another 200 nm silicon nitride layer. An electron-beam lithography defines the photonic crystals and the MEMS. The photonic crystals are square lattices of holes 180 nm in diameter, at a period of 460 nm, etched through the two silicon nitride slabs. The two slabs are etched in a single step of Reactive Ion Etching (RIE). Then, the silicon under the photonic crystal is etched from the backside up to the nitride by deep-RIE. Finally, the oxide layer is removed in order to completely suspend the two-slab photonic crystal. The M EMS can change the gap between the two slabs in order to tune the guided resonance wavelength. An optical set-up is used to trace the photonic crystals transmission and reflection spectrum, in order to know the guided resonance position. A supercontinuum source illuminates the device at a normal incidence angle for wavelength between 400 nm and 800 nm. High-resolution spectra are obtained with a CCD camera spectrometer. Different types of one-slab photonic crystals are analyzed with this approach: we observe guided resonance peaks near 550 nm, 615 nm and 700 nm. Finally, a quantum dots microdrop is placed on the photonic crystal. The quantum dots emission wavelength matches with the photonic crystal guided resonance. A hyperspectral fluorescence microscope excites quantum dots between 436 nm and 483 nm, detects emission greater than 500 nm and plots a fluorescence wavelength spectrum. This set-up measures and compares the fluorescence of the quantum dots placed on and next to the photonic crystals. Our results show that the fluorescence is 30 times higher on the photonic crystals, but the fluorescence wavelength corresponds neither to the quantum dots emission nor to the photonic crystal guided resonance. In conclusion, this master thesis project demonstrates that it is possible to fabricate two-slab photonic crystals in silicon nitride and to plot their transmission and reflection spectra in order to find their guided resonance position. A fluorescence enhancement is visible, but at a different wavelength than of the quantum dots.

[Spectral characteristics of dissolved organic matter released during the metabolic process of small medusa].

PubMed

Guo, Dong-Hui; Yi, Yue-Yuan; Zhao, Lei; Guo, Wei-Dong

2012-06-01

The metabolic processes of jellyfish can produce dissolved organic matter (DOM) which will influence the functioning of the aquatic ecosystems, yet the optical properties of DOM released by jellyfish are unknown. Here we report the absorption and fluorescence properties of DOM released by a medusa species Black fordia virginica during a 24 h incubation experiment. Compared with the control group, an obvious increase in the concentrations of dissolved organic carbon (DOC), absorption coefficient (a280) and total dissolved nitrogen (TDN) was observed in incubation group. This clearly demonstrated the release of DOM, chromophoric DOM (CDOM) and dissolved nutrients by B. virginica which feed on enough of Artemia sp. before the experiment. The increase in spectral slope ratio (SR) and decrease in humification index (HIX) indicated that the released DOM was less-humified and had relatively lower molecular weight. Parallel factor analysis (PARAFAC) decomposed the fluorescence matrices of DOM into three humic-like components (C1-C3) and one protein-like component (C4). The Fmax of two components (C2: < 250, 295/386 nm; C4: 275/334 nm) with the emission wavelength < 400 nm increased significantly during the metabolic process of B. virginica. However, the Fmax of the other two components with the emission wavelength > 400 nm showed little changes. Thus, we suggested a zooplankton index (ZIX) to trace and characterize the DOM excreted by metabolic activity of zooplankton, which is calculated as the ratio of the sum of Fmax of all fluorescence components with the emission wavelength < 400 nm to the sum of Fmax of the other components with the emission wavelength > 400 nm.

[Fluorescence spectra and quantum yield of TiO2 nanocrystals synthesized by alcohothermal method].

PubMed

Song, Cui-Hong; Li, Yan-Ting; Li, Jing; Wei, Yong-Ju; Hu, Yu-Zhu; Wei, Yu

2008-01-01

Fluorescence spectra and fluorescence quantum yield of TiO2 nanocrystals were studied. Using tetra n-butyl titanate as a starting material, a facile alcohothermal technique was used to synthesize TiO2 nanocrystals. As can be seen from the transmittance electron microscopy (TEM) image, TiO2 nanocrystals with a relatively uniform particle size distribution of < 10 nm are present in the transparent sol. The transparent sol presents a strong stable fluorescence emission with a maximum at 450 nm, which is greatly dependent on the size quantization effects, defect energy level and the surface state of TiO2 nanocrystals. The quantum yield (gamma) of TiO2 was determined by the relative comparison procedure, using freshly prepared analytical purity quinine sulfate in 0.05 mol x L(-1) H2SO4 as a relative quantum yield standard. The emission quantum yield of TiO2 nanocrystals prepared in alcoholic media was calculated to be about 0.20 at wavelengths ranging from 330 to 370 nm, which was much higher than the values reported in previous works. So, it is supposed that nano-TiO2 will be applied as a potential quantum dots fluorescence probe in biological analysis.

Fluorenone based fluorescent probe for selective "turn-on" detection of pyrophosphate and alanine

NASA Astrophysics Data System (ADS)

Daniel Thangadurai, T.; Nithya, I.; Manjubaashini, N.; Bhuvanesh, N.; Bharathi, G.; Nandhakumar, R.; Nataraj, D.

2018-06-01

To sense biologically important entities with different size and dimensions, a fluorenone based fluorescent receptor was designed and synthesized. Probe 1 displayed a distinct fluorescence enhancement emission at 565 nm for pyrophosphate and 530 nm for alanine in polar solvent. The fluorescence titration experiments confirm 1:1 stoichiometric ratio with high-binding constant and very low limit of detection (LoD) values. Receptor 1 showed a highly selective and sensitive recognition to HP2O73 - and to alanine over other competitive anions and amino acids. In addition, the fluorescence lifetime measurement and reversible binding study results support the practical importance of 1.

In vivo assessment of wound re-epithelialization by UV fluorescence excitation imaging

NASA Astrophysics Data System (ADS)

Wang, Ying; Ortega-Martinez, Antonio; Padilla-Martinez, Juan Pablo; Williams, Maura; Farinelli, William; Anderson, R. R.; Franco, Walfre

2017-02-01

Background and Objectives: We have previously demonstrated the efficacy of a non-invasive, non-contact, fast and simple but robust fluorescence imaging (u-FEI) method to monitor the healing of skin wounds in vitro. This system can image highly-proliferating cellular processes (295/340 nm excitation/emission wavelengths) to study epithelialization in a cultured wound model. The objective of the current work is to evaluate the suitability of u-FEI for monitoring wound re-epithelialization in vivo. Study Design: Full-thickness wounds were created in the tail of rats and imaged weekly using u-FEI at 295/340nm excitation/emission wavelengths. Histology was used to investigate the correlation between the spatial distribution and intensity of fluorescence and the extent of wound epithelialization. In addition, the expression of the nuclear protein Ki67 was used to confirm the association between the proliferation of keratinocyte cells and the intensity of fluorescence. Results: Keratinocytes forming neo-epidermis exhibited higher fluorescence intensity than the keratinocytes not involved in re-epithelialization. In full-thickness wounds the fluorescence first appeared at the wound edge where keratinocytes initiated the epithelialization process. Fluorescence intensity increased towards the center as the keratinocytes partially covered the wound. As the wound healed, fluorescence decreased at the edges and was present only at the center as the keratinocytes completely covered the wound at day 21. Histology demonstrated that changes in fluorescence intensity from the 295/340nm band corresponded to newly formed epidermis. Conclusions: u-FEI at 295/340nm allows visualization of proliferating keratinocyte cells during re-epithelialization of wounds in vivo, potentially providing a quantitative, objective and simple method for evaluating wound closure in the clinic.

[Fluorescence characterization of dissolved organic matter in the East China Sea after diatom red tide dispersion].

PubMed

Zhuo, Peng-ji; Zhao, Wei-hong

2009-05-01

Fluorescence excitation-emission spectroscopy (EEMS) was employed to analyze the 3-dimensional fluorescence of dissolved organic matter in the East China Sea after diatom red tide dispersion. The relationships between fluorescence peak intensity, and salinity and chlorophyll-a were discussed. The centers of protein-like fluorescence peaks dispersed at Exmax/Exmax = 270-280/290-315 nm (Peak B), 220-230/290-305 nm (Peak D), 230-240/335-350 nm (Peak S) and 280/320 nm (Peak T). Two humic-like peaks appeared at 255-270/435-480 nm (Peak A)and 330-350/420-480 nm (Peak C). High tyrosine-like intensity was observed in diatom red tide dispersion area, and tryptophan-like fluorescence was also found which was lower. High FIB/FIS showed that diatom red tide produced much tyrosine-like matter during dispersion. Peaks S, A and C had positive correlation with one another, and their distributions were similar, which decreased with distance increasing away from the shore. Good negative correlations between peaks S, A and C and salinity suggested that Jiangsu-Zhejiang coastal water was the same source of them. Correlations between fluorescence peak intensity and chlorophyll-a were not remarkable enough to clear the relationship between fluorescence and living algal matter. It was supposed that the living algal matter contributed little to the fluorescence intensity of algal dispersion seawater.

Rescue of Injured Myocytes

DTIC Science & Technology

1990-11-30

mobimane ( mBBr ) have been used previously to measure intracellular glutathione and pro- tein thiols by flow cytometry (J Biol Chem 263, 14107). Here...200 p.M mBCl or 500 AM mBBr . Fluorescence was im- aged at 380 nm excitation and 470 nm emission using MDVM. Intracelluiar distribution of...predominantly glutathione) was 92% localized in cytosol. mBBr fluorescence released by 10 AM digitonin corresponded principally to glutathione

Labeling the oily core of nanocapsules and lipid-core nanocapsules with a triglyceride conjugated to a fluorescent dye as a strategy to particle tracking in biological studies

NASA Astrophysics Data System (ADS)

Fiel, Luana Almeida; Contri, Renata Vidor; Bica, Juliane Freitas; Figueiró, Fabrício; Battastini, Ana Maria Oliveira; Guterres, Sílvia Stanisçuaski; Pohlmann, Adriana Raffin

2014-05-01

The synthesis of novel fluorescent materials represents a very important step to obtain labeled nanoformulations in order to evaluate their biological behavior. The strategy of conjugating a fluorescent dye with triacylglycerol allows that either particles differing regarding supramolecular structure, i.e., nanoemulsions, nanocapsules, lipid-core nanocapsules, or surface charge, i.e., cationic nanocapsules and anionic nanocapsules, can be tracked using the same labeled material. In this way, a rhodamine B-conjugated triglyceride was obtained to prepare fluorescent polymeric nanocapsules. Different formulations were obtained, nanocapsules (NC) or lipid-core nanocapsules (LNC), using the labeled oil and Eudragit RS100, Eudragit S100, or poly(caprolactone) (PCL), respectively. The rhodamine B was coupled with the ricinolein by activating the carboxylic function using a carbodiimide derivative. Thin layer chromatography, proton nuclear magnetic resonance (1H-NMR), Fourier transform infrared spectroscopy (FTIR), UV-vis, and fluorescence spectroscopy were used to identify the new product. Fluorescent nanocapsule aqueous suspensions were prepared by the solvent displacement method. Their pH values were 4.6 (NC-RS100), 3.5 (NC-S100), and 5.0 (LNC-PCL). The volume-weighted mean diameter ( D 4.3) and polydispersity values were 150 nm and 1.05 (NC-RS100), 350 nm and 2.28 (NC-S100), and 270 nm and 1.67 (LNC-PCL). The mean diameters determined by photon correlation spectroscopy (PCS) ( z-average) were around 200 nm. The zeta potential values were +5.85 mV (NC-RS100), -21.12 mV (NC-S100), and -19.25 mV (LNC-PCL). The wavelengths of maximum fluorescence emission were 567 nm (NC-RS100 and LNC-PCL) and 574 nm (NC-S100). Fluorescence microscopy was used to evaluate the cell uptake (human macrophage cell line) of the fluorescent nanocapsules in order to show the applicability of the approach. When the cells were treated with the fluorescent nanocapsules, red emission was detected around the cell nucleus. We demonstrated that the rhodamine B-conjugated triglyceride is a promising new material to obtain versatile dye-labeled nanocarriers presenting different chemical nature in their surfaces.

Highly Sensitive Ratiometric Fluorescent Sensor for Trinitrotoluene Based on the Inner Filter Effect between Gold Nanoparticles and Fluorescent Nanoparticles.

PubMed

Lu, Hongzhi; Quan, Shuai; Xu, Shoufang

2017-11-08

In this work, we developed a simple and sensitive ratiometric fluorescent assay for sensing trinitrotoluene (TNT) based on the inner filter effect (IFE) between gold nanoparticles (AuNPs) and ratiometric fluorescent nanoparticles (RFNs), which was designed by hybridizing green emissive carbon dots (CDs) and red emissive quantum dots (QDs) into a silica sphere as a fluorophore pair. AuNPs in their dispersion state can be a powerful absorber to quench CDs, while the aggregated AuNPs can quench QDs in the IFE-based fluorescent assays as a result of complementary overlap between the absorption spectrum of AuNPs and emission spectrum of RFNs. As a result of the fact that TNT can induce the aggregation of AuNPs, with the addition of TNT, the fluorescent of QDs can be quenched, while the fluorescent of CDs would be recovered. Then, ratiometric fluorescent detection of TNT is feasible. The present IFE-based ratiometric fluorescent sensor can detect TNT ranging from 0.1 to 270 nM, with a detection limit of 0.029 nM. In addition, the developed method was successfully applied to investigate TNT in water and soil samples with satisfactory recoveries ranging from 95 to 103%, with precision below 4.5%. The simple sensing approach proposed here could improve the sensitivity of colorimetric analysis by changing the ultraviolet analysis to ratiometric fluorescent analysis and promote the development of a dual-mode detection system.

A simple ratiometric fluorescent sensor for fructose based on complexation of 10-hydroxybenzo[h]quinoline with boronic acid

NASA Astrophysics Data System (ADS)

Li, Huihui; Yang, Cailing; Zhu, Xinyue; Zhang, Haixia

2017-06-01

A simple ratiometric fluorescent sensor for fructose was presented. It consisted of 10-hydroxybenzo[h]quinoline (HBQ) which showed emission at 572 nm and 3-pyridylboronic acid (PDBA) whose complex with HBQ gave emission at 500 nm. The reaction of fructose with PDBA inhibited the complexation of HBQ with PDBA, resulting in the change of dual-emission intensity ratio. The sensor well quantified fructose in the range of 0.015-2.5 mM with detection limit of 0.005 mM. Besides, this sensor exhibited excellent selectivity and was successfully applied to fructose detection in food. This work provides a simple ratiometric sensing platform for sensitive and selective detection of fructose.

Noncontact point spectroscopy guided by two-channel fluorescence imaging in a hamster cheek pouch model

NASA Astrophysics Data System (ADS)

Yang, Victor X.; Yeow, Jenny; Lilge, Lothar D.; Kost, James; Mang, Thomas S.; Wilson, Brian C.

1999-07-01

A system for in vivo, fluorescence image-guided, non-contact point fluorescence spectroscopy is presented. A 442 nm HeCd laser is used as the fluorescence excitation source. An intensified CCD serves as the detector for both imaging and spectroscopy, on which two regions of 300 X 300 pixels were used for green (500 +/- 18 nm) and red (630 +/- 18 nm) imaging channels, and a strip of 600 X 120 pixels are used for emission spectroscopy (450 - 750 nm). At a working distance of 40 mm, the system has a spatial resolution of 0.16 mm and a spectral resolution of 5 nm. System performance is demonstrated in a carcinogenesis model in hamsters, where tumors were induced by painting DMBA in the cheek pouch. Autofluorescence and Photofrin-induced fluorescence measurements were performed every 2 weeks during the 18 weeks of tumor induction. Punch biopsies on selected animals were taken for histological staging. The results show that autofluorescence fluorescence can distinguish dysplasia from normal mucosal tissue model, utilizing the peak red intensity (or the red-to-green intensity ratio). Photofrin-induced fluorescence was superior to autofluorescence for differentiating high grade dysplasia from invasive cancer.

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

NASA Astrophysics Data System (ADS)

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

2017-03-01

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

Detection of low-concentration ammonia using differential laser-induced fluorescence on vapochromic coordination polymers

NASA Astrophysics Data System (ADS)

Yin, Dawei; Chapman, Glenn H.; Stevens, David; Gray, Bonnie; Leznoff, Daniel

2018-02-01

The detection of ammonia in parts per millions range has been challenging in sensors research, and is of great importance for industrial applications. In previous literature, Vapochromic Coordination Polymers (VCP) were developed to achieve luminescence upon a targeted gas exposures. We investigate a specific VCP, Zn[Au(CN)2]2,as an ammonia sensing material. Upon high concentration ammonia exposure, the fluorescent peak under near-UV stimulation undergoes a spectral shift from 460nm to 520nm, while the intensity increases by 3 4X. However, at ammonia concentrations < 50ppm, the spectral shift becomes hidden within the overall changing fluorescent spectrum shape. Then simple methods, such as detecting the peak wavelength or subtracting post-exposure from pre-exposure spectrums do not work. We developed further excitation and data processing techniques to detect ammonia at lower concentrations. A low-cost 405nm blue-ray DVD laser diode was used as the excitation source, providing a narrow band-width (4nm) stimulation that is separated from the emission peak. We measured the emission using a portable spectrometer (Photon Control SPM-002), and processed the data by separating the spectrum into two regions; (A) from 425 nm to 460 nm and (B) from 460nm to 500nm. Next, the integrated emissions under both regions were computed, and the value of shorter wavelength region (A) was subtracted from the longer wavelength one (B). When exposed to ammonia, region (A) reduces overall intensity while region (B) increases, resulting a signal starting from negative value and gradually increases to positive values, enabling the detection of 5ppm ammonia in less than 30 seconds gas exposure.

An organic dye with very large Stokes-shift and broad tunability of fluorescence: Potential two-photon probe for bioimaging and ultra-sensitive solid-state gas sensor

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

He, Tingchao; Tian, Xiaoqing; Lin, Xiaodong, E-mail: linxd@szu.edu.cn, E-mail: hdsun@ntu.edu.sg

Light-emitting nonlinear optical molecules, especially those with large Stokes shifts and broad tunability of their emission wavelength, have attracted considerable attention for various applications including biomedical imaging and fluorescent sensors. However, most fluorescent chromophores have only limited potential for such applications due to small Stokes shifts, narrow tunability of fluorescence emissions, and small optical nonlinearity in highly polar solvents. In this work, we demonstrate that a two-photon absorbing stilbene chromophore exhibits a large two-photon absorption action cross-section (ηδ = 320 GM) in dimethylsulfoxide (DMSO) and shows broad fluorescence tunability (125 nm) by manipulating the polarity of the surrounding medium. Importantly, a very large Stokesmore » shift of up to 227 nm is achieved in DMSO. Thanks to these features, this chromophore can be utilized as a two-photon probe for bioimaging applications and in an ultrasensitive solid-state gas detector.« less

Characterization of fluorescent-dissolved organic matter and identification of specific fluorophores in textile effluents.

PubMed

Li, Wentao; Xu, Zixiao; Wu, Qian; Li, Yan; Shuang, Chendong; Li, Aimin

2015-03-01

This study focused on the characterization of fluorescent-dissolved organic matter and identification of specific fluorophores in textile effluents. Samples from different textile wastewater treatment plants were characterized by high-performance liquid chromatography and size exclusion chromatography as well as fluorescence excitation-emission matrix spectra. Despite the highly heterogeneous textile effluents, the fluorescent components and their physicochemical properties were found relatively invariable, which is beneficial for the combination of biological and physicochemical treatment processes. The humic-like substance with triple-excitation peaks (excitation (Ex) 250, 310, 365/emission (Em) 460 nm) presented as the specific fluorescence indicator in textile effluents. It was also the major contributor to UV absorbance at 254 nm and resulted in the brown color of biologically treated textile effluents. By spectral comparison, the specific fluorophore in textile effluents could be attributed to the intermediate structure of azo dyes 1-amino-2-naphthol, which was transferred into the special humic-like substances during biological treatment.

A microwave-assisted solution combustion synthesis to produce europium-doped calcium phosphate nanowhiskers for bioimaging applications.

PubMed

Wagner, Darcy E; Eisenmann, Kathryn M; Nestor-Kalinoski, Andrea L; Bhaduri, Sarit B

2013-09-01

Biocompatible nanoparticles possessing fluorescent properties offer attractive possibilities for multifunctional bioimaging and/or drug and gene delivery applications. Many of the limitations with current imaging systems center on the properties of the optical probes in relation to equipment technical capabilities. Here we introduce a novel high aspect ratio and highly crystalline europium-doped calcium phosphate nanowhisker produced using a simple microwave-assisted solution combustion synthesis method for use as a multifunctional bioimaging probe. X-ray diffraction confirmed the material phase as europium-doped hydroxyapatite. Fluorescence emission and excitation spectra and their corresponding peaks were identified using spectrofluorimetry and validated with fluorescence, confocal and multiphoton microscopy. The nanowhiskers were found to exhibit red and far red wavelength fluorescence under ultraviolet excitation with an optimal peak emission of 696 nm achieved with a 350 nm excitation. Relatively narrow emission bands were observed, which may permit their use in multicolor imaging applications. Confocal and multiphoton microscopy confirmed that the nanoparticles provide sufficient intensity to be utilized in imaging applications. Copyright © 2013 Acta Materialia Inc. Published by Elsevier Ltd. All rights reserved.

Photoluminescent properties of Eu-doped ZnLiNbO4

NASA Astrophysics Data System (ADS)

Huang, Meng-Hsi; Lin, Ming-Hong; Fang, Te-Hua; Chang, Chia-Wei

2018-04-01

In this study, fluorescent Eu3+-doped ZnLiNbO4 materials were prepared via a vibrating milled solid-state reaction method. The objective was to develop new fluorescent oxide materials and study their fluorescent properties. The ZnLiNbO4 tetragonal spinel structure was formed with a single phase at a sintering temperature of 800 °C and with a regular shape at 1000 °C. The main exciton band was at 466 nm (7F0 → 5D2), and the main emission band was at 615 nm (5D0 → 7F2), which was an orange–red light band. The emission intensity was approximately 5% when the doping concentration reached 7%. The decay time was 2.96 ms.

Origin of the F685 and F695 fluorescence in photosystem II.

PubMed

Andrizhiyevskaya, Elena G; Chojnicka, Agnieszka; Bautista, James A; Diner, Bruce A; van Grondelle, Rienk; Dekker, Jan P

2005-06-01

The emission spectra of CP47-RC and core complexes of Photosystem II (PS II) were measured at different temperatures and excitation wavelengths in order to establish the origin of the emission and the role of the core antenna in the energy transfer and charge separation processes in PS II. Both types of particles reveal strong dependences of spectral shape and yield on temperature. The results indicate that the well-known F-695 emission at 77 K arises from excitations that are trapped on a red-absorbing CP47 chlorophyll, whereas the F-685 nm emission at 77 K arises from excitations that are transferred slowly from 683 nm states in CP47 and CP43 to the RC, where they are trapped by charge separation. We conclude that F-695 at 77 K originates from the low-energy part of the inhomogeneous distribution of the 690 nm absorbing chlorophyll of CP47, while at 4 K the fluorescence originates from the complete distribution of the 690 nm chlorophyll of CP47 and from the low-energy part of the inhomogeneous distribution of one or more CP43 chlorophylls.

Cutaneous porphyrins exhibit anti-stokes fluorescence that is detectable in sebum (Conference Presentation)

NASA Astrophysics Data System (ADS)

Tian, Giselle; Zeng, Haishan; Zhao, Jianhua; Wu, Zhenguo; Al Jasser, Mohammed; Lui, Harvey; Mclean, David I.

2016-02-01

Porphyrins produced by Propionibacterium acnes represent the principal fluorophore associated with acne, and appear as orange-red luminescence under the Wood's lamp. Assessment of acne based on Wood's lamp (UV) or visible light illumination is limited by photon penetration depth and has limited sensitivity for earlier stage lesions. Inducing fluorescence with near infrared (NIR) excitation may provide an alternative way to assess porphyrin-related skin disorders. We discovered that under 785 nm CW laser excitation PpIX powder exhibits fluorescence emission in the shorter wavelength range of 600-715 nm with an intensity that is linearly dependent on the excitation power. We attribute this shorter wavelength emission to anti-Stokes fluorescence. Similar anti-Stokes fluorescence was also detected focally in all skin-derived samples containing porphyrins. Regular (Stokes) fluorescence was present under UV and visible light excitation on ex vivo nasal skin and sebum from uninflamed acne, but not on nose surface smears or sebum from inflamed acne. Co-registered CW laser-excited anti-Stokes fluorescence and fs laser-excited multi-photon fluorescence images of PpIX powder showed similar features. In the skin samples because of the anti-Stokes effect, the NIR-induced fluorescence was presumably specific for porphyrins since there appeared to be no anti-Stokes emission signals from other typical skin fluorophores such as lipids, keratins and collagen. Anti-Stokes fluorescence under NIR CW excitation is more sensitive and specific for porphyrin detection than UV- or visible light-excited regular fluorescence and fs laser-excited multi-photon fluorescence. This approach also has higher image contrast compared to NIR fs laser-based multi-photon fluorescence imaging. The anti-Stokes fluorescence of porphyrins within sebum could potentially be applied to detecting and targeting acne lesions for treatment via fluorescence image guidance.

Noninvasive monitoring of intraocular pharmacokinetics of daunorubicin using fluorophotometry.

PubMed

Kizhakkethara, I; Li, X; el-Sayed, S; Khoobehi, B; Moshfeghi, D M; Rahimy, M; Peyman, G A

Daunorubicin is a cytotoxic drug, which, in nontoxic doses, is effective in preventing cellular proliferation in experimental vitreoretinopathy. We studied dose and clearance of daunorubicin in various ocular tissues using fluorophotometry techniques. In vitro tests: The emission of fluorescence from the daunorubicin solution having a concentration range of 0.1 to 10 micrograms/mL in phosphate buffer was measured using an excitation wavelength range of 489 +/- 10 nm. The emission of fluorescence was measured at 514 nm; the linearity of the response was determined using linear regression analysis. There is a fluorescence peak of daunorubicin at 485 nm. The validity and reproducibility of the method were examined. In vivo tests: The rabbits were randomized into three groups and daunorubicin concentrations of 4, 6, or 8 micrograms/mL were injected into the vitreous. Fluorophotometry scanning from the retina to the anterior chamber was performed with a commercially available fluorophotometer at various times up to 48 hours after injection to quantify fluorescence emission of daunorubicin. The standard curve of fluorescence versus concentration of daunorubicin was linear in the range of 0.1 to 8 micrograms/mL. It was sensitive up to 0.1 microgram. The daunorubicin time concentration profile showed a dose response relationship over the 48-hour period studied. The half-life of daunorubicin in the vitreous was about 5 hours. We performed fluorophotometry using a fluorophotometer whose exciter emits light at 489 nm, which is very close to an absorption peak of daunorubicin. These two values are close enough to obviate the need for modifying the commercial fluorophotometer. Therefore the concentration of daunorubicin in the vitreous cavity can be measured noninvasively.

Net Production and Consumption of Fluorescent Colored Dissolved Organic Matter by Natural Bacterial Assemblages Growing on Marine Phytoplankton Exudates▿

PubMed Central

Romera-Castillo, Cristina; Sarmento, Hugo; Álvarez-Salgado, Xosé Antón; Gasol, Josep M.; Marrasé, Celia

2011-01-01

An understanding of the distribution of colored dissolved organic matter (CDOM) in the oceans and its role in the global carbon cycle requires a better knowledge of the colored materials produced and consumed by marine phytoplankton and bacteria. In this work, we examined the net uptake and release of CDOM by a natural bacterial community growing on DOM derived from four phytoplankton species cultured under axenic conditions. Fluorescent humic-like substances exuded by phytoplankton (excitation/emission [Ex/Em] wavelength, 310 nm/392 nm; Coble's peak M) were utilized by bacteria in different proportions depending on the phytoplankton species of origin. Furthermore, bacteria produced humic-like substances that fluoresce at an Ex/Em wavelength of 340 nm/440 nm (Coble's peak C). Differences were also observed in the Ex/Em wavelengths of the protein-like materials (Coble's peak T) produced by phytoplankton and bacteria. The induced fluorescent emission of CDOM produced by prokaryotes was an order of magnitude higher than that of CDOM produced by eukaryotes. We have also examined the final compositions of the bacterial communities growing on the exudates, which differed markedly depending on the phytoplankton species of origin. Alteromonas and Roseobacter were dominant during all the incubations on Chaetoceros sp. and Prorocentrum minimum exudates, respectively. Alteromonas was the dominant group growing on Skeletonema costatum exudates during the exponential growth phase, but it was replaced by Roseobacter afterwards. On Micromonas pusilla exudates, Roseobacter was replaced by Bacteroidetes after the exponential growth phase. Our work shows that fluorescence excitation-emission matrices of CDOM can be a helpful tool for the identification of microbial sources of DOM in the marine environment, but further studies are necessary to explore the association of particular bacterial groups with specific fluorophores. PMID:21742918

An experimental study of the electronic absorption and fluorescence spectral properties of new p-substituted-N-phenylpyrroles and their electrosynthesized polymers.

PubMed

Diaw, A K D; Gningue-Sall, D; Yassar, A; Brochon, J-C; Henry, E; Aaron, J-J

2015-01-25

Electronic absorption and fluorescence spectral properties of new p-substituted-N-phenylpyrroles (N-PhPys), including HOPhPy, MeOPhPy, ThPhPy, PhDPy, DPhDPy, PyPhThThPhPy, and their available, electrosynthesized polymers were investigated. Electronic absorption spectra, fluorescence excitation and emission spectra, fluorescence quantum yields (ΦF) and lifetimes (τF), and other photophysical parameters of these N-PhPy derivatives and their polymers were measured in DMF, DMSO diluted solutions and/or solid state at room temperature. The electronic absorption spectra of N-PhPy derivatives and their polymers included one to several bands, located in the 270-395 nm region, according to the p-phenyl substituent electron-donating effect and conjugated heteroaromatic system length. The fluorescence excitation spectra were characterized by one broad main peak, with, in most cases, one (or more) poorly resolved shoulder (s), appearing in the 270-405 nm region, and their emission spectra were generally constituted of several bands located in the 330-480 nm region. No significant shift of the absorption, fluorescence excitation and emission spectra wavelengths was found upon going from the monomers to the corresponding polymers. ΦF values were high, varying between 0.11 and 0.63, according to the nature of substituents(s) and to the conjugated system extension. Fluorescence decays were mono-exponential for the monomers and poly-exponential for PyPhThThPhPy and for polymers. τF values were relatively short (0.35-5.17 ns), and markedly decreased with the electron-donor character of the phenyl group p-substituent and the conjugated system extension. Copyright © 2014 Elsevier B.V. All rights reserved.

An efficient ratiometric fluorescence sensor based on metal-organic frameworks and quantum dots for highly selective detection of 6-mercaptopurine.

PubMed

Jin, Meng; Mou, Zhao-Li; Zhang, Rui-Ling; Liang, Si-Si; Zhang, Zhi-Qi

2017-05-15

The development of a simple and accurate quantitative method for the determination of 6-mercaptopurine (6-MP) is of great importance because of its serious side effects. Ratiometric fluorescence (RF) sensors are not subject to interference from environmental factors, and exhibit enhanced precision and accuracy. Therefore, a novel RF sensor for the selective detection of 6-MP was developed based on a dual-emission nanosensor. The nanosensor was fabricated by combining a blue-emission metal-organic framework (MOF) NH 2 -MIL-53(Al) (λ em =425nm) with green-emission 3-mercaptopropionic acid-capped CdTe quantum dots (MPA-CdTe QDs) (λ em =528nm) under a single excitation wavelength (335nm). Upon addition of 6-MP, the fluorescence of NH 2 -MIL-53(Al) in the nanohybrid was selectively quenched due to strong inner filter effects, while the fluorescence of the MPA-CdTe QDs was enhanced. The novel RF sensor exhibited higher selectivity towards 6-MP than CdTe QDs alone, and higher sensitivity than MOFs alone. 6-MP could be detected in the range of 0-50μM with a detection limit of 0.15μM (S/N=3). The developed sensor was applied for the determination of 6-MP in human urine samples and satisfactory results were obtained. Overall, a novel and efficient fluorescence-based method was developed for the detection of 6-MP in biosamples. Copyright © 2016 Elsevier B.V. All rights reserved.

Detection of Free Thiols and Fluorescence Response of Phycoerythrin Chromophore after Ultraviolet-B Radiation Stress.

PubMed

Kannaujiya, Vinod K; Sinha, Rajeshwar P

2017-03-01

The chemistry of thiol-chromophore linkage plays a central role in the nature of fluorescence of phycoerythrin (PE). Interaction of thiol and chromophore is crucial for the energy transfer, redox signal and inhibition of oxidative damage. In the present investigation the effects of ultraviolet-B radiation on an emission fluorescence intensity and wavelength shift in PE due to interaction between thiol and chromophore by remarkable strategy of detection technique was studied. Purification of PE was done by using a gel permeation and ion exchange chromatography that yielded a quite high purity index (6.40) in a monomeric (αβ) form. UV-B radiation accelerated the quenching efficiency (24.9 ± 1.52%) by reducing fluorescence emission intensity of thiol linked chromophore after 240 min of UV-B exposure. However, after blocking of transiently released free thiol by N-ethylmaleimide, quenching efficiency was increased (36.8 ± 2.80%) with marked emission wavelength shift towards shorter wavelengths up to 562 nm as compared to 575 nm in control. Emission fluorescence of free thiol was at maximum after 240 min that was detected specifically by monobromobimane (mBrB) molecular probe. The association/dissociation of bilin chromophore was analyzed by SDS- and Native-PAGE that also indicated a complete reduction in emission fluorescence. Our work clearly shows an early detection of free thiols and relative interaction with chromophore after UV-B radiation which might play a significant role in structural and functional integrity of terminal PE.

Time-resolved fluorescence spectroscopy of human brain tumors

NASA Astrophysics Data System (ADS)

Marcu, Laura; Thompson, Reid C.; Garde, Smita; Sedrak, Mark; Black, Keith L.; Yong, William H.

2002-05-01

Fluorescence spectroscopy of the endogenous emission of brain tumors has been researched as a potentially important method for the intraoperative localization of brain tumor margins. In this study, we investigate the use of time-resolved laser-induced fluorescence spectroscopy (TR-LIFS) for demarcation of primary brain tumors by studying the time-resolved spectra of gliomas of different histologic grades. Time-resolved fluorescence (3 ns, 337 nm excitation) from excised human brain tumor show differences between the time-resolved emission of malignant glioma and normal brain tissue (gray and white matter). Our findings suggest that brain tumors can be differentiated from normal brain tissue based upon unique time-resolved fluorescence signature.

Development of transition metal dichalcogenide based quantum dots for light emitting diodes

NASA Astrophysics Data System (ADS)

Seth, Subhashree; Sharma, S. K.

2018-05-01

Photoluminescent quantum dots (QDs) were synthesized by facile colloidal chemical route. Its properties were characterized and analysed by utilizing Fluorescence, FTIR and UV-Vis spectrophotometers. The resultant MoS2 QD exhibits fluorescence at 470 nm for excitation wavelength 400 nm. The as prepared sample exhibits excitation dependent emission due to polydispersion of MoS2 in the dispersive medium which is the characteristics of colloidal synthesis. It is also observed that resultant MoS2 QDs show size tunable emission in the visible region. The FTIR spectrum confirms the attachment of oleic acid on the surface of MoS2. Absorption spectrum shows a band at 346 nm and a shoulder band at 400 nm. The band gap of quantum dots was obtained as 3.5 eV. CIE diagram indicates the shifting of colour coordinates towards green region with increasing excitation wavelength.

Fluorescence spectroscopy of soil pellets : The use of CP/PARAFAC.

NASA Astrophysics Data System (ADS)

Mounier, Stéphane; Nicolodeli, Gustavo; Redon, Roland; Hacherouf, Kalhed; Milori, Debora M. B. P.

2014-05-01

Fluorescence spectroscopy is one of the most sensitive techniques available for analytical purposes. It is relatively easy to implement, phenomenologically straightforward and well investigated. Largely non-invasive and fast, so that it can be useful for environmental applications. Fluorescence phenomenon is highly probable in molecular systems containing atoms with lone pairs of electrons such as C=O, aromatic, phenolic, quinone and more rigid unsaturated conjugated systems. These functional groups are present in humic substances (HS) from soils (Senesi, 1990; N. Senesi et al., 1991) and represent the main fluorophors of Soil Organic Matter (SOM). The extension of the conjugated electronic system, the level of heteroatom substitution and type and number of substituting groups under the aromatic rings strongly affect the intensity and wavelength of molecular fluorescence. However, to analyse the SOM it is generally done a chemical extraction that allows measuring the fluorescence response of the liquid extract. To avoid this fractionation of the SOM, Milori et al. (2006) proposed the application of laser induced fluorescence spectroscopy (LIFS) in whole soil. This work intends to assess the technical feasibility of 3D fluorescence spectroscopy using lamp for excitation to analyse solids opaque samples prepared with different substances. Seventy four (74) solid samples were prepared from different mixtures of boric acid (BA), humic substance acid and tryptophan (TRP) powder. The compounds were mixture and a pellet was done by using pressure (8 ton). The pellets were measured using a spectrofluorimeter HITACHI F4500, and a 3D fluorescence tensor was done from emission spectra (200-600 nm) with excitation range from 200 to 500 nm. The acquisition parameters were: step at 5 nm, scan speed at 2400 nm.min-1, response time at 0.1 s, excitation and emission slits at 5 nm and photomultiplier voltage at 700 V. Furthermore, measures of Laser-induced Fluorescence were performed in pellets (boric and humic acids mixture) using a portable system built by Embrapa Instrumentation. It comprises a diode laser (Coherent - CUBE) emitting at 405 nm (50 mW), and the detection of emission by a high sensitivity mini-spectrometer (USB4000 - Ocean Optics) using a range from 440 to 800 nm. In first step, the 3D tensors were then treated by the CP/PARAFAC algorithm to decompose the signal response after removing the diffusion signal : three components were extracted with a CORCONDIA over 60%. The first component can be associate an artefact of the measurement or boric acid fluorescence, the second and third component could the related to the two different fluorescence contributions of tryptophan molecule, one with central excitation/emission in 290/360 nm and other in 350/465 nm. The presence of a small quantity (i.e. few percent in mass) of humic acid (HA) is quenching drastically the TRP fluorescence. Complementary, measurements will be performed to understand this behaviour taking in account the absorption wavelength by the surface (colour) and by measuring the time life fluorescence of the samples. Humic acid fluorescence in pellets (BA and HA) cannot be observed using lamp + monochromator excitation due to low intensity of source. The same pellets were measure using LIFS system, and fluorescence intensity increased as a function of concentration of HA until occur the inner filter effect from 300 ppm, similar to the behaviour of HA in solution. Even whether solid surface measurements are easier, understanding is not yet clear. More investigation needs to be done. Moreover, it should be important to know if the use of CP/PARAFAC decomposition for such data is relevant with the trilinear model. References Milori, D.M.B.P., Galeti, H.V.A., Martin-Neto, L., Dieckow, J., González-Pérez, M., Bayer, C., Salton, J., 2006. Organic Matter Study of Whole Soil Samples Using Laser-Induced Fluorescence Spectroscopy. Soil Science Society of America Journal 70, 57. N. Senesi, TM, M., MR, P., Brunetti., G., 1991. Characterization, differentiation and classification of humic substances by fluorescence spectroscopy. Soil Science 152, 259-271. Senesi, N., 1990. Molecular and quantitative aspects of the chemistry of fulvic acid and its interactions with metal ions and organic chemicals. Analytica Chimica Acta 232, 77-106.

Measurement of fluorescence in a rhodamine-123 doped self-assembled "giant" mesostructured silica sphere using a smartphone as optical hardware.

PubMed

Canning, John; Lau, Angelica; Naqshbandi, Masood; Petermann, Ingemar; Crossley, Maxwell J

2011-01-01

The blue OLED emission from a mobile phone was characterised, revealing a sharp emission band centred at λ = 445 nm with a 3dB bandwidth Δλ ∼ 20 nm. It was used to excite Rhodamine 123 doped within a "giant" mesostructured silica sphere during fabrication through evaporative self-assembly of silica nanoparticles. Fluorescence was able to be detected using a standard optical microscope fitted with a green transmission pass filter and cooled CCD and with 1 ms exposure time demonstrating the potential of mobile platforms as the basis for portable diagnostics in the field.

Optofluidic chlorophyll lasers.

PubMed

Chen, Yu-Cheng; Chen, Qiushu; Fan, Xudong

2016-06-21

Chlorophylls are essential for photosynthesis and also one of the most abundant pigments on earth. Using an optofluidic ring resonator of extremely high Q-factors (>10(7)), we investigated the unique characteristics and underlying mechanism of chlorophyll lasers. Chlorophyll lasers with dual lasing bands at 680 nm and 730 nm were observed for the first time in isolated chlorophyll a (Chla). Particularly, a laser at the 730 nm band was realized in 0.1 mM Chla with a lasing threshold of only 8 μJ mm(-2). Additionally, we observed lasing competition between the two lasing bands. The presence of laser emission at the 680 nm band can lead to quenching or significant reduction of laser emission at the 730 nm band, effectively increasing the lasing threshold for the 730 nm band. Further concentration-dependent studies, along with theoretical analysis, elucidated the mechanism that determines when and why the laser emission band appears at one of the two bands, or concomitantly at both bands. Finally, Chla was exploited as the donor in fluorescence resonance energy transfer to extend the laser emission to the near infrared regime with an unprecedented wavelength shift as large as 380 nm. Our work will open a door to the development of novel biocompatible and biodegradable chlorophyll-based lasers for various applications such as miniaturized tunable coherent light sources and in vitro/in vivo biosensing. It will also provide important insight into the chlorophyll fluorescence and photosynthesis processes inside plants.

One-pot synthesis of biocompatible Te@phenol formaldehyde resin core-shell nanowires with uniform size and unique fluorescent properties by a synergized soft-hard template process.

PubMed

Qian, Haisheng; Zhu, Enbo; Zheng, Shunji; Li, Zhengquan; Hu, Yong; Guo, Changfa; Yang, Xingyun; Li, Liangchao; Tong, Guoxiu; Guo, Huichen

2010-12-10

One-pot hydrothermal process has been developed to synthesize uniform Te@phenol formaldehyde resin core-shell nanowires with unique fluorescent properties. A synergistic soft-hard template mechanism has been proposed to explain the formation of the core-shell nanowires. The Te@phenol formaldehyde resin core-shell nanowires display unique fluorescent properties, which give strong luminescent emission in the blue-violet and green regions with excitation wavelengths of 270 nm and 402 nm, respectively.

Selective and Sensitive Detection of Heavy Metal Ions in 100% Aqueous Solution and Cells with a Fluorescence Chemosensor Based on Peptide Using Aggregation-Induced Emission.

PubMed

Neupane, Lok Nath; Oh, Eun-Taex; Park, Heon Joo; Lee, Keun-Hyeung

2016-03-15

A fluorescent peptidyl chemosensor for the detection of heavy metal ions in aqueous solution as well as in cells was synthesized on the basis of the peptide receptor for the metal ions using an aggregation-induced emission fluorophore. The peptidyl chemosensor (1) bearing tetraphenylethylene fluorophore showed an exclusively selective turn-on response to Hg(2+) among 16 metal ions in aqueous buffered solution containing NaCl. The peptidyl chemosensor complexed Hg(2+) ions and then aggregated in aqueous buffered solution, resulting in the significant enhancement (OFF-On) of emissions at around 470 nm. The fluorescent sensor showed a highly sensitive response to Hg(2+), and about 1.0 equiv of Hg(2+) was enough for the saturation of the emission intensity change. The detection limit (5.3 nM, R(2) = 0.99) of 1 for Hg(2+) ions was lower than the maximum allowable level of Hg(2+) in drinking water by EPA. Moreover, the peptidyl chemosensor penetrated live cells and detected intracellular Hg(2+) ions by the turn-on response.

Spectral fluorescent properties of tissues in vivo with excitation in the red wavelength range

NASA Astrophysics Data System (ADS)

Stratonnikov, Alexander A.; Loschenov, Victor B.; Klimov, D. V.; Edinac, N. E.; Wolnukhin, V. A.; Strashkevich, I. A.

1997-12-01

The spectral fluorescence analysis is a promising method for differential tissue diagnostic. Usually the UV and visible light is used for fluorescence excitation with emission registration in the visible wavelength range. The light penetration length in this wavelength range is very small allowing one to analyze only the surface region of the tissue. Here we present the tissue fluorescent spectra in vivo excited in the red wavelength region. As excitation light source we used compact He-Ne laser (632.8 nm) and observed the fluorescence in 650 - 800 nm spectral range. The various tissues including normal skin, psoriasis, tumors, necrosis as well as photosensitized tissues have been measured.

Bench-Top Antigen Detection Technique that Utilizes Nanofiltration and Fluorescent Dyes which Emit and Absorb Light in the Near Infrared

NASA Technical Reports Server (NTRS)

Varaljay-Spence, Vanessa A.; Scardelletti, Maximilian C.

2007-01-01

This article discusses the development of a bench-top technique to detect antigens in fluids. The technique involves the use of near infrared NIR fluorescent dyes conjugated to antibodies, centrifugation, nanofilters, and spectrometry. The system used to detect the antigens utilizes a spectrometer, fiber optic cables, NIR laser, and laptop computer thus making it portable and ideally suited for desk top analysis. Using IgM as an antigen and the secondary antibody, anti-IgM conjugated to the near infrared dye, IRDye (trademark) 800, for detection, we show that nanofiltration can efficiently and specifically separate antibody-antigen complexes in solution and that the complexes can be detected by a spectrometer and software using NIR laser excitation at 778 nm and NIR dye offset emission at 804 nm. The peak power detected at 778 nm for the excitation emission and at 804 nm for the offset emission is 879 pW (-60.06 dBm) and 35.7 pW (-74.5 dBm), respectively.

Spectral Properties of Er3+/Tm3+ Co-Doped ZBLAN Glasses and Fibers

PubMed Central

Liao, Xili; Jiang, Xiaobo; Yang, Qiuhong; Wang, Longfei; Chen, Danping

2017-01-01

A series of Er3+/Tm3+ co-doped fluoride (ZBLAN) glasses and fibers was prepared and their fluorescence spectra was measured under excitation at 793 nm and 980 nm. Correlation between the self-absorption effect of rare-earth ions and the shift of the emission peak was investigated. With the increasing length of fiber, the emission peaks red-shift when self-absorption occurs at the upper level of emission transition or blue-shift when that occurs at the lower level. As a result of the strong self-absorption effect, Er3+/Tm3+ co-doped fibers mainly yield 1390–1470, 1850–1980, and 2625–2750 nm emissions when excited at 793 nm, and 1480–1580, 1800–1980, and 2625–2750 nm emissions when excited at 980 nm. Further, a broadband emission in the range of 1410–1580 nm covering the S + C communication band was obtained by the dual-pumping scheme of 793 nm and 980 nm. Results suggest that the dual-pumping scheme would be more effective and important for an Er3+/Tm3+ co-doped fiber amplifier working in the S + C communication band. PMID:28772846

Spectral Properties of Er3+/Tm3+ Co-Doped ZBLAN Glasses and Fibers.

PubMed

Liao, Xili; Jiang, Xiaobo; Yang, Qiuhong; Wang, Longfei; Chen, Danping

2017-05-03

A series of Er 3+ /Tm 3+ co-doped fluoride (ZBLAN) glasses and fibers was prepared and their fluorescence spectra was measured under excitation at 793 nm and 980 nm. Correlation between the self-absorption effect of rare-earth ions and the shift of the emission peak was investigated. With the increasing length of fiber, the emission peaks red-shift when self-absorption occurs at the upper level of emission transition or blue-shift when that occurs at the lower level. As a result of the strong self-absorption effect, Er 3+ /Tm 3+ co-doped fibers mainly yield 1390-1470, 1850-1980, and 2625-2750 nm emissions when excited at 793 nm, and 1480-1580, 1800-1980, and 2625-2750 nm emissions when excited at 980 nm. Further, a broadband emission in the range of 1410-1580 nm covering the S + C communication band was obtained by the dual-pumping scheme of 793 nm and 980 nm. Results suggest that the dual-pumping scheme would be more effective and important for an Er 3+ /Tm 3+ co-doped fiber amplifier working in the S + C communication band.

Development of infrared sensors using energy transfer/energy upconversion processes: Study of laser excited fluorescence in rare Earth ion doped crystals

NASA Technical Reports Server (NTRS)

Nash-Stevenson, S. K.; Reddy, B. R.; Venkateswarlu, P.

1994-01-01

A summary is presented of the spectroscopic study of three systems: LaF3:Ho(3+), LaF3:Er(3+) and CaF2:Nd(3+). When the D levels of Ho(3+) in LaF3 were resonantly excited with a laser beam of 640 nm, upconverted emissions were detected from J (416 nm), F (485 nm), and E (546 nm) levels. Energy upconverted emissions were also observed from F and E levels of Ho(3+) when the material was excited with an 800 nm near infrared laser. When the D levels of Er(3+) in LaF3 were resonantly excited with a laser beam of 637 nm, upconverted emissions were detected from the E (540 nm) and P (320, 400, and 468 nm) levels. Energy upconverted emissions were also observed from F, E, and D levels of Er(3+) when the material was resonantly excited with an 804 nm near infrared laser. When the D levels of Nd(3+) in CaF2 were resonantly excited with a laser beam of 577 nm, upconverted emissions were detected from the L (360 and 382 nm), K (418 nm), and I (432 nm) levels. Very weak upconverted emissions were detected when this system was irradiated with a near infrared laser. The numbers in parentheses are the wavelengths of the emissions.

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

NASA Astrophysics Data System (ADS)

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

2018-04-01

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

Mechanochemical synthesis of fluorescent carbon dots from cellulose powders

NASA Astrophysics Data System (ADS)

Chae, Ari; Ram Choi, Bo; Choi, Yujin; Jo, Seongho; Kang, Eun Bi; Lee, Hyukjin; Park, Sung Young; In, Insik

2018-04-01

A novel mechanochemical method was firstly developed to synthesize carbon nanodots (CNDs) or carbon nano-onions (CNOs) through high-pressure homogenization of cellulose powders as naturally abundant resource depending on the treatment times. While CNDs (less than 5 nm in size) showed spherical and amorphous morphology, CNOs (10-50 nm in size) presented polyhedral shape, and onion-like outer lattice structure, graphene-like interlattice spacing of 0.36 nm. CNOs showed blue emissions, moderate dispersibility in aqueous media, and high cell viability, which enables efficient fluorescence imaging of cellular media.

SU-G-IeP4-08: Initial Investigations of Up-Converting Nanoparticles (UCNP) for 3D Tissue Imaging in Optical-ECT

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

Yoon, S; Dewhirst, M; Oldham, M

Purpose: Near-IR absorptive up-converting nanoparticles (UCNPs) is a novel contrast for optical-ECT that allows auto-fluorescence-free 3D imaging of labeled cells in a matrix of large (∼1cm{sup 3}) unsectioned normal tissue. This has the potential to image small metastases or dormant cells that is difficult with down-converting fluorescing dyes due to auto-fluorescence. The feasibility of imaging UCNP in agarose phantoms and a mouse lung is demonstrated, aided by a 3D-printed optical-ECT stage designed to excite UCNP in a mouse lung. Methods: The UCNP, NaYF{sub 4}:Yb/Er (20/2%), studied in this work up-converts 980nm light to visible light peaking sharply at ∼540nm. Tomore » characterize the UCNP emission as a function of UCNP concentration, cylindrical 2.5%wt agarose phantoms infused with UCNP at concentrations of 25µg/mL and 50µg/mL were exposed to 1.5W 980nm laser coupled to an optical fiber. The fiber was held stably at 1cm above the stage via a custom 3D-printed stage. An optically cleared lung harvested from a BALBc mice was then injected with 100µL of 1mg/mL UCNP solution ex vivo. Tomographic imaging of the UCNP emission in lung was performed. Results: The laser beam tract is visualized within the agarose phantom. A line profile of UCNP emission at 25µg/mL versus 50µg/mL shows that increasing the UCNP concentration increases emission count. UCNPs injected into a cleared mouse lung disperse throughout the respiratory tract, allowing for visualization and 3D reconstruction. Excitation before and after UCNP injection shows the tissue exhibits no auto-fluorescence at 980nm, allowing clear view of the UCNP without any obscuring features such as conventional down-converting fluorescent tags. Conclusion: We confirm that up-conversion in tissue circumvents completely tissue auto-fluorescence, which allowed background-free 3D reconstruction of the UCNP distribution. We also confirm that raising the UCNP concentration increases emission and that UCNPs are retained in agarose samples during the optical clearing process.« less

Enhanced Fluorescence Emission of Me-ADOTA+ by Self-Assembled Silver Nanoparticles on a Gold Film

PubMed Central

Sørensen, Thomas J.; Laursen, Bo W.; Luchowski, Rafal; Shtoyko, Tanya; Akopova, Irina; Gryczynski, Zygmunt; Gryczynski, Ignacy

2009-01-01

We report a multi-fold enhancement of the fluorescence of methyl-azadioxatriangulenium chloride (Me-ADOTA•Cl) in PVA deposited on a 50 nm thick gold mirror carrying an evaporation induced self-assembly of colloidal silver nanoparticles (Ag-SACs). The average measured increase in fluorescence emission of about 50-fold is accompanied by hot spots with a local enhancement in brigthness close to 200. The long lifetime of the dye allows for the first direct determination of the correlation between the enhancement of emission intensity and the decrease in fluorescence lifetime. The Ag-SACs surface preparation and observed enhancements are highly reproducible. We believe that these robust plasmonic surfaces will find use in sensing platforms for ultrasensitive detection. PMID:20161182

Evolution of near UV Halley's spectrum in the inner coma

NASA Technical Reports Server (NTRS)

Rousselot, Phillippe; Clairemidi, Jacques; Vernotte, F.; Moreels, Guy

1992-01-01

A direct way to observe the photodissociation of water vapor in a cometary coma is to detect the OH prompt emission. This emission is shifted of delta lambda = 4 nm with respect to the OH 309 nm fluorescence band. The extended data set obtained with the three-channel spectrometer on-board Vega 2 reveals at short distance of the nucleus (i.e., less than 600 km) an excess of emission on the right wing of the OH band which may be interpreted as being mainly due to prompt emission.

A novel reaction-based colorimetric and ratiometric fluorescent sensor for cyanide anion with a large emission shift and high selectivity.

PubMed

Wang, Shaodan; Fei, Xiaoliang; Guo, Jing; Yang, Qingbiao; Li, Yaoxian; Song, Yan

2016-01-01

A hybrid carbazole-hemicyanine dye (Cac) has been developed as a novel colorimetric and ratiometric fluorescent sensor for cyanide detection. Upon treatment with cyanide, Cac displayed a remarkable fluorescence ratiometric response, with the emission wavelength displaying a very large emission shift (214 nm). The detection of cyanide was performed via the nucleophilic addition of cyanide anion to the indolium group of the sensor, which resulted in the blocking of the intramolecular charge transfer (ICT) process in the sensor, inducing a ratiometric fluorescence change and simultaneously an obvious color change. Furthermore, competitive anions did not showed any significant changes both in color and emission intensity ratio (I381/I595), indicating the high selectivity of the sensor to CN(-). Copyright © 2015 Elsevier B.V. All rights reserved.

Shortwave infrared fluorescence imaging with the clinically approved near-infrared dye indocyanine green.

PubMed

Carr, Jessica A; Franke, Daniel; Caram, Justin R; Perkinson, Collin F; Saif, Mari; Askoxylakis, Vasileios; Datta, Meenal; Fukumura, Dai; Jain, Rakesh K; Bawendi, Moungi G; Bruns, Oliver T

2018-04-24

Fluorescence imaging is a method of real-time molecular tracking in vivo that has enabled many clinical technologies. Imaging in the shortwave IR (SWIR; 1,000-2,000 nm) promises higher contrast, sensitivity, and penetration depths compared with conventional visible and near-IR (NIR) fluorescence imaging. However, adoption of SWIR imaging in clinical settings has been limited, partially due to the absence of US Food and Drug Administration (FDA)-approved fluorophores with peak emission in the SWIR. Here, we show that commercially available NIR dyes, including the FDA-approved contrast agent indocyanine green (ICG), exhibit optical properties suitable for in vivo SWIR fluorescence imaging. Even though their emission spectra peak in the NIR, these dyes outperform commercial SWIR fluorophores and can be imaged in the SWIR, even beyond 1,500 nm. We show real-time fluorescence imaging using ICG at clinically relevant doses, including intravital microscopy, noninvasive imaging in blood and lymph vessels, and imaging of hepatobiliary clearance, and show increased contrast compared with NIR fluorescence imaging. Furthermore, we show tumor-targeted SWIR imaging with IRDye 800CW-labeled trastuzumab, an NIR dye being tested in multiple clinical trials. Our findings suggest that high-contrast SWIR fluorescence imaging can be implemented alongside existing imaging modalities by switching the detection of conventional NIR fluorescence systems from silicon-based NIR cameras to emerging indium gallium arsenide-based SWIR cameras. Using ICG in particular opens the possibility of translating SWIR fluorescence imaging to human clinical applications. Indeed, our findings suggest that emerging SWIR-fluorescent in vivo contrast agents should be benchmarked against the SWIR emission of ICG in blood.

The endogenous fluorescence of fibroblast in collagen gels as indicator of stiffness of the extracellular matrix

NASA Astrophysics Data System (ADS)

Padilla-Martinez, J. P.; Ortega-Martinez, A.; Franco, W.

2016-03-01

The stiffness or rigidity of the extracellular matrix (ECM) regulates cell response. Established mechanical tests to measure stiffness, such as indentation and tensile tests, are invasive and destructive to the sample. Endogenous or native molecules to cells and ECM components, like tryptophan and cross-links of collagen, display fluorescence upon irradiation with ultraviolet light. Most likely, the concentration of these endogenous fluorophores changes as the stiffness of the ECM changes. In this work we investigate the endogenous fluorescence of collagen gels containing fibroblasts as a non-invasive non-destructive method to measure stiffness of the ECM. Human fibroblast cells were cultured in three-dimensional gels of type I collagen (50,000 cells/ml). This construct is a simple model of tissue contraction. During contraction, changes in the excitation-emission matrix (a fluorescence map in the 240-520/290-530 nm range) of constructs were measured with a spectrofluoremeter, and changes in stiffness were measured with a standard indentation test over 16 days. Results show that a progressive increase in fluorescence of the 290/340 nm excitation-emission pair correlates with a progressive increase in stiffness (r=0.9, α=0.5). The fluorescence of this excitation-emission pair is ascribed to tryptophan and variations in the fluorescence of this pair correlate with cellular proliferation. In this tissue model, the endogenous functional fluorescence of proliferating fibroblast cells is a biomechanical marker of stiffness of the ECM.

Changes in fluorescence spectra of bioaerosols exposed to ozone in a laboratory reaction chamber to simulate atmospheric aging.

PubMed

Santarpia, Joshua L; Pan, Yong-Le; Hill, Steven C; Baker, Neal; Cottrell, Brian; McKee, Laura; Ratnesar-Shumate, Shanna; Pinnick, Ronald G

2012-12-31

A laboratory system for exposing aerosol particles to ozone and rapidly measuring the subsequent changes in their single-particle fluorescence is reported. The system consists of a rotating drum chamber and a single-particle fluorescence spectrometer (SPFS) utilizing excitation at 263 nm. Measurements made with this system show preliminary results on the ultra-violet laser-induced-fluorescence (UV-LIF) spectra of single aerosolized particles of Yersinia rohdei, and of MS2 (bacteriophage) exposed to ozone. When bioparticles are exposed in the chamber the fluorescence emission peak around 330 nm: i) decreases in intensity relative to that of the 400-550 nm band; and ii) shifts slightly toward shorter-wavelengths (consistent with further drying of the particles). In these experiments, changes were observed at exposures below the US Environmental Protection Agency (EPA) limits for ozone.

Fluorescence fingerprint as an instrumental assessment of the sensory quality of tomato juices.

PubMed

Trivittayasil, Vipavee; Tsuta, Mizuki; Imamura, Yoshinori; Sato, Tsuneo; Otagiri, Yuji; Obata, Akio; Otomo, Hiroe; Kokawa, Mito; Sugiyama, Junichi; Fujita, Kaori; Yoshimura, Masatoshi

2016-03-15

Sensory analysis is an important standard for evaluating food products. However, as trained panelists and time are required for the process, the potential of using fluorescence fingerprint as a rapid instrumental method to approximate sensory characteristics was explored in this study. Thirty-five out of 44 descriptive sensory attributes were found to show a significant difference between samples (analysis of variance test). Principal component analysis revealed that principal component 1 could capture 73.84 and 75.28% variance for aroma category and combined flavor and taste category respectively. Fluorescence fingerprints of tomato juices consisted of two visible peaks at excitation/emission wavelengths of 290/350 and 315/425 nm and a long narrow emission peak at 680 nm. The 680 nm peak was only clearly observed in juices obtained from tomatoes cultivated to be eaten raw. The ability to predict overall sensory profiles was investigated by using principal component 1 as a regression target. Fluorescence fingerprint could predict principal component 1 of both aroma and combined flavor and taste with a coefficient of determination above 0.8. The results obtained in this study indicate the potential of using fluorescence fingerprint as an instrumental method for assessing sensory characteristics of tomato juices. © 2015 Society of Chemical Industry.

Fundus autofluorescence and the bisretinoids of retina.

PubMed

Sparrow, Janet R; Wu, Yalin; Nagasaki, Takayuki; Yoon, Kee Dong; Yamamoto, Kazunori; Zhou, Jilin

2010-11-01

Imaging of the human fundus of the eye with excitation wavelengths in the visible spectrum reveals a natural autofluorescence, that in a healthy retina originates primarily from the bisretinoids that constitute the lipofuscin of retinal pigment epithelial (RPE) cells. Since the intensity and distribution of fundus autofluorescence is altered in the presence of retinal disease, we have examined the fluorescence properties of the retinal bisretinoids with a view to aiding clinical interpretations. As is also observed for fundus autofluorescence, fluorescence emission from RPE lipofuscin was generated with a wide range of exciting wavelengths; with increasing excitation wavelength, the emission maximum shifted towards longer wavelengths and spectral width was decreased. These features are consistent with fluorescence generation from a mixture of compounds. While the bisretinoids that constitute RPE lipofuscin all fluoresced with maxima that were centered around 600 nm, fluorescence intensities varied when excited at 488 nm, the excitation wavelength utilized for fundus autofuorescence imaging. For instance the fluorescence efficiency of the bisretinoid A2-dihydropyridine-phosphatidylethanolamine (A2-DHP-PE) was greater than A2E and relative to both of the latter, all-trans-retinal dimer-phosphatidylethanolamine was weakly fluorescent. On the other hand, certain photooxidized forms of the bisretinoids present in both RPE and photoreceptor cells were more strongly fluorescent than the parent compound. We also sought to evaluate whether diffuse puncta of autofluorescence observed in some retinal disorders of monogenic origin are attributable to retinoid accumulation. However, two retinoids of the visual cycle, all-trans-retinyl ester and all-trans-retinal, did not exhibit fluorescence at 488 nm excitation.

Microwave-assisted one-step synthesis of white light-emitting carbon dot suspensions

NASA Astrophysics Data System (ADS)

Vanessa, Hinterberger; Wenshuo, Wang; Cornelia, Damm; Simon, Wawra; Martin, Thoma; Wolfgang, Peukert

2018-06-01

In this contribution, we demonstrate that an aqueous solution with adjustable fluorescent color, including white light emission, can be achieved by a rapid one-step microwave synthesis method resulting in a mixture of blue-emitting carbon dots (CDs) and the yellow-emitting 2,3-diaminophenazine (DAP). Aqueous mixtures of o-phenylene-diamine (oPD) and citric acid (CA) are used as precursors. The resulting product structures are analyzed by FT-IR and NMR spectroscopy and the size of the resulting CDs is determined by atomic force microscopy to be 1.1 ± 0.3 nm. The synthesized solution exhibits two fluorescence emission peaks at 430 and 560 nm, which were found to originate from the CDs and DAP, respectively. The intensity ratio of both fluorescence peaks depends on pH, which is driven by the protonation state of DAP. In consequence, the fluorescence emission color of the CD solution can be tuned precisely and reproducibly from blue to white to yellow by careful control of the pH. Finally, at a pH level of 5.4, at which there is equal blue and yellow emission intensity, a white light emitting solution can be successfully produced in a very fast and simple synthesis procedure.

Development of a wide-field fluorescence imaging system for evaluation of wound re-epithelialization

NASA Astrophysics Data System (ADS)

Franco, Walfre; Gutierrez-Herrera, Enoch; Purschke, Martin; Wang, Ying; Tam, Josh; Anderson, R. Rox; Doukas, Apostolos

2013-03-01

Normal skin barrier function depends on having a viable epidermis, an epithelial layer formed by keratinocytes. The transparent epidermis, which is less than a 100 mum thick, is nearly impossible to see. Thus, the clinical evaluation of re-epithelialization is difficult, which hinders selecting appropriate therapy for promoting wound healing. An imaging system was developed to evaluate epithelialization by detecting endogenous fluorescence emissions of cellular proliferation over a wide field of view. A custom-made 295 nm ultraviolet (UV) light source was used for excitation. Detection was done by integrating a near-UV camera with sensitivity down to 300 nm, a 12 mm quartz lens with iris and focus lock for the UV regime, and a fluorescence bandpass filter with 340 nm center wavelength. To demonstrate that changes in fluorescence are related to cellular processes, the epithelialization of a skin substitute was monitored in vitro. The skin substitute or construct was made by embedding microscopic live human skin tissue columns, 1 mm in diameter and spaced 1 mm apart, in acellular porcine dermis. Fluorescence emissions clearly delineate the extent of lateral surface migration of keratinocytes and the total surface covered by the new epithelium. The fluorescence image of new epidermis spatially correlates with the corresponding color image. A simple, user-friendly way of imaging the presence of skin epithelium would improve wound care in civilian burns, ulcers and surgeries.

Microwave-assisted synthesis of highly luminescent N- and S-co-doped carbon dots as a ratiometric fluorescent probe for levofloxacin.

PubMed

Li, Huiyu; Xu, Yuan; Ding, Jie; Zhao, Li; Zhou, Tianyu; Ding, Hong; Chen, Yanhua; Ding, Lan

2018-01-10

Uniform N- and S-co-doped carbon dots (NSCDs) with fluorescence quantum yields of up to 64% were synthesized via a one-step microwave-assisted method. Ammonium citrate and L-cysteine act as precursors, and synthesis is completed in 2.5 min using a 750 W microwave oven to give a 62% yield. The NSCDs show bright blue fluorescence (with excitation/emission peaks at 353/426 nm) and have narrow size distribution. On exposure to levofloxacin (LEV), the emission maximum shifts to 499 nm. This effect was used to design ratiometric (2-wavelength) assays for LEV. The fluorometric method (based on measurement of the fluorescence intensity ratio at 499 and 426 nm) has a detection limit of 5.1 μg·L -1 (3σ/k) and a linear range that extends from 0.01 to 70 mg·L -1 . The method was applied to the determination of LEV in three kinds of spiked water samples and has recoveries in the range from 98.6 to 106.8%. The fluorescent probe described here is highly selective and sensitive. Graphical Abstract Highly luminescent N- and S-co-doped carbon dots were synthesized using AC (ammonium citrate) and Cys (L-cysteine) by microwave-assisted method, and were applied to the visual and ratiometric fluorescence determination of LEV (levofloxacin).

Development of new devices for detection of gastric cancer on laparoscopic surgery using near-infrared light

NASA Astrophysics Data System (ADS)

Inada, Shunko A.; Fuchi, Shingo; Mori, Kensaku; Hasegawa, Junichi; Misawa, Kazunari; Nakanishi, Hayao

2015-03-01

In recent year, for the treatment of gastric cancer the laparoscopic surgery is performed, which has good benefits, such as low-burden, low-invasive and the efficacy is equivalent to the open surgery. For identify location of the tumor intraperitoneally for extirpation of the gastric cancer, several points of charcoal ink is injected around the primary tumor. However, in the time of laparoscopic operation, it is difficult to estimate specific site of primary tumor, because the injected charcoal ink diffusely spread to the area distant from the tumor in the stomach. Therefore, a broad area should be resected which results in a great stress for the patients. To overcome this problem, we focused in the near-infrared wavelength of 1000nm band which have high biological transmission. In this study, we developed a fluorescent clip which was realized with glass phosphor (Yb3+, Nd3+ doped to Bi2O3-B2O3 based glasses. λp: 976 nm, FWHM: 100 nm, size: 2x1x3 mm) and the laparoscopic fluorescent detection system for clip-derived near-infrared light. To evaluate clinical performance of a fluorescent clip and the laparoscopic fluorescent detection system, we used resected stomach (thickness: 13 mm) from the patients. Fluorescent clip was fixed on the gastric mucosa, and an excitation light (λ: 808 nm) was irradiated from outside of stomach for detection of fluorescence through stomach wall. As a result, fluorescence emission from the clip was successfully detected. Furthermore, we confirmed that detection sensitivity of the emission of fluorescence from the clip depends on the output power of the excitation light. We conformed that the fluorescent clip in combination with laparoscopic fluorescent detection system is very useful method to identify the exact location of the primary gastric cancer.

Fluorescence detection of protein content in house dust: the possible role of keratin.

PubMed

Voloshina, O V; Shirshin, E A; Lademann, J; Fadeev, V V; Darvin, M E

2017-03-01

We propose a fluorescence method for protein content assessment in fine house dust, which can be used as an indicator of the hygienic state of occupied rooms. The results of the measurements performed with 30 house dust samples, including ultrafiltration experiments, strongly suggest that the fluorescence emission of house dust extracts excited at 350 nm is mainly due to protein fragments, which are presumably keratin hydrolysates. This suggestion is supported by several facts: (i) Spectral band shapes for all the samples under investigation are close and correspond to that of keratin; (ii) fluorescence intensity correlates with the total protein content as provided by Lowry assay; (iii) treatment of the samples with proteinase K, which induces keratin hydrolysis, results in fluorescence enhancement without changing fluorescence band shape; and (iv) Raman spectra of keratin and fine house dust samples exhibit a very similar structure. Based on the obtained results and literature data, we propose a hypothesis that keratin is a major substrate for fluorescence species in fine house dust, which are responsible for emission at 350-nm excitation. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

Femtosecond laser fluorescence and propagation in very dense potassium vapor.

PubMed

Makdisi, Y; Kokaj, J; Afrousheh, K; Nair, R; Mathew, J; Pichler, G

2013-12-16

Femtosecond (fs) laser propagation and fluorescence of dense potassium vapor was studied, and the spectral region around the first and the second doublets of the principal series lines of potassium atoms was investigated. In our search we did not observe the conical emission in the far field, although it was previously observed in the case of rubidium. We discuss the possible reason of this unexpected result. The fluorescence spectrum revealed Rb impurity resonance lines in emission due to the collisional redistribution from the K(4p) levels into the Rb(5p) levels. In the forward propagation of 400 nm femtosecond light we observed the molecular band red shifted from potassium second doublet. However, no molecular spectrum was observed when the mode-locked fs laser light was discretely tuned within the wings of the first resonance lines, at 770 nm.

Standoff analysis of laser-produced plasmas using laser-induced fluorescence

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

Harilal, S. S.; Brumfield, B. E.; Phillips, M. C.

We report the use of laser-induced fluorescence (LIF) of laser ablation plumes for standoff applications. The standoff analysis of Al species, as major and minor species in samples, is performed in a nanosecond laser-produced plasma created at a distance ~10 m. The LIF analysis is performed by resonantly exciting an Al transition at 394.4 nm using a continuous wave (cw) tunable laser and by collecting the direct-line fluorescence signal at 396.15 nm. The spectral resolution of LIF is obtained by scanning the cw tunable LIF laser across the selected Al transition. Our results highlight that LIF provides enhanced signal intensity,more » emission persistence, and spectral resolution when compared to thermally-excited emission, and these are crucial considerations for using laser-produced plasma for standoff isotopic analysis.« less

Fluorescence lifetime of normal, benign, and malignant thyroid tissues

NASA Astrophysics Data System (ADS)

Brandao, Mariana; Iwakura, Ricardo; Basilio, Fagne; Haleplian, Kaique; Ito, Amando; de Freitas, Luiz Carlos Conti; Bachmann, Luciano

2015-06-01

Fine-needle aspiration cytology is the standard technique to diagnose thyroid pathologies. However, this method results in a high percentage of inconclusive and false negatives. The use of time-resolved fluorescence techniques to detect biochemical composition and tissue structure alterations could help to develop a portable, minimally invasive, and nondestructive method to assist during surgical procedures. This study aimed to use fluorescence lifetimes to differentiate healthy and benign tissues from malignant thyroid tissue. The thyroid tissue was excited at 298-300 nm and the fluorescence decay registered at 340 and 450 nm. We observed fluorescence lifetimes at 340 nm emission of 0.80±0.26 and 3.94±0.47 ns for healthy tissue; 0.90±0.24 and 4.05±0.46 ns for benign lesions; and 1.21±0.14 and 4.63±0.25 ns for malignant lesions. For 450 nm emissions, we obtain lifetimes of 0.25±0.18 and 3.99±0.39 ns for healthy tissue, 0.24±0.17 and 4.20±0.48 ns for benign lesions, 0.33±0.32 and 4.55±0.55 ns for malignant lesions. Employing analysis of variance, we differentiate malignant lesions from benign and healthy tissues. In addition, we use quadratic discriminant analysis to distinguish malignant from benign and healthy tissues with an accuracy of 76.1%, sensitivity of 74.7%, and specificity of 83.3%. These results indicate that time-resolved fluorescence can assist medical evaluation of thyroid pathologies during surgeries.

The enhanced and broadband near-infrared emission in Pr3+/Nd3+ co-doped tellurite glass

NASA Astrophysics Data System (ADS)

Zhou, Zizhong; Zhou, Yaxun; Cheng, Pan; Zhou, Minghan; Su, Xiue; Li, Jun

2017-11-01

This paper reports an enhanced and broadband near-infrared fluorescence emission in the Pr3+/Nd3+ co-doped tellurite glass, which was prepared using melt-quenching technique. Under the excitation of 488 nm laser diode (LD), three near-infrared emission bands at around 0.9, 1.04 and 1.30 μm from 3P1,0 → 1G4, 1G4→3H4 and 1G4→3H5 radiative transitions respectively were observed in the Pr3+ single-doped glass, and the fluorescence intensities increased further with the introduction of Nd3+ ions, which is mainly attributed to the energy transfers from Nd3+ to Pr3+ emissions. Meanwhile, the spectral overlapping of Pr3+:1G4→3H4 and Nd3+:4F3/2 → 4I11/2 radiative transitions resulted in a broadband emission ranging from 1000 to 1100 nm, whose full-width at half-maximum (FWHM) reached about 66 nm. Additionally, the spectroscopic properties of Nd3+ and Pr3+ ions were analyzed using Judd-Ofelt theory and the thermal stability property of prepared glass was characterized by the differential scanning calorimeter (DSC) measurement, and larger than 134 °C for the difference ΔT(=Tx -Tg) was observed, which indicates its feasibility for later fiber drawing. The enhanced fluorescence and broadband emission indicate that Pr3+/Nd3+ co-doped tellurite glass can be applied in the near-infrared band tunable lasers and broadband optical amplifiers.

Differentiation of cancerous and normal brain tissue using label free fluorescence and Stokes shift spectroscopy

NASA Astrophysics Data System (ADS)

Zhou, Yan; Wang, Leana; Liu, Cheng-hui; He, Yong; Yu, Xinguang; Cheng, Gangge; Wang, Peng; Shu, Cheng; Alfano, Robert R.

2016-03-01

In this report, optical biopsy was applied to diagnose human brain cancer in vitro for the identification of brain cancer from normal tissues by native fluorescence and Stokes shift spectra (SSS). 77 brain specimens including three types of human brain tissues (normal, glioma and brain metastasis of lung cancers) were studied. In order to observe spectral changes of fluorophores via fluorescence, the selected excitation wavelength of UV at 300 and 340 nm for emission spectra and a different Stokes Shift spectra with intervals Δλ = 40 nm were measured. The fluorescence spectra and SSS from multiple key native molecular markers, such as tryptophan, collagen, NADH, alanine, ceroid and lipofuscin were observed in normal and diseased brain tissues. Two diagnostic criteria were established based on the ratios of the peak intensities and peak position in both fluorescence and SSS spectra. It was observed that the ratio of the spectral peak intensity of tryptophan (340 nm) to NADH (440 nm) increased in glioma, meningioma (benign), malignant meninges tumor, and brain metastasis of lung cancer tissues in comparison with normal tissues. The ratio of the SS spectral peak (Δλ = 40 nm) intensities from 292 nm to 366 nm had risen similarly in all grades of tumors.

Fluorescence detection of glutathione and oxidized glutathione in blood with a NIR-excitable cyanine probe

NASA Astrophysics Data System (ADS)

Liu, Chang-hui; Qi, Feng-pei; Wen, Fu-bin; Long, Li-ping; Liu, Ai-juan; Yang, Rong-hua

2018-04-01

Cyanine has been widely utilized as a near infrared (NIR) fluorophore for detection of glutathione (GSH). However, the excitation of most of the reported cyanine-based probes was less than 800 nm, which inevitably induce biological background absorption and lower the sensitivity, limiting their use for detection of GSH in blood samples. To address this issue, here, a heptamethine cyanine probe (DNIR), with a NIR excitation wavelength at 804 nm and a NIR emission wavelength at 832 nm, is employed for the detection of GSH and its oxidized form (GSSG) in blood. The probe displays excellent selectivity for GSH over GSSG and other amino acids, and rapid response to GSH, in particular a good property for indirect detection of GSSG in the presence of enzyme glutathione reductase and the reducing agent nicotinamideadenine dinucleotide phosphate, without further separation prior to fluorescent measurement. To the best of our knowledge, this is the first attempt to explore NIR fluorescent approach for the simultaneous assay of GSH and GSSG in blood. As such, we expect that our fluorescence sensors with both NIR excitation and NIR emission make this strategy suitable for the application in complex physiological systems.

Two-photon excited fluorescence lifetime imaging and spectroscopy of melanins in vitro and in vivo.

PubMed

Krasieva, Tatiana B; Stringari, Chiara; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L; Gratton, Enrico; Tromberg, Bruce J

2013-03-01

Changes in the amounts of cellular eumelanin and pheomelanin have been associated with carcinogenesis. The goal of this work is to develop methods based on two-photon-excited-fluorescence (TPEF) for measuring relative concentrations of these compounds. We acquire TPEF emission spectra (λ(ex)=1000  nm) of melanin in vitro from melanoma cells, hair specimens, and in vivo from healthy volunteers. We find that the pheomelanin emission peaks at approximately 615 to 625 nm and eumelanin exhibits a broad maximum at 640 to 680 nm. Based on these data we define an optical melanin index (OMI) as the ratio of fluorescence intensities at 645 and 615 nm. The measured OMI for the MNT-1 melanoma cell line is 1.6 ± 0.22 while the Mc1R gene knockdown lines MNT-46 and MNT-62 show substantially greater pheomelanin production (OMI=0.5 ± 0.05 and 0.17 ± 0.03, respectively). The measured values are in good agreement with chemistry-based melanin extraction methods. In order to better separate melanin fluorescence from other intrinsic fluorophores, we perform fluorescence lifetime imaging microscopy of in vitro specimens. The relative concentrations of keratin, eumelanin, and pheomelanin components are resolved using a phasor approach for analyzing lifetime data. Our results suggest that a noninvasive TPEF index based on spectra and lifetime could potentially be used for rapid melanin ratio characterization both in vitro and in vivo.

Two-photon excited fluorescence lifetime imaging and spectroscopy of melanins in vitro and in vivo

NASA Astrophysics Data System (ADS)

Krasieva, Tatiana B.; Stringari, Chiara; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Gratton, Enrico; Tromberg, Bruce J.

2013-03-01

Changes in the amounts of cellular eumelanin and pheomelanin have been associated with carcinogenesis. The goal of this work is to develop methods based on two-photon-excited-fluorescence (TPEF) for measuring relative concentrations of these compounds. We acquire TPEF emission spectra (λex=1000 nm) of melanin in vitro from melanoma cells, hair specimens, and in vivo from healthy volunteers. We find that the pheomelanin emission peaks at approximately 615 to 625 nm and eumelanin exhibits a broad maximum at 640 to 680 nm. Based on these data we define an optical melanin index (OMI) as the ratio of fluorescence intensities at 645 and 615 nm. The measured OMI for the MNT-1 melanoma cell line is 1.6±0.22 while the Mc1R gene knockdown lines MNT-46 and MNT-62 show substantially greater pheomelanin production (OMI=0.5±0.05 and 0.17±0.03, respectively). The measured values are in good agreement with chemistry-based melanin extraction methods. In order to better separate melanin fluorescence from other intrinsic fluorophores, we perform fluorescence lifetime imaging microscopy of in vitro specimens. The relative concentrations of keratin, eumelanin, and pheomelanin components are resolved using a phasor approach for analyzing lifetime data. Our results suggest that a noninvasive TPEF index based on spectra and lifetime could potentially be used for rapid melanin ratio characterization both in vitro and in vivo.

Two-photon excited fluorescence lifetime imaging and spectroscopy of melanins in vitro and in vivo

PubMed Central

Krasieva, Tatiana B.; Stringari, Chiara; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Gratton, Enrico

2012-01-01

Abstract. Changes in the amounts of cellular eumelanin and pheomelanin have been associated with carcinogenesis. The goal of this work is to develop methods based on two-photon-excited-fluorescence (TPEF) for measuring relative concentrations of these compounds. We acquire TPEF emission spectra (λex=1000  nm) of melanin in vitro from melanoma cells, hair specimens, and in vivo from healthy volunteers. We find that the pheomelanin emission peaks at approximately 615 to 625 nm and eumelanin exhibits a broad maximum at 640 to 680 nm. Based on these data we define an optical melanin index (OMI) as the ratio of fluorescence intensities at 645 and 615 nm. The measured OMI for the MNT-1 melanoma cell line is 1.6±0.22 while the Mc1R gene knockdown lines MNT-46 and MNT-62 show substantially greater pheomelanin production (OMI=0.5±0.05 and 0.17±0.03, respectively). The measured values are in good agreement with chemistry-based melanin extraction methods. In order to better separate melanin fluorescence from other intrinsic fluorophores, we perform fluorescence lifetime imaging microscopy of in vitro specimens. The relative concentrations of keratin, eumelanin, and pheomelanin components are resolved using a phasor approach for analyzing lifetime data. Our results suggest that a noninvasive TPEF index based on spectra and lifetime could potentially be used for rapid melanin ratio characterization both in vitro and in vivo. PMID:23235925

ATOTA-a very promising green fluorophore

NASA Astrophysics Data System (ADS)

Doan, Hung The

Despite the fact that fluorescence community nowadays has invested in developing near-infrared probes, green fluorescence dyes like fluorescein and substitutes are still among the most widely used fluorophores for labeling in cellular imaging and biomedical research. Trioxatriangulenium dye ATOTA + is a very promising green fluorophore with high extinction coefficient and outstanding fluorescence quantum yield. This study focuses on characterizing ATOTA+'s fundamental spectroscopic properties, including fluorescence and orientation of the transition moments. ATOTA's aggregation in aqueous solution and lipid bilayer membrane are also investigated. ATOTA+ has absorption maxima between 470 nm and 476 nm and emission maxima between 496 nm and 511 nm depending on the solvent. The molar extinction coefficient varies from 135,000 mol-1cm-1 in nonpolar dichloromethane to above 90,000 mol-1cm-1 in polar solvents such as methanol. The quantum yield of ATOTA+ is close to 1 in nonpolar DCM and decreases to 0.44 in polar DMF. ATOTA+'s fluorescence lifetimes vary between 3.25 ns in aprotic low polarity triacetin to 1.66 ns in polar DMF. Furthermore, both radiative and non-radiative rates are affected by solvent polarity. ATOTA+ has very low water solubility due to the presence of 6 diethyl substitutions, and forms H-aggregates with a blue-shifted absorption maxima around 450 nm and red-shifted emission maxima of 580 nm respectively with fluorescence lifetime above 20 ns. The excitation anisotropy approaches 0.35 at red edge of the absorption spectrum and shape of polarization spectrum suggests the presence of overlapping transition moments in a S0-S1 band which is confirmed by linear dichroism in stretched PVA film. In DMPC lipid vesicles, ATOTA + forms a tight ion pair with a counter anion and localizes in the hydrocarbon interior. Overall we conclude that ATOTA+ will be a highly useful and superior member of the green fluorophore family.

Selective colorimetric and fluorescent quenching determination of uranyl ion via its complexation with curcumin

NASA Astrophysics Data System (ADS)

Zhu, Jing-Hui; Zhao, Xin; Yang, Jidong; Tan, Yu-Ting; Zhang, Lei; Liu, Shao-Pu; Liu, Zhong-Fang; Hu, Xiao-Li

2016-04-01

Under pH 4.0 HAc-NaAc buffer medium, curcumin alone possesses extraordinary weak fluorescence emission. Nevertheless, the introduction of Triton X-100 micelles can largely enhance the fluorescence intensity of curcumin. Uranyl ions can complex with micelles-capped curcumin, along with the slight red shift of curcumin fluorescence (about 1-7 nm), a clear decrement of absorbance (424 nm) and fluorescence (507 nm) intensities, and a distinct color change from bright yellow to orange. The fluorescence decrements (ΔF, 507 nm) are positively correlated to the amount of uranyl ions in the concentration range of 3.7 × 10- 6-1.4 × 10- 5 mol L- 1. The detection limit of this fluorescence quenching methods is 3.7 × 10- 6 mol L- 1, which is nearly 9000 times lower than the maximum allowable level in drinking water proposed by World Health Organization. Good selectivity is achieved because of a majority of co-existing substances (such as Ce4 +, La3 +, and Th4 +) do not affect the detection. The content of uranyl ions in tap water samples was determined by the proposed method with satisfactory results.

Extended wavelength anisotropy resolved multidimensional emission spectroscopy (ARMES) measurements: better filters, validation standards, and Rayleigh scatter removal methods

NASA Astrophysics Data System (ADS)

Casamayou-Boucau, Yannick; Ryder, Alan G.

2017-09-01

Anisotropy resolved multidimensional emission spectroscopy (ARMES) provides valuable insights into multi-fluorophore proteins (Groza et al 2015 Anal. Chim. Acta 886 133-42). Fluorescence anisotropy adds to the multidimensional fluorescence dataset information about the physical size of the fluorophores and/or the rigidity of the surrounding micro-environment. The first ARMES studies used standard thin film polarizers (TFP) that had negligible transmission between 250 and 290 nm, preventing accurate measurement of intrinsic protein fluorescence from tyrosine and tryptophan. Replacing TFP with pairs of broadband wire grid polarizers enabled standard fluorescence spectrometers to accurately measure anisotropies between 250 and 300 nm, which was validated with solutions of perylene in the UV and Erythrosin B and Phloxine B in the visible. In all cases, anisotropies were accurate to better than ±1% when compared to literature measurements made with Glan Thompson or TFP polarizers. Better dual wire grid polarizer UV transmittance and the use of excitation-emission matrix measurements for ARMES required complete Rayleigh scatter elimination. This was achieved by chemometric modelling rather than classical interpolation, which enabled the acquisition of pure anisotropy patterns over wider spectral ranges. In combination, these three improvements permit the accurate implementation of ARMES for studying intrinsic protein fluorescence.

Laser Induced Fluorescence of Helium Ions in a Helicon Plasma

NASA Astrophysics Data System (ADS)

Compton, C. S.; Biloui, C.; Hardin, R. A.; Keesee, A. M.; Scime, E. E.; Boivin, R.

2003-10-01

The lack of a suitable Laser Induced Fluorescence (LIF) scheme for helium ions at visible wavelengths has prevented LIF from being employed in helium plasmas for measurements of ion temperature and bulk ion flow speeds. In this work, we will discuss our attempts to perform LIF of helium ions in a helicon source plasma using an infrared, tunable diode laser operating at 1012.36 nm. The infrared transition corresponds to excitation from the n = 4 level (4f ^2F) to the n = 5 (5g ^2G) level of singly ionized helium and therefore requires substantial electron temperatures (> 10 eV) to maintain an adequate ion population in the n = 4 state. Calculations using a steady state coronal model predict that the n = 4 state population will be 25% larger than the n = 5 population for our experimental conditions. The fluorescence decay from the n = 5 (5f ^2F) level of singly ionized helium level to the n = 3 (3d ^2D) level at 320.31 nm is monitored as the diode laser is swept through 10 GHz around the 1012.36 nm line. Note that the fluorescence emission requires a collisionally coupled transition between two different n = 5 quantum states. We will also present measurements of the emission intensities of both the 1012.36 nm and the 320.31 nm lines as a function of source neutral pressure, rf power, and plasma density. This work supported by the U.S. DoE EPSCoR Lab Partnership Program.

Optical Temperature Sensor Based on Infrared Excited Green Upconversion Emission in Hexagonal Phase NaLuF4:Yb3+/Er3+ Nanorods.

PubMed

Li, Dongyu; Tian, Linlin; Huang, Zhen; Shao, Lexi; Quan, Jun; Wang, Yuxiao

2016-04-01

Hexagonal phase NaLuF4:Yb3+/Er3+ nanorods were synthesized hydrothermally. An analysis of the intense green upconversion emissions at 525 nm and 550 nm in hexagonal phase NaLuF4:Yb3/+Er3+ nanorods under excitation power density of 4.2 W/cm2 available from a diode laser emitting at 976 nm, have been undertaken. Fluorescence intensity ratio (FIR) variation of temperature-sensitive green upconversion emissions at 525 nm and 550 nm in this material was recorded in the physiological range from 295 to 343 K. The maximum sensitivity derived from the FIR technique of the green upconversion emissions is approximately 0.0044 K-1. Experimental results implied that hexagonal phase NaLuF4:Yb3/+Er3+ nanorods was a potential candidate for optical temperature sensor.

Capacitively Coupled Plasma Discharge of Ionic Liquid Solutions to Synthesize Carbon Dots as Fluorescent Sensors.

PubMed

Ke, Ching-Bin; Lu, Te-Ling; Chen, Jian-Lian

2018-05-26

Oxygen and nitrogen capacitively coupled plasma (CCP) was used to irradiate mixtures of aliphatic acids in high boiling point solvents to synthesize fluorescent carbon dots (C-dots). With a high fluorescence intensity, the C-dots obtained from the O₂/CCP radiation of a 1-ethyl-3-methylimidazolium dicyanamide ionic liquid solution of citric acid were characterized with an average diameter of 8.6 nm (σ = 1.1 nm), nitrogen and oxygen bonding functionalities, excitation-independent emissions, and upconversion fluorescence. Through dialysis of the CCP-treated C-dots, two emissive surface states corresponding to their respective functionalities and emissions were identified. The fluorescence spectrum of the CCP-treated C-dots was different from that of the microwave irradiation and possessed higher intensity than that of hydrothermal pyrolysis. By evaluation of the fluorescence quenching effect on flavonoids and metal ions, the CCP-treated C-dots showed a high selectivity for quercetin and sensitivity to Hg 2+ . Based on the Perrin model, a calibration curve ( R ² = 0.9992) was established for quercetin ranging from 2.4 μM to 119 μM with an LOD (limit of detection) = 0.5 μM. The quercetin in the ethanol extract of the sun-dried peel of Citrus reticulata cv. Chachiensis was determined by a standard addition method to be 4.20 ± 0.15 mg/g with a matrix effect of 8.16%.

Optical measurement of acidification of human dental plaque in vitro

NASA Astrophysics Data System (ADS)

Graham, Jasmine Y.; Nelson, Leonard Y.; Seibel, Eric J.

2018-02-01

A pH measurement of oral biofilms is helpful for monitoring the impact of acidogenic bacteria in the caries process. Demineralization of dental enamel is closely related to the time dependent pH of human plaque. Therefore, providing a means to easily measure the local pH of biofilms is a useful clinical diagnostic in the arsenal of caries prevention tools. Optical measurement methods of plaque metabolism can use intrinsic fluorescence or extrinsic fluorescence from added dyes. Autofluorescence spectral features of human oral biofilms at green (500 nm) and red (634 nm) fluorescence wavelengths using 405 nm excitation did not demonstrate a spectral or intensity shift between neutral and acidic conditions. Chlorin e6, an ingredient in chlorophyllin food supplement, exhibited a spectral and intensity shift of fluorescence emission in buffered solutions, but this quantitative pH-dependence was not transferable to a human plaque environment. Finally, a ratiometric quantitative pH measure was achieved by exciting (405 nm laser) a mixture of two dyes, fluorescein and rhodamine B. This two-dye mixture produced two strong fluorescent bands centered at 515 nm (fluorescein) and 580 nm (rhodamine B), where the 515 nm band was pH sensitive and the 580 nm band served as a pH insensitive reference. This dual-dye fluorescence ratio exhibited a linear response over pH 7 to 5 in human oral biofilms during a sugar challenge. We have explored methods to use non-contact, optical measures of local acidity levels in difficult to access dental locations such as occlusal fissures using various pH sensitive fluorescent dye systems.

Detection of iron atoms by emission spectroscopy and laser-induced fluorescence in solid propellant flames.

PubMed

Vilmart, G; Dorval, N; Orain, M; Lambert, D; Devillers, R; Fabignon, Y; Attal-Tretout, B; Bresson, A

2018-05-10

Planar laser-induced fluorescence on atomic iron is investigated in this paper, and a measurement strategy is proposed to monitor the fluorescence of iron atoms with good sensitivity. A model is proposed to fit the experimental fluorescence spectra, and good agreement is found between simulated and experimental spectra. Emission and laser-induced fluorescence measurements are performed in the flames of ammonium perchlorate composite propellants containing iron-based catalysts. A fluorescence signal from iron atoms after excitation at 248 nm is observed for the first time in propellant flames. Images of the spatial distribution of iron atoms are recorded in the flame in which turbulent structures are generated. Iron fluorescence is detected up to 1.0 MPa, which opens the way to application in propellant combustion.

Long wave fluorophore sensor compounds and other fluorescent sensor compounds in polymers

DOEpatents

Walsh, Joseph C.; Heiss, Aaron M.; Noronha, Glenn; Vachon, David J.; Lane, Stephen M.; Satcher, Jr., Joe H.; Peyser, Thomas A.; Van Antwerp, William Peter; Mastrototaro, John Joseph

2004-07-20

Fluorescent biosensor molecules, fluorescent biosensors and systems, as well as methods of making and using these biosensor molecules and systems are described. Embodiments of these biosensor molecules exhibit fluorescence emission at wavelengths greater than about 650 nm. Typical biosensor molecules include a fluorophore that includes an iminium ion, a linker moiety that includes a group that is an anilinic type of relationship to the fluorophore and a boronate substrate recognition/binding moiety, which binds glucose. The fluorescence molecules modulated by the presence or absence of polyhydroxylated analytes such as glucose. This property of these molecules of the invention, as well as their ability to emit fluorescent light at greater than about 650 nm, renders these biosensor molecules particularly well-suited for detecting and measuring in-vivo glucose concentrations.

Enhancing early bladder cancer detection with fluorescence-guided endoscopic optical coherence tomography

NASA Astrophysics Data System (ADS)

Pan, Y. T.; Xie, T. Q.; Du, C. W.; Bastacky, S.; Meyers, S.; Zeidel, M. L.

2003-12-01

We report an experimental study of the possibility of enhancing early bladder cancer diagnosis with fluorescence-image-guided endoscopic optical coherence tomography (OCT). After the intravesical instillation of a 10% solution of 5-aminolevulinic acid, simultaneous fluorescence imaging (excitation of 380-420 nm, emission of 620-700 nm) and OCT are performed on rat bladders to identify the photochemical and morphological changes associated with uroepithelial tumorigenesis. The preliminary results of our ex vivo study reveal that both fluorescence and OCT can identify early uroepithelial cancers, and OCT can detect precancerous lesions (e.g., hyperplasia) that fluorescence may miss. This suggests that a cystoscope combining 5-aminolevulinic acid fluorescence and OCT imaging has the potential to enhance the efficiency and sensitivity of early bladder cancer diagnosis.

Measurement of Fluorescence in a Rhodamine-123 Doped Self-Assembled “Giant” Mesostructured Silica Sphere Using a Smartphone as Optical Hardware

PubMed Central

Canning, John; Lau, Angelica; Naqshbandi, Masood; Petermann, Ingemar; Crossley, Maxwell J.

2011-01-01

The blue OLED emission from a mobile phone was characterised, revealing a sharp emission band centred at λ = 445 nm with a 3dB bandwidth Δλ ∼ 20 nm. It was used to excite Rhodamine 123 doped within a “giant” mesostructured silica sphere during fabrication through evaporative self-assembly of silica nanoparticles. Fluorescence was able to be detected using a standard optical microscope fitted with a green transmission pass filter and cooled CCD and with 1 ms exposure time demonstrating the potential of mobile platforms as the basis for portable diagnostics in the field. PMID:22164002

Diffuse reflectance of the ocean - The theory of its augmentation by chlorophyll a fluorescence at 685 nm

NASA Technical Reports Server (NTRS)

Gordon, H. R.

1979-01-01

The radiative transfer equation is modified to include the effect of fluorescent substances and solved in the quasi-single scattering approximation for a homogeneous ocean containing fluorescent particles with wavelength independent quantum efficiency and a Gaussian shaped emission line. The results are applied to the in vivo fluorescence of chlorophyll a (in phytoplankton) in the ocean to determine if the observed quantum efficiencies are large enough to explain the enhancement of the ocean's diffuse reflectance near 685 nm in chlorophyll rich waters without resorting to anomalous dispersion. The computations indicate that the required efficiencies are sufficiently low to account completely for the enhanced reflectance. The validity of the theory is further demonstrated by deriving values for the upwelling irradiance attenuation coefficient at 685 nm which are in close agreement with the observations.

Masked red-emitting carbopyronine dyes with photosensitive 2-diazo-1-indanone caging group.

PubMed

Kolmakov, Kirill; Wurm, Christian; Sednev, Maksim V; Bossi, Mariano L; Belov, Vladimir N; Hell, Stefan W

2012-03-01

Caged near-IR emitting fluorescent dyes are in high demand in optical microscopy but up to now were unavailable. We discovered that the combination of a carbopyronine dye core and a photosensitive 2-diazo-1-indanone residue leads to masked near-IR emitting fluorescent dyes. Illumination of these caged dyes with either UV or visible light (λ < 420 nm) efficiently generates fluorescent compounds with absorption and emission at 635 nm and 660 nm, respectively. A high-yielding synthetic route with attractive possibilities for further dye design is described in detail. Good photostability, high contrast, and a large fluorescence quantum yield after uncaging are the most important features of the new compounds for non-invasive imaging in high-resolution optical microscopy. For use in immunolabelling the caged dyes were decorated with a (hydrophilic) linker and an (activated) carboxyl group.

Enhancement of 800 nm upconversion emission in a thulium doped tellurite microstructured fiber pumped by a 1560 nm femtosecond fiber laser

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

Jia, Zhixu; Zheng, Kezhi; State Key Laboratory on Integrated Optoelectronics, College of Electronic Science and Engineering, Jilin University, Changchun 130012

We report enhanced upconversion (UC) fluorescence in Tm{sup 3+} doped tellurite microstructured fibers (TDTMFs) fabricated by using a rod-in-tube method. Under the pumping of a 1560 nm femtosecond fiber laser, ultrabroadband supercontinuum light expanding from ∼1050 to ∼2700 nm was generated in a 4 cm long TDTMF. Simultaneously, intense 800 nm UC emission from the {sup 3}H{sub 4} → {sup 3}H{sub 6} transition of Tm{sup 3+} was observed in the same TDTMF. Compared to that pumped by a 1560 nm continuous wave fiber laser, the UC emission intensity was enhanced by ∼4.1 times. The enhancement was due to the spectral broadening in the TDTMF under themore » pumping of the 1560 nm femtosecond fiber laser.« less

Solid-state phosphorescence-to-fluorescence switching in a cyclometalated Ir(III) complex containing an acid-labile chromophoric ancillary ligand: implication for multimodal security printing.

PubMed

Whang, Dong Ryeol; You, Youngmin; Chae, Weon-Sik; Heo, Jeongyun; Kim, Sehoon; Park, Soo Young

2012-11-06

In this study, we have demonstrated the reconstruction of encrypted information by employing photoluminescence spectra and lifetimes of a phosphorescent Ir(III) complex (IrHBT). IrHBT was constructed on the basis of a heteroleptic structure comprising a fluorescent N^O ancillary ligand. From the viewpoint of information security, the transformation of the Ir(III) complex between phosphorescent and fluorescent states can be encoded with chemical/photoirradiation methods. Thin polymer films (poly(methylmethacrylate), PMMA) doped with IrHBT display long-lived emission typical of phosphorescence (λ(max) = 586 nm, τ(obs) = 2.90 μs). Meanwhile, exposure to HCl vapor switches the emission to fluorescence (λ(max) = 514 nm, τ(obs) = 1.53 ns) with drastic changes in both the photoluminescence color and lifetime. Security printing on paper impregnated with IrHBT or on a PMMA film containing IrHBT and photoacid generator (triphenylsulfonium triflate) enables the bimodal readout of photoluminescence color and lifetime.

A chromogenic and fluorogenic rhodol-based chemosensor for hydrazine detection and its application in live cell bioimaging

NASA Astrophysics Data System (ADS)

Tiensomjitr, Khomsan; Noorat, Rattha; Chomngam, Sinchai; Wechakorn, Kanokorn; Prabpai, Samran; Kanjanasirirat, Phongthon; Pewkliang, Yongyut; Borwornpinyo, Suparerk; Kongsaeree, Palangpon

2018-04-01

A rhodol-based fluorescent probe has been developed as a selective hydrazine chemosensor using levulinate as a recognition site. The rhodol levulinate probe (RL) demonstrated high selectivity and sensitivity toward hydrazine among other molecules. The chromogenic response of RL solution to hydrazine from colorless to pink could be readily observed by the naked eye, while strong fluorescence emission could be monitored upon excitation at 525 nm. The detection process occurred via a ring-opening process of the spirolactone initiated by hydrazinolysis, triggering the fluorescence emission with a 53-fold enhancement. The probe rapidly reacted with hydrazine in aqueous medium with the detection limit of 26 nM (0.83 ppb), lower than the threshold limit value (TLV) of 10 ppb suggested by the U.S. Environmental Protection Agency. Furthermore, RL-impregnated paper strips could detect hydrazine vapor. For biological applicability of RL, its membrane-permeable property led to bioimaging of hydrazine in live HepG2 cells by confocal fluorescence microscopy.

One-step aqueous synthesis of fluorescent copper nanoclusters by direct metal reduction

NASA Astrophysics Data System (ADS)

Fernández-Ujados, Mónica; Trapiella-Alfonso, Laura; Costa-Fernández, José M.; Pereiro, Rosario; Sanz-Medel, Alfredo

2013-12-01

A one-step aqueous synthesis of highly fluorescent water-soluble copper nanoclusters (CuNCs) is here described, based on direct reduction of the metal precursor with NaBH4 in the presence of bidentate ligands (made of lipoic acid anchoring groups, appended with a poly(ethylene glycol) short chain). A complete optical and structural characterization was carried out: the optical emission was centred at 416 nm, with a luminescence quantum yield in water of 3.6% (the highest one reported so far in water for this kind of nanocluster). The structural characterization reveals a homogeneous size distribution (of 2.5 nm diameter) with spherical shape. The CuNCs obtained offer long-term stability (the luminescence emission remained unaltered after more than two months) under a broad range of chemical conditions (e.g. stored at pH 3-12 or even in a high ionic strength medium such as 1 M NaCl) and high photostability, keeping their fluorescence emission intact after more than 2 h of daylight and UV-light exposition. All those advantageous features warrant synthesized CuNCs being promising fluorescent nanoprobes for further developments including (bio)applications.

Using violet laser-induced chlorophyll fluorescence emission spectra for crop yield assessment of cowpea (Vigna unguiculata (L) Walp) varieties

NASA Astrophysics Data System (ADS)

Anderson, Benjamin; Buah-Bassuah, Paul K.; Tetteh, Jonathan P.

2004-07-01

The use of violet laser-induced chlorophyll fluorescence (LICF) emission spectra to monitor the growth of five varieties of cowpea in the University of Cape Coast Botanical Garden is presented. Radiation from a continuous-wave violet laser diode emitting at 396 nm through a fibre is closely incident on in vivo leaves of cowpea to excite chlorophyll fluorescence, which is detected by an integrated spectrometer with CCD readout. The chlorophyll fluorescence spectra with peaks at 683 and 731 nm were used for growth monitoring of the cowpea plants over three weeks and analysed using Gaussian spectral functions with curve fitted parameters to determine the peak positions, area under the spectral curve and the intensity ratio F683/F731. The variation in the intensity ratio of the chlorophyll bands showed sensitive changes indicating the photosynthetic activity of the cowpea varieties. A discussion of the fluorescence result as compared to conventional assessment is presented with regard to discrimination between the cowpea varieties in terms of crop yield performance.

Fluorescence lifetime imaging microscopy using near-infrared contrast agents.

PubMed

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

2012-08-01

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

Fluorescence Lifetime Imaging Microscopy Using Near-Infrared Contrast Agents

PubMed Central

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

2013-01-01

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

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

PubMed Central

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

2014-01-01

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

Interferences of resveratrol with fura-2-derived fluorescence in intracellular free-Ca(2+) concentration determinations.

PubMed

Santofimia-Castaño, Patricia; Salido, Gines M; Gonzalez, Antonio

2016-08-01

Resveratrol (3,4',5-trihydroxy-trans-stilbene) is an antioxidant widely employed in cell physiology studies. It has been reported that it interferes with fura-2-derived fluorescence, making the employment of this dye nonviable. In this work, the interference of resveratrol with fura-2 determinations of intracellular free-Ca(2+) concentration ([Ca(2+)]c) was examined. Solutions containing different concentrations of resveratrol, with or without fura-2, in the presence or in the absence of Ca(2+), were analyzed by spectrofluorimetry. AR42J tumor cells were employed to study the influence of resveratrol on fura-2 fluorescence in living cells, by single cell fluorimetry. Resveratrol impaired the detection of fura-2-fluorescence emission (510 nm) at the 340, 360 and 380 nm excitation wavelengths. Resveratrol emitted fluorescence at 510 nm when lighted at all three excitation wavelengths. In addition, resveratrol emitted fluorescence at 380 nm when excited at 340 nm. Our observations suggest that the employment of the ratiometric properties of fura-2 to follow changes in [Ca(2+)]c in the presence of resveratrol is not viable. However, we think that the 380 nm excitation light could be employed. Results could be expressed as F0/F380, where F0 is the resting fluorescence and F380 is the value of fluoresce at a certain time point. We could follow changes in [Ca(2+)]c evoked by CCK-8, and we also detected Ca(2+) mobilization by 100 µM resveratrol in AR42J cells. This investigation presents evidence demonstrating that resveratrol interferes with fura-2 fluorescence spectra. Nevertheless, a chance still exists if the 380 nm excitation wavelength is employed in the middle or low micromolar concentrations of resveratrol.

[A new class of exciplex-formed probe detect of specific sequence DNA].

PubMed

Li, Qing-Yong; Zu, Yuan-Gang; Lü, Hong-Yan; Wang, Li-Min

2009-07-01

The present research was to develop the exciplex-based fluorescence detection of DNA. A SNP-containing region of cytochrome P450 2C9 DNA systems was evaluated to define some of the structural and associated requirement of this new class of exciplex-formed probe, and a 24-base target was selected which contains single-nucleotide polymorphisms (SNP) in genes coding for cytochrome P450. The two probes were all 12-base to give coverage of a 24-base target region to ensure specificity within the human genome. Exciplex partners used in this study were prepared using analogous phosphoramide attachment to the 3'- or 5'-phosphate group of the appropriate oligonucleotide probes. The target effectively assembled its own detector by hybridization from components which were non-fluorescent at the detection wavelength, leading to the huge improvement in terms of decreased background. This research provides details of the effects of different partner, position of partners and different excitation wavelengths for the split-oligonucleotide probe system for exciplex-based fluorescence detection of DNA. This study demonstrates that the emission intensity of the excimer formed by new pyrene derivative is the highest in these excimer and exciplex, and the excimer is easy to be formed and not sensitive to the position of partners. However the exciplex formed by the new pyrene derivative and naphthalene emitted strongly at -505 nm with large Stokes shifts (120-130 nm), and the monomer emission at 390 and 410 nm is nearly zero. Excitation wavelength of 400 nm is the best for I(e505)/I(m410) (exciplex emission at 505 nm/monomer emission at 410 nm) of the exciplex. This method features lower background and high sensitivity. Moreover the exciplex is sensitive to the steric factor, different position of partners and microenvironment, so this exciplex system is promising and could be tried to identify the SNP genes.

Use of the fluorescence quantum yield for the determination of the number-average molecular weight of polymers of epicatechin with 4β→8 interflavin bonds

Treesearch

D. Cho; W.L. Mattice; L.J. Porter; Richard W. Hemingway

1989-01-01

Excitation at 280 nm produces a structureless emission band with a maximum at 321-324 nm for dilute solutions of catechin, epicatechin, and their oligomers in l,4-dioxane or water. The fluorescence quantum yield, Q, has been measured in these two solvents for five dimers, a trimer, a tetramer, a pentamer, a hexamer, and a polymer in which the monomer...

Laser-induced fluorescence at 488 nm excitation for detecting benign and malignant lesions in stomach mucosa.

PubMed

Silveira, Landulfo; Betiol Filho, João Angelo; Silveira, Fabricio Luiz; Zângaro, Renato Amaro; Pacheco, Marcos Tadeu T

2008-01-01

This work aims the detection of the histopathologic alterations of in vitro human gastric mucosa using spectral informations from laser-induced fluorescence spectroscopy (LIFS) technique with excitation at 488 nm (argon laser). A total of 108 biopsies with endoscopic diagnosis of gastritis and gastric cancer were obtained at the antral gastric region, from 35 patients with dyspeptic digestive complaints. The biopsies were collected during the endoscopic examination. On each biopsy fragment the autofluorescence spectrum was collected in two random points, through a fiber-optic catheter coupled to the excitation laser. The fluorescence emission spectra collected by the fibers were directed to the spectrograph and detected by the CCD camera. The spectra were then separated in groups (N, normal; LI, light inflammation; MI, moderated inflammation; CA, adenocarcinoma), based on the histopathology. The ratio between the emission wavelengths 550 and 600 nm was used as a diagnostic parameter. Analysis of fluorescence spectra was able to identify the normal tissue from adenocarcinoma lesions with 100% of sensibility and specificity. The ratio intensities between inflammation (light and moderated), although presented significantly statistical differences when compared to the normal mucosa, do not furnish enough sensibility and specificity for use as an identification method due to high variations. LIFS, with excitation of 488 nm, could be used in the differentiation of normal tissue and neoplasic lesions, assisting a less invasive diagnosis.

Picosecond rotationally resolved stimulated emission pumping spectroscopy of nitric oxide

NASA Astrophysics Data System (ADS)

Tanjaroon, Chakree; Reeve, Scott W.; Ford, Alan; Murry, W. Dean; Lyon, Kevin; Yount, Bret; Britton, Dan; Burns, William A.; Allen, Susan D.; Bruce Johnson, J.

2012-01-01

Stimulated emission pumping (SEP) experiments were performed on the nitric oxide molecule in a flow cell environment using lasers with pulse widths of 17-25 ps. A lambda excitation scheme, or ''pump-dump" arrangement, was employed with the pump laser tuned to the T 00 vibronic band origin ( λ=226.35(1)nm) of the A2Σ+( v' = 0, J') ← X2Π1/2( v″ = 0, J″) and the dump laser scanned from 246-248 nm within the A2Σ+( v' = 0, J') → X2Π1/2( v″ = 2, J″) transition. The rotationally resolved SEP spectra were measured by observing the total fluorescence within the A2Σ+( v' = 0, J') → X2Π1/2( v″ = 1, J″) transition between 235 nm and 237.2 nm while scanning the dump laser wavelengths. Multiple rotational states were excited due to the broad laser bandwidth. Measurements showed that the resolved rotational structure depended on the energy and bandwidth of the applied pump and dump laser pulses. Analysis of the observed fluorescence depletion signals yielded an average percent fluorescence depletion of about 19% when λ=226.35(1)nm and λ=247.91(1)nm. This value reflects the percent transfer of the NO population from the A2Σ+( V' = 0, J') excited electronic state to the X2Π1/2( v″ = 2, J″) ground electronic state. The maximum expected depletion is 50% in the limit of dump saturation. Selective excitation of NO at the bandhead provides good spectral discrimination from the background emission and noise and unambiguously confirms the identity of the emitter.

Growth and optical properties of Co,Nd:LaMgAl11O19

NASA Astrophysics Data System (ADS)

Xu, Peng; Xia, Changtai; Di, Juqing; Xu, Xiaodong; Sai, Qinglin; Wang, Lulu

2012-12-01

Nd,Co:LaMgAl11O19 (abbreviated as Co,Nd:LMA) was grown using the Czochralski method. The structure, polarized absorption spectrum, fluorescence spectrum, and fluorescence decay time were analyzed. The as-grown crystal had very wide absorption bands at 794 nm, which can be pumped by GaAs laser diode without temperature stabilization. Two strong emission bands were present at 1056 nm and 1082 nm with full-width at half-maximum (FWHM) of 6 and 8 nm, respectively. The large FWHM is due to the inhomogeneity of the Nd ion sites. The lifetimes of the 4F3/2 manifold of Co,Nd:LMA at room temperature monitored at 905 nm, 1056 nm, and 1344 nm were 292, 288, and 350 μs, respectively, which was caused by the different contribution of the three different sites with D3h and C2v symmetry. The absorption band of Co is from 1.3 μm to 1.6 μm, and Co,Nd:LMA still has a strong emission around the 1.38 μm, indicating that the Co,Nd:LMA can be applied as a potential self-Q-switched material operating at 1.3 μm.

Mercury speciation with fluorescent gold nanocluster as a probe.

PubMed

Yang, Jian-Yu; Yang, Ting; Wang, Xiao-Yan; Chen, Ming-Li; Yu, Yong-Liang; Wang, Jian-Hua

2018-05-11

Fluorescent nanoparticles are widely used for sensing biologically significant species. However, it is rarely reported for the discrimination or speciation of metal species. In this work, we report for the first time the speciation of mercury (Hg 2+ ) and methylmercury (CH 3 Hg + ) by taking advantage of the fluorescence feature of folic acid-capped gold nanoclusters (FA-AuNCs). FA-Au NCs exhibit an average size of 2.08±0.15 nm and a maximum emission at λ ex /λ em = 280/440 nm with a quantum yield of 27.3%. It is interesting that Hg 2+ causes a significant quench on the fluorescence of FA-Au NCs, whereas CH 3 Hg + leads to a remarkable fluorescence enhancement. Based on this discriminative fluorescent response between Hg 2+ and CH 3 Hg + , a novel nanosensor for the speciation of CH 3 Hg + and Hg 2+ was developed, providing limits of detection (LOD) of 28 nM for Hg 2+ and 25 nM for CH 3 Hg + within 100-1000 nM. This sensing system is highly selective to mercury. Its practical applications were further demonstrated by the analysis of CH 3 Hg + and the speciation of mercury (CH 3 Hg + and Hg 2+ ) in environmental water and fish samples.

Detection of intaoral lesions using a fluorescence camera

NASA Astrophysics Data System (ADS)

Thoms, Michael

2006-02-01

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

Synthesis, Luminescent Properties of aza-Boron-Diquinomethene Difluoride Complexes and Their Application for Fluorescent Security Inks.

PubMed

Gu, Long; Liu, Rui; Shi, Hong; Wang, Qiang; Song, Guangliang; Zhu, Xiaolin; Yuan, Shidong; Zhu, Hongjun

2016-03-01

Two aza-boron-diquinomethene (aza-BODIQU) complexes bearing phenyl and carbazyl substituents were synthesized and characterized. Their photophysical properties were investigated systematically via spectroscopic and theoretical methods. Both complexes exhibit strong (1)π-π* transition absorptions (λ(abs) = 400-540 nm) and intense fluorescent emissions (λ(em) = 440-600 nm, Φ(PL) = 0.93 and 0.78) in CH2Cl2 solution and in solid state at room temperature. Compared to the complex with phenyl groups, the complex bearing carbazyl groups shows significant bathochromic shift in both absorption and emission. This could be attributed to the larger π-electron conjugation of the carbazole unit and intramolecular charge transfer feature from carbazole to aza-BODIQU component. In addition, the complexes exhibit intense photoluminescence and good stability on antacid, anti-alkali and stability in printing ink samples, which makes them potential dopants for the application of fluorescent security inks.

Synthesis and spectroscopic characterization of fluorescent 4-aminoantipyrine analogues: Molecular docking and in vitro cytotoxicity studies

NASA Astrophysics Data System (ADS)

Premnath, D.; Mosae Selvakumar, P.; Ravichandiran, P.; Tamil Selvan, G.; Indiraleka, M.; Jannet Vennila, J.

2016-01-01

Two substituted aromatic carbonyl compounds (compounds 1 and 2) of 4-aminoantipyrine were synthesized by condensation of fluorine substituted benzoyl chlorides and 4-aminoantipyrine. The structures of synthesized derivatives were established on the basis of UV-Vis, IR, and Mass, 1H, 13C NMR and Fluorescence spectroscopy. Both compounds showed significant fluorescence emission and two broad emission bands were observed in the region at 340 nm and 450 nm on excitation at 280 nm. Theoretically to prove that the molecule has anticancer activity against cervical cancer cells, the compounds were analyzed for molecular docking interactions with HPV16-E7 target protein by Glide protocol. Furthermore, 4-aminoantipyrine derivatives were evaluated for their in vitro cytotoxic activity against human cervical cancer cells (SiHa) by MTT assay. Compound 1 showed two fold higher activity (IC50 = 0.912 μM) over compound 2, and its activity was similar to that of Pazopanib, suggesting that although the two compounds were chemically very similar the difference in substituent on the phenyl moiety caused changes in properties.

Metal-Enhanced Fluorescence from Nanoparticulate Zinc Films

PubMed Central

Aslan, Kadir; Previte, Michael J.R.; Zhang, Yongxia; Geddes, Chris D.

2009-01-01

A detailed study of metal-enhanced fluorescence (MEF) from fluorophores in the blue-to- red spectral region placed in close proximity to thermally evaporated zinc nanostructured films is reported. The zinc nanostructured films were deposited onto glass microscope slides as individual particles and were 1–10 nm in height and 20–100 nm in width, as characterized by Atomic Force Microscopy. The surface plasmon resonance peak of the zinc nanostructured films was ≈ 400 nm. Finite-difference time-domain calculations for single and multiple nanostructures organized in a staggered fashion on a solid support predict, as expected, that the electric fields are concentrated both around and between the nanostructures. Additionally, Mie scattering calculations show that the absorption and scattering components of the extinction spectrum are dominant in the UV and visible spectral ranges, respectively. Enhanced fluorescence emission accompanied by no significant changes in excited state lifetimes of fluorophores with emission wavelengths in the visible blue-to-red spectral range near-to zinc nanostructured films were observed, implying that MEF from zinc nanostructured films is mostly due to an electric field enhancement effect. PMID:19946356

Characterization of fluorescence in heat-treated silver-exchanged zeolites.

PubMed

De Cremer, Gert; Coutiño-Gonzalez, Eduardo; Roeffaers, Maarten B J; Moens, Bart; Ollevier, Jeroen; Van der Auweraer, Mark; Schoonheydt, Robert; Jacobs, Pierre A; De Schryver, Frans C; Hofkens, Johan; De Vos, Dirk E; Sels, Bert F; Vosch, Tom

2009-03-04

Thermal treatment of Ag(+)-exchanged zeolites yields discrete highly photostable luminescent clusters without formation of metallic nanoparticles. Different types of emitters with characteristic luminescence colors are observed, depending on the nature of the cocation, the amount of exchanged silver, and the host topology. The dominant emission bands in LTA samples are situated around 550 and 690 nm for the samples with, respectively, low and high silver content, while in FAU-type materials only a broad band around 550 nm is observed, regardless of the degree of exchange. Analysis of the fluorescent properties in combination with ESR spectroscopy suggests that a Ag(6)(+) cluster with doublet electronic ground state is associated with the appearance of the 690-nm emitter, having a decay of a few hundred microseconds. Tentatively, the nanosecond-decaying 550-nm emitter is assigned to the Ag(3)(+) cluster. This new class of photostable luminescent particles with tunable emission colors offers interesting perspectives for various applications such as biocompatible labels for intracellular imaging.

Engineering ESPT pathways based on structural analysis of LSSmKate red fluorescent proteins with large Stokes shift.

PubMed

Piatkevich, Kiryl D; Malashkevich, Vladimir N; Almo, Steven C; Verkhusha, Vladislav V

2010-08-11

LSSmKate1 and LSSmKate2 are monomeric red fluorescent proteins (RFPs) with large Stokes shifts (LSSs), which allows for efficient separation of absorbance and emission maxima, as well as for excitation with conventional two-photon laser sources. These LSSmKates differ by a single amino acid substitution at position 160 and exhibit absorbance maxima around 460 nm, corresponding to a neutral DsRed-like chromophore. However, excitation at 460 nm leads to fluorescence emission above 600 nm. Structures of LSSmKate1 and LSSmKate2, determined at resolutions of 2.0 and 1.5 A, respectively, revealed that the predominant DsRed-chromophore configurations are cis for LSSmKate1 but trans for LSSmKate2. Crystallographic and mutagenesis analyses, as well as isotope and temperature dependences, suggest that an excited-state proton transfer (ESPT) is responsible for the LSSs observed in LSSmKates. Hydrogen bonding between the chromophore hydroxyl and Glu160 in LSSmKate1 and a proton relay involving the chromophore tyrosine hydroxyl, Ser158, and the Asp160 carboxylate in LSSmKate2 represent the putative ESPT pathways. Comparisons with mKeima LSS RFP suggest that similar proton relays could be engineered in other FPs. Accordingly, we mutated positions 158 and 160 in several conventional red-shifted FPs, including mNeptune, mCherry, mStrawberry, mOrange, and mKO, and the resulting FP variants exhibited LSS fluorescence emission in a wide range of wavelengths from 560 to 640 nm. These data suggest that different chromophores formed by distinct tripeptides in different environments can be rationally modified to yield RFPs with novel photochemical properties.

[Chromophoric dissolved organic matter absorption characteristics with relation to fluorescence in typical macrophyte, algae lake zones of Lake Taihu].

PubMed

Zhang, Yun-lin; Qin, Bo-qiang; Ma, Rong-hua; Zhu, Guang-wei; Zhang, Lu; Chen, Wei-min

2005-03-01

Chromophoric dissolved organic matter (CDOM) represents one of the primary light-absorbing species in natural waters and plays a critical in determining the aquatic light field. CDOM shows a featureless absorption spectrum that increases exponentially with decreasing wavelength, which limits the penetration of biologically damaging UV-B radiation (wavelength from 280 to 320 nm) in the water column, thus shielding aquatic organisms. CDOM absorption measurements and their relationship with dissolved organic carbon (DOC), and fluorescence are presented in typical macrophyte and algae lake zone of Lake Taihu based on a field investigation in April in 2004 and lab analysis. Absorption spectral of CDOM was measured from 240 to 800 nm using a Shimadzu UV-2401PC UV-Vis recording spectrophotometer. Fluorescence with an excitation wavelength of 355 nm, an emission wavelength of 450 nm is measured using a Shimadzu 5301 spectrofluorometer. Concentrations of DOC ranged from 6.3 to 17.2 mg/L with an average of 9.08 +/- 2.66 mg/L. CDOM absorption coefficients at 280 nm and 355 nm were in the range of 11.2 - 32.6 m(-1) (average 17.46m(-1) +/- 5.75 m(-1) and 2.4 - 8.3 m(-1) (average 4.17m(-1) +/- 1.47 m(-l)), respectively. The values of the DOC-specific absorption coefficient at 355 nm ranged from 0.31 to 0.64 L x (mg x m)-1. Fluorescence emission at 450 nm, excited at 355 nm, had a mean value of 1.32nm(-1) +/- 0.84 nm(-1). A significant lake zone difference is found in DOC concentration, CDOM absorption coefficient and fluorescence, but not in DOC-specific absorption coefficient and spectral slope coefficient. This regional distribution pattern is in agreement with the location of sources of yellow substance: highest concentrations close to river mouth under the influence of river inflow, lower values in East Lake Taihu. The values of algae lake zone are obvious larger than those of macrophyte lake zone. In Meiliang Bay, CDOM absorption, DOC concentration and fluorescence tend to decreasing from inside to mouth of the Bay. The results show a good correlation between CDOM absorption and DOC coefficients during 280 - 500 nm short wavelength intervals. The R-square coefficient between CDOM absorption and DOC concentration decreases with the increase of wavelength from 280 to 500 nm. The significant linear regression correlations between fluorescence, DOC concentration and absorption coefficients were found at 355 nm. The exponential slope coefficients ranged from 13.0 to 16.4 microm(-1) with a mean value 14.37microm(-1) +/- 0.73microm(-1), 17.3microm(-1) - 20.3microm(-1) with a mean value 19.17microm(-1) +/- 0.84microm(-1) and 12.0microm(-1) - 15.8microm(-1) with a mean value 13.38microm(-1) +/- 0.82microm(-1) over the 280 - 500 nm, 280 - 360 nm and 360 - 440 nm intervals.

Fluorescence of size-expanded DNA bases: reporting on DNA sequence and structure with an unnatural genetic set.

PubMed

Krueger, Andrew T; Kool, Eric T

2008-03-26

We recently described the synthesis and helix assembly properties of expanded DNA (xDNA), which contains base pairs 2.4 A larger than natural DNA pairs. This designed genetic set is under study with the goals of mimicking the functions of the natural DNA-based genetic system and of developing useful research tools. Here, we study the fluorescence properties of the four expanded bases of xDNA (xA, xC, xG, xT) and evaluate how their emission varies with changes in oligomer length, composition, and hybridization. Experiments were carried out with short oligomers of xDNA nucleosides conjugated to a DNA oligonucleotide, and we investigated the effects of hybridizing these fluorescent oligomers to short complementary DNAs with varied bases opposite the xDNA bases. As monomer nucleosides, the xDNA bases absorb light in two bands: one at approximately 260 nm (similar to DNA) and one at longer wavelength ( approximately 330 nm). All are efficient violet-blue fluorophores with emission maxima at approximately 380-410 nm and quantum yields (Phifl) of 0.30-0.52. Short homo-oligomers of the xDNA bases (length 1-4 monomers) showed moderate self-quenching except xC, which showed enhancement of Phifl with increasing length. Interestingly, multimers of xA emitted at longer wavelengths (520 nm) as an apparent excimer. Hybridization of an oligonucleotide to the DNA adjacent to the xDNA bases (with the xDNA portion overhanging) resulted in no change in fluorescence. However, addition of one, two, or more DNA bases in these duplexes opposite the xDNA portion resulted in a number of significant fluorescence responses, including wavelength shifts, enhancements, or quenching. The strongest responses were the enhancement of (xG)n emission by hybridization of one or more adenines opposite them, and the quenching of (xT)n and (xC)n emission by guanines opposite. The data suggest multiple ways in which the xDNA bases, both alone and in oligomers, may be useful as tools in biophysical analysis and biotechnological applications.

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

Pletneva, Nadya V.; Pletnev, Vladimir Z.; Shemiakina, Irina I.

The wild type red fluorescent protein eqFP578 (from sea anemone Entacmaea quadricolor, {lambda}{sub ex} = 552 nm, {lambda}{sub em} = 578 nm) and its bright far-red fluorescent variant Katushka ({lambda}{sub ex} = 588 nm, {lambda}{sub em} = 635 nm) are characterized by the pronounced pH dependence of their fluorescence. The crystal structures of eqFP578f (eqFP578 with two point mutations improving the protein folding) and Katushka have been determined at the resolution ranging from 1.15 to 1.85 {angstrom} at two pH values, corresponding to low and high level of fluorescence. The observed extinguishing of fluorescence upon reducing pH in eqFP578f andmore » Katushka has been shown to be accompanied by the opposite trans-cis and cis-trans chromophore isomerization, respectively. Asn143, Ser158, His197 and Ser143, Leu174, and Arg197 have been shown to stabilize the respective trans and cis fluorescent states of the chromophores in eqFP578f and Katushka at higher pH. The cis state has been suggested as being primarily responsible for the observed far-red shift of the emission maximum of Katushka relative to that of eqFP578f.« less

Analogues of uracil nucleosides with intrinsic fluorescence (NIF-analogues): synthesis and photophysical properties.

PubMed

Segal, Meirav; Fischer, Bilha

2012-02-28

Uridine cannot be utilized as fluorescent probe due to its extremely low quantum yield. For improving the uracil fluorescence characteristics we extended the natural chromophore at the C5 position by coupling substituted aromatic rings directly or via an alkenyl or alkynyl linker to create fluorophores. Extension of the uracil base was achieved by treating 5-I-uridine with the appropriate boronic acid under the Suzuki coupling conditions. Analogues containing an alkynyl linker were obtained from 5-I-uridine and the suitable boronic acid in a Sonogashira coupling reaction. The uracil fluorescent analogues proposed here were designed to satisfy the following requirements: a minimal chemical modification at a position not involved in base-pairing, resulting in relatively long absorption and emission wavelengths and high quantum yield. 5-((4-Methoxy-phenyl)-trans-vinyl)-2'-deoxy-uridine, 6b, was found to be a promising fluorescent probe. Probe 6b exhibits a quantum yield that is 3000-fold larger than that of the natural chromophore (Φ 0.12), maximum emission (478 nm) which is 170 nm red shifted as compared to uridine, and a Stokes shift of 143 nm. In addition, since probe 6b adopts the anti conformation and S sugar puckering favored by B-DNA, it makes a promising nucleoside analogue to be incorporated in an oligonucleotide probe for detection of genetic material.

Early quantitative method for measuring germination in non-green spores of Dryopteris paleacea using an epifluorescence-microscope technique

NASA Technical Reports Server (NTRS)

Scheuerlein, R.; Wayne, R.; Roux, S. J.

1988-01-01

A method is described to determine germination by blue-light excited red fluorescence in the positively photoblastic spores of Dryopteris paleacea Sw. This fluorescence is due to chlorophyll as evidenced from 1) a fluorescence-emission spectrum in vivo, where a bright fluorescence around 675 nm is obtained only in red light (R)-irradiated spores and 2) in vitro measurements with acetone extracts prepared from homogenized spores. Significant amounts of chlorophyll can be found only in R-treated spores; this chlorophyll exhibits an emission band around 668 nm, when irradiated with 430 nm light at 21 degrees C. Compared to other criteria for germination, such as swelling of the cell, coat splitting, greening, and rhizoid formation, which require longer periods after induction for their expression, chlorophyll fluorescence can be used to quantify germination after two days. This result is confirmed by fluence-response curves for R-induced spore germination; the same relationship between applied R and germination is obtained by the evaluation with the epifluorescence method 2 days after the light treatment as compared with the evaluation with bright-field microscopy 5 days after the inducing R. Using this technique we show for the first time that Ca2+ contributes to the signal-transduction chain in phytochrome-mediated chlorophyll synthesis in spores of Dryopteris paleacea.

Optically-Enhanced, Near-IR, Silver Cluster Emission Altered by Single Base Changes in the DNA Template

PubMed Central

Fan, Chaoyang; Story, Sandra P.; Sengupta, Bidisha; Sartin, Matthew; Hsiang, Jung-Cheng; Perry, Joseph W.

2011-01-01

Few-atom silver clusters harbored by DNA are promising fluorophores due to their high molecular brightness along with their long- and short-term photostability. Furthermore, their emission rate can be enhanced when co-illuminated with low-energy light that optically depopulates the fluorescence-limiting dark state. The photophysical basis for this effect is evaluated for two near infrared-emitting clusters. Clusters emitting at ~800 nm form with C3AC3AC3TC3A and C3AC3AC3GC3A and both exhibit a trap state with λmax ~ 840 nm and an absorption cross section of 5–6 × 10−16 cm2/molec that can be optically depopulated. Transient absorption spectra, complemented by fluorescence correlation spectroscopy studies, show that the dark state has an inherent lifetime of 3–4 μs and that absorption from this state is accompanied by photoinduced crossover back to the emissive manifold of states with an action cross section of ~2 × 10−18 cm2/molec. Relative to C3AC3AC3TC3A, C3AC3AC3GC3A produces a longer-lived trap state and permits more facile passage back to the emissive manifold. With the C3AC3AC3AC3G template, a spectrally distinct cluster forms having emission at ~900 nm and its trap state has a ~four-fold shorter lifetime. These studies of optically-gated fluorescence bolster the critical role of the nucleobases on both the formation and excited state dynamics of these highly emissive metallic clusters. PMID:21568292

Near-Infrared Fluorescence Imaging Guided Therapy: Molecular Beacon-Based Photosensitizers Triggered by Breast Cancer-Specific mRNA

DTIC Science & Technology

2007-05-01

1989) Specific targeting and toxicity of sulphonated aluminium phthalocyanine photosensitised liposomes directed to cells by monoclonal antibody in...fluorescence emission wavelengths (λabs 825nm; λem 840nm)[1]. Since the native BChl is very unstable and undergoes rapid oxidation to the chlorin state...noticeable dark toxicity in experimental condition, the viability of cells incubated with 2μM of P30C, P30 or r- P30C without light being the same as

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

NASA Astrophysics Data System (ADS)

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.

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

PubMed

Cervantes-Martínez, Jesús; Flores-Hernández, Ricardo; Rodríguez-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.

Disposal of Energy by UV-B Sunscreens

NASA Astrophysics Data System (ADS)

Nordlund, Thomas; Krishnan, Rajagopal

2008-03-01

Ideal sunscreens absorb dangerous UV light and dispose of the energy safely. ``Safe disposal'' usually means conversion to heat. However, efficient absorption entails a high radiative rate, which implies high energy-transfer and other rates, unless some process intervenes to ``defuse'' the excited state. We studied the excited-state kinetics of three UV-B (290-320 nm) sunscreens by absorption, steady-state and time-resolved fluorescence. Excited-state rate analysis suggests that some sunscreens have low radiative-rate ``dark'' states, in addition to normal excited states.* We deduce dark states when sunscreens of high extinction coefficient do not show lifetimes and total emission consistent with such high radiative rates. A high radiative rate, accompanied by efficient fluorescence emission and/or transfer, may be unfavorable for a sunscreen. In spite of its dark excited state, padimate O shows significant re-emission of light in the UV-A (320-400 nm) and energy transfer to a natural component of excised skin, probably collagen. * Krishnan, R. and T.M. Nordlund (2007) J. Fluoresc. DOI 10.1007/s10895-007-0264-3.

[Spectral properties of new photosensitizers for photodynamic diagnosis and therapy].

PubMed

Li, Bu-hong; Xie, Shu-sen; Lu, Zu-kang

2002-12-01

The spectral properties of new photosensitizer ZnPcS2P2, PsD-007 and HMME, as well as traditional photosensitizer HpD have been studied by comparing their spectra in physiological saline and in physiological saline with 10 percent serum. Experimental results show that the maximum absorption peaks for PsD-007, HMME and HpD in the physiological saline with 10 percent serum appear at 400 nm in the soret region, while at 670 nm for ZnPcS2P2. The fluorescence excitation spectra closely resemble the absorption spectra. When excited by the light at the wavelengths of 413 and 514.5 nm, the fluorescence emission peaks for PsD-007, HMME and HpD appear at 625 and 690 nm, respectively. The fluorescent excitation efficiency of the same photosensitizer with the same concentration excited by the light at the wavelength of 413 nm is about three fold higher than that at 514.5 nm. Furthermore, the fluorescent excitation efficiency is the highest for HMME, but is lower for HpD and lowest for PsD-007. These results are significant in the selection of photosensitizers for photodynamic diagnosis and therapy.

Plant abiotic stress diagnostic by laser induced chlorophyll fluorescence spectral analysis of in vivo leaf tissue of biofuel species

NASA Astrophysics Data System (ADS)

Gouveia-Neto, Artur S.; Silva, Elias A., Jr.; Costa, Ernande B.; Bueno, Luciano A.; Silva, Luciana M. H.; Granja, Manuela M. C.; Medeiros, Maria J. L.; Câmara, Terezinha J. R.; Willadino, Lilia G.

2010-02-01

Laser induced fluorescence is exploited to evaluate the effect of abiotic stresses upon the evolution and characteristics of in vivo chlorophyll emission spectra of leaves tissues of brazilian biofuel plants species(Saccharum officinarum and Jatropha curcas). The chlorophyll fluorescence spectra of 20 min predarkened intact leaves were studied employing several excitation wavelengths in the UV-VIS spectral region. Red(Fr) and far-red (FFr) chlorophyll fluorescence emission signals around 685 nm and 735 nm, respectively, were analyzed as a function of the stress intensity and the time of illumination(Kautsky effect). The Chl fluorescence ratio Fr/FFr which is a valuable nondestructive indicator of the chlorophyll content of leaves was investigated during a period of time of 30 days. The dependence of the Chl fluorescence ratio Fr/FFr upon the intensity of the abiotic stress(salinity) was examined. The results indicated that the salinity plays a major hole in the chlorophyll concentration of leaves in both plants spieces, with a significant reduction in the chlorophyll content for NaCl concentrations in the 25 - 200 mM range. The laser induced chlorophyll fluorescence analysis allowed detection of damage caused by salinity in the early stages of the plants growing process, and can be used as an early-warning indicator of salinity stress

Effects of gold nanoparticles on the photophysical and photosynthetic parameters of leaves and chloroplasts.

PubMed

Torres, Rocio; Diz, Virginia E; Lagorio, M Gabriela

2018-04-18

Effects of gold nanoparticles (average diameter: 10-14 nm) on leaves and chloroplasts have been studied. Gold nanoparticles (AuNPs) quenched significantly chlorophyll fluorescence when introduced both in intact leaves and isolated chloroplasts. Additionally, the fluorescence spectra corrected for light re-absorption processes showed a net decrease in the fluorescence ratio calculated as the quotient between the maximum fluorescence at 680 and 735 nm. This fact gave evidence for a reduction in the fluorescence emission of the PSII relative to that of the PSI. Strikingly, the photosynthetic parameters derived from the analysis of the slow phase of Kautsky's kinetics, the rate of oxygen evolution and the rate of photo-reduction of 2,6-dichlorophenolindophenol were increased in the presence of AuNPs indicating an apparent greater photosynthetic capacity. The observed results were consistent with an electron transfer process from the excited PSII, which was thermodynamically possible, and which competed with both the electron transport process that initiated photosynthesis and the deactivation of the excited PSII by fluorescence emission. Additionally, it is here explained, in terms of a completely rational kinetic scheme and their corresponding algebraic expressions, why the photosynthetic parameters and the variable and non-variable fluorescence of chlorophyll are modified in a photosynthetic tissue containing gold nanoparticles.

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

PubMed

Žoldák, Gabriel; Jancura, Daniel; Sedlák, Erik

2017-06-01

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

Saccharide sensing molecules having enhanced fluorescent properties

DOEpatents

Satcher Jr., Joe H.; Lane, Stephen M.; Darrow, Christopher B.; Cary, Douglas R.; Tran, Joe Anh

2004-01-06

The present invention provides formulae for fluorescent compounds that have a number of properties which make them uniquely suited for use in sensors of analytes such as saccharides. The advantageous fluorescent properties include favorable excitation wavelengths, emission wavelengths, fluorescence lifetimes, and photostability. Additional advantageous properties include enhanced aqueous solubility, as well as temperature and pH sensitivity. The compound comprises an aryl or a substituted phenyl botonic acid that acts as a substrate recognition component, a fluorescence switch component, and a fluorophore. Fluorescent compounds are described that are excited at wavelengths greater than 400 nm and emit at wavelengths greater than 450 nm, which is advantageous for optical transmission through skin. The fluorophore is typically selected from transition metal-ligand complexes and thiazine, oxazine, oxazone, or oxazine-one as well as anthracene compounds. The fluorescent compound can be immobilized in a glucose permeable biocompatible polymer matrix that is implantable below the skin.

Multispectral guided fluorescence diffuse optical tomography using upconverting nanoparticles

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

Svenmarker, Pontus, E-mail: pontus.svenmarker@physics.umu.se; Department of Physics, Umeå University, SE-901 87 Umeå; Centre for Microbial Research

2014-02-17

We report on improved image detectability for fluorescence diffuse optical tomography using upconverting nanoparticles doped with rare-earth elements. Core-shell NaYF{sub 4}:Yb{sup 3+}/Er{sup 3+}@NaYF{sub 4} upconverting nanoparticles were synthesized through a stoichiometric method. The Yb{sup 3+}/Er{sup 3+} sensitizer-activator pair yielded two anti-Stokes shifted fluorescence emission bands at 540 nm and 660 nm, here used to a priori estimate the fluorescence source depth with sub-millimeter precision. A spatially varying regularization incorporated the a priori fluorescence source depth estimation into the tomography reconstruction scheme. Tissue phantom experiments showed both an improved resolution and contrast in the reconstructed images as compared to not using any amore » priori information.« less

Field trial of a dual-wavelength fluorescent emission (L.I.F.E.) instrument and the Magma White rover during the MARS2013 Mars analog mission.

PubMed

Groemer, Gernot; Sattler, Birgit; Weisleitner, Klemens; Hunger, Lars; Kohstall, Christoph; Frisch, Albert; Józefowicz, Mateusz; Meszyński, Sebastian; Storrie-Lombardi, Michael; Bothe, Claudia; Boyd, Andrea; Dinkelaker, Aline; Dissertori, Markus; Fasching, David; Fischer, Monika; Föger, Daniel; Foresta, Luca; Frischauf, Norbert; Fritsch, Lukas; Fuchs, Harald; Gautsch, Christoph; Gerard, Stephan; Goetzloff, Linda; Gołebiowska, Izabella; Gorur, Paavan; Groemer, Gerhard; Groll, Petra; Haider, Christian; Haider, Olivia; Hauth, Eva; Hauth, Stefan; Hettrich, Sebastian; Jais, Wolfgang; Jones, Natalie; Taj-Eddine, Kamal; Karl, Alexander; Kauerhoff, Tilo; Khan, Muhammad Shadab; Kjeldsen, Andreas; Klauck, Jan; Losiak, Anna; Luger, Markus; Luger, Thomas; Luger, Ulrich; McArthur, Jane; Moser, Linda; Neuner, Julia; Orgel, Csilla; Ori, Gian Gabriele; Paternesi, Roberta; Peschier, Jarno; Pfeil, Isabella; Prock, Silvia; Radinger, Josef; Ragonig, Christoph; Ramirez, Barbara; Ramo, Wissam; Rampey, Mike; Sams, Arnold; Sams, Elisabeth; Sams, Sebastian; Sandu, Oana; Sans, Alejandra; Sansone, Petra; Scheer, Daniela; Schildhammer, Daniel; Scornet, Quentin; Sejkora, Nina; Soucek, Alexander; Stadler, Andrea; Stummer, Florian; Stumptner, Willibald; Taraba, Michael; Tlustos, Reinhard; Toferer, Ernst; Turetschek, Thomas; Winter, Egon; Zanella-Kux, Katja

2014-05-01

Abstract We have developed a portable dual-wavelength laser fluorescence spectrometer as part of a multi-instrument optical probe to characterize mineral, organic, and microbial species in extreme environments. Operating at 405 and 532 nm, the instrument was originally designed for use by human explorers to produce a laser-induced fluorescence emission (L.I.F.E.) spectral database of the mineral and organic molecules found in the microbial communities of Earth's cryosphere. Recently, our team had the opportunity to explore the strengths and limitations of the instrument when it was deployed on a remote-controlled Mars analog rover. In February 2013, the instrument was deployed on board the Magma White rover platform during the MARS2013 Mars analog field mission in the Kess Kess formation near Erfoud, Morocco. During these tests, we followed tele-science work flows pertinent to Mars surface missions in a simulated spaceflight environment. We report on the L.I.F.E. instrument setup, data processing, and performance during field trials. A pilot postmission laboratory analysis determined that rock samples acquired during the field mission exhibited a fluorescence signal from the Sun-exposed side characteristic of chlorophyll a following excitation at 405 nm. A weak fluorescence response to excitation at 532 nm may have originated from another microbial photosynthetic pigment, phycoerythrin, but final assignment awaits development of a comprehensive database of mineral and organic fluorescence spectra. No chlorophyll fluorescence signal was detected from the shaded underside of the samples.

Photoabsorption and photodissociation of molecules important in the interstellar medium

NASA Technical Reports Server (NTRS)

Lee, Long C.; Suto, Masako

1991-01-01

The photoabsorption, photodissociation, and fluorescence cross sections of interstellar molecules are measured at 90 to 250 nm. These quantitative optical data are needed for the understanding of the formation and destruction processes of molecules under the intense interstellar UV radiation field. Research covering the following topics is presented: (1) fluorescences from photoexcitation of CH4, CH3OH, and CH3SH; (2) NO gamma emission from photoexcitation of NO; (3) photoexcitation cross sections of aromatic molecules; (4) IR emission from UV excitation of HONO2; (5) IR emission from UV excitation of benzene and methyl-derivitives; and (6) IR emission from UV excitation of polycyclic aromatic hydrocarbon molecules.

Red-shifted aequorin-based bioluminescent reporters for in vivo imaging of Ca2 signaling.

PubMed

Curie, Thomas; Rogers, Kelly L; Colasante, Cesare; Brûlet, Philippe

2007-01-01

Real-time visualization of calcium (Ca(2+)) dynamics in the whole animal will enable important advances in understanding the complexities of cellular function. The genetically encoded bioluminescent Ca(2+) reporter green fluorescent protein-aequorin (GA) allows noninvasive detection of intracellular Ca(2+) signaling in freely moving mice. However, the emission spectrum of GA is not optimal for detection of activity from deep tissues in the whole animal. To overcome this limitation, two new reporter genes were constructed by fusing the yellow fluorescent protein (Venus) and the monomeric red fluorescent protein (mRFP1) to aequorin. Transfer of aequorin chemiluminescence energy to Venus (VA) is highly efficient and produces a 58 nm red shift in the peak emission spectrum of aequorin. This substantially improves photon transmission through tissue, such as the skin and thoracic cage. Although the Ca(2+)-induced bioluminescence spectrum of mRFP1-aequorin (RA) is similar to that of aequorin, there is also a small peak above 600 nm corresponding to the peak emission of mRFP1. Small amounts of energy transfer between aequorin and mRFP1 yield an emission spectrum with the highest percentage of total light above 600 nm compared with GA and VA. Accordingly, RA is also detected with higher sensitivity from brain areas. VA and RA will therefore improve optical access to Ca(2+) signaling events in deeper tissues, such as the heart and brain, and offer insight for engineering new hybrid molecules.

The role of aromatic side chain residues in micelle binding by pancreatic colipase. Fluorescence studies of the porcine and equine proteins.

PubMed Central

McIntyre, J C; Hundley, P; Behnke, W D

1987-01-01

Fluorescence techniques have been employed to study the interaction of porcine and equine colipase with pure taurodeoxycholate and mixed micelles. Nitrotyrosine-55 of porcine colipase is obtained by modification with tetranitromethane (low excess, in the presence of taurodeoxycholate) of the protein followed by gel filtration and ion-exchange chromatography. Verification of the residue modified was obtained by h.p.l.c. peptide purification and sequence analysis. Reduction and quantitative reaction with dansyl chloride yields a fluorescent derivative that is twice as active in conjunction with lipase as is native colipase and that exhibits a strong emission band at 550 nm. Addition of micellar concentrations of taurodeoxycholate causes a 4.3-fold increase in the emission maximum as well as a 70 nm blue shift to 480 nm. Inclusion of oleic acid to form a mixed micelle reduces these spectral effects. Scatchard analysis of the data yield a Kd of 6.8 X 10(-4) M and a single colipase-binding site for taurodeoxycholate micelles. The data, by analogy to a phospholipase system, are consistent with a direct insertion of dansyl-NH-tyrosine-55 into the micelle. The presence of a single tryptophan residue (Trp-52) in equine colipase provides an intrinsic fluorescent probe for studying protein-micelle interaction. The emission maximum of horse colipase at 345 nm indicates a solvent-accessible tryptophan residue which becomes less so on binding of micelles. A blue shift of 8 nm and a 2-fold increase in amplitude is indicative of a more hydrophobic environment for tryptophan induced by taurodeoxycholate micelles. There is also a decrease in KSV for acrylamide quenching in the presence of micelles, which further supports a loss of solvent accessibility. The most dramatic pH effects are observed with KI quenching, and may indicate the presence of negative charges near Trp-52. PMID:3663193

A novel approach to dual excitation ratiometric optical mapping of cardiac action potentials with di-4-ANEPPS using pulsed LED excitation.

PubMed

Bachtel, Andrew D; Gray, Richard A; Stohlman, Jayna M; Bourgeois, Elliot B; Pollard, Andrew E; Rogers, Jack M

2011-07-01

We developed a new method for ratiometric optical mapping of transmembrane potential (V(m)) in cardiac preparations stained with di-4-ANEPPS. V(m)-dependent shifts of excitation and emission spectra establish two excitation bands (<481 and >481 nm) that produce fluorescence changes of opposite polarity within a single emission band (575-620 nm). The ratio of these positive and negative fluorescence signals (excitation ratiometry) increases V(m) sensitivity and removes artifacts common to both signals. We pulsed blue (450 ± 10 nm) and cyan (505 ± 15 nm) light emitting diodes (LEDs) at 375 Hz in alternating phase synchronized to a camera (750 frames-per-second). Fluorescence was bandpass filtered (585 ± 20 nm). This produced signals with upright (blue) and inverted (cyan) action potentials (APs) interleaved in sequential frames. In four whole swine hearts with motion chemically arrested, fractional fluorescence for blue, cyan, and ratio signals was 1.2 ± 0.3%, 1.2 ± 0.3%, and 2.4 ± 0.6%, respectively. Signal-to-noise ratios were 4.3 ± 1.4, 4.0 ± 1.2, and 5.8 ± 1.9, respectively. After washing out the electromechanical uncoupling agent, we characterized motion artifact by cross-correlating blue, cyan, and ratio signals with a signal with normal AP morphology. Ratiometry improved cross-correlation coefficients from 0.50 ± 0.48 to 0.81 ± 0.25, but did not cancel all motion artifacts. These findings demonstrate the feasibility of pulsed LED excitation ratiometry in myocardium. © 2011 IEEE

A Novel Approach to Dual Excitation Ratiometric Optical Mapping of Cardiac Action Potentials with Di-4-ANEPPS using Pulsed LED Excitation

PubMed Central

Bachtel, Andrew D.; Gray, Richard A.; Stohlman, Jayna M.; Bourgeois, Elliot B.; Pollard, Andrew E.

2011-01-01

We developed a new method for ratiometric optical mapping of transmembrane potential (Vm) in cardiac preparations stained with di-4-ANEPPS. Vm-dependent shifts of excitation and emission spectra establish two excitation bands (<481 and >481 nm) that produce fluorescence changes of opposite polarity within a single emission band (575–620 nm). The ratio of these positive and negative fluorescence signals (excitation ratiometry) increases Vm sensitivity and removes artifacts common to both signals. We pulsed blue (450±10 nm) and cyan (505±15 nm) light emitting diodes (LEDs) at 375 Hz in alternating phase synchronized to a camera (750 frames-per-second). Fluorescence was bandpass filtered (585±20 nm). This produced signals with upright (blue) and inverted (cyan) action potentials (APs) interleaved in sequential frames. In 4 whole swine hearts with motion chemically arrested, fractional fluorescence for blue, cyan, and ratio signals was 1.2±0.3%, 1.2±0.3%, and 2.4±0.6%, respectively. Signal-to-noise ratios were 4.3±1.4, 4.0±1.2, and 5.8±1.9, respectively. After washing out the electromechanical uncoupling agent, we characterized motion artifact by cross-correlating blue, cyan, and ratio signals with a signal with normal AP morphology. Ratiometry improved cross-correlation coefficients from 0.50±0.48 to 0.81±0.25, but did not cancel all motion artifacts. These findings demonstrate the feasibility of pulsed LED excitation ratiometry in myocardium. PMID:21536528

Light-induced flickering of DsRed provides evidence for distinct and interconvertible fluorescent states.

PubMed Central

Malvezzi-Campeggi, F; Jahnz, M; Heinze, K G; Dittrich, P; Schwille, P

2001-01-01

Green fluorescent protein (GFP) from jellyfish Aequorea victoria, the powerful genetically encoded tag presently available in a variety of mutants featuring blue to yellow emission, has found a red-emitting counterpart. The recently cloned red fluorescent protein DsRed, isolated from Discosoma corals (), with its emission maximum at 583 nm, appears to be the long awaited tool for multi-color applications in fluorescence-based biological research. Studying the emission dynamics of DsRed by fluorescence correlation spectroscopy (FCS), it can be verified that this protein exhibits strong light-dependent flickering similar to what is observed in several yellow-shifted mutants of GFP. FCS data recorded at different intensities and excitation wavelengths suggest that DsRed appears under equilibrated conditions in at minimum three interconvertible states, apparently fluorescent with different excitation and emission properties. Light absorption induces transitions and/or cycling between these states on time scales of several tens to several hundreds of microseconds, dependent on excitation intensity. With increasing intensity, the emission maximum of the static fluorescence continuously shifts to the red, implying that at least one state emitting at longer wavelength is preferably populated at higher light levels. In close resemblance to GFP, this light-induced dynamic behavior implies that the chromophore is subject to conformational rearrangements upon population of the excited state. PMID:11509387

Light-induced fluorescence changes in Phycomyces: evidence for blue light-receptor associated flavo-semiquinones.

PubMed

Galland, Paul; Tölle, Nadja

2003-10-01

Light-induced fluorescence changes (LIFCs) were detected in sporangiophores of the blue-light-sensitive fungus Phycomyces blakesleeanus (Burgeff). The LIFCs can be utilized as a spectrophotometric assay for blue-light photoreceptors and for the in vivo characterization of their photochemical primary reactions. Blue-light irradiation of sporangiophores elicited a transient decrease and subsequent regeneration of flavin-like fluorescence emission at 525 nm. The signals recovered in darkness in about 120 min. In contrast to blue light, near-UV (370 nm) caused an increase in the fluorescence emission at 525 nm. Because the LIFCs were altered in a light-insensitive madC mutant with a defective photoreceptor, the fluorescence changes must be associated with early photochemical events of the transduction chain. Action spectra for the fluorescence changes at 525 nm showed major peaks near 470 and 600 nm. Double-pulse experiments involving two consecutive pulses of either blue and near-UV, blue and red, or near-UV and red showed that the responses depended on the sequence in which the different wavelengths were applied. The results indicate a blue-light receptor with intermediates in the near-UV, blue and red spectral regions. We explain the results in the framework of a general model, in which the three redox states of the flavin photoreceptor, the oxidized flavin (Fl), the flavo-semiquinone (FlH*), and the flavo-hydroquinone (FlH2) are each acting as chromophores with their own characteristic photochemical primary reactions. These consist of the photoreduction of the oxidized flavin generating semiquinone, the photoreduction of the semiquinone generating hydroquinone, and the photooxidation of the flavo-hydroquinone regenerating the pool of oxidized flavins. The proposed mechanism represents a photocycle in which two antagonistic photoreceptor forms, Fl and FlH2, determine the pool size of the biological effector molecule, the flavo-semiquinone. The redox changes that are associated with the photocycle are maintained by redox partners, pterins, that function in the near-UV as secondary chromophores.

Activation energy of light induced isomerization of resveratrol.

PubMed

Figueiras, Teresa Sofia; Neves-Petersen, Maria Teresa; Petersen, Steffen B

2011-09-01

Isomerization of trans-stilbenes is known to be induced by light. The two isomers have distinct absorption, fluorescence excitation and emission spectra. Resveratrol, 3,4',5-trihydroxystilbene, is a member of the stilbene family. The interest of the scientific community in resveratrol has increased over the last years due to its biomedical properties. Whereas there is a growing confidence that trans-resveratrol is non-toxic, very little is known about the pharmacology of cis-resveratrol. Of this very reason there is considerable interest in knowing the energetics of the trans-cis conversion. Cis-resveratrol is characterized by a large fluorescence quantum yield when compared to trans-resveratrol. In the present paper we report a detailed analysis of the spectral changes induced in trans-resveratrol upon 260 nm excitation for different time periods. Spectral changes have been monitored with UV-visible absorption and steady-state fluorescence spectroscopy at pH 4 at 20, 25, 30, 35, 40, 45 and 50 °C. Continuous 260 nm excitation induces a blue shift in the absorption and fluorescence excitation spectra of resveratrol and a 14 nm blue shift in its fluorescence emission. The photoisomerization yield is reported as a function of 260 nm excitation time. 330 min continuous excitation led to ~60% isomerization yield. The kinetics of trans-cis isomerization has been monitored following the increase in fluorescence quantum yield upon continuous 260 nm excitation of trans-resveratrol. The study was carried out at the above mentioned temperatures in order to obtain the Arrhenius activation energy of photoisomerization. Activation energy and pre-exponential factor were 3.7 ± 0.3 kcal.mol(-1) and 10.6 ± 1.6 s(-1), respectively. The activation energy is comparable with previously reported values for the photoisomerization of other stilbenes.

Spectroscopic studies, fluorescence quenching by molecular oxygen and amplified spontaneous emission of 1,4-bis [2-(2-pyridyl) vinyl] benzene (P2VB) diolefinic laser dye

NASA Astrophysics Data System (ADS)

El-Daly, Samy A.; Ebeid, E. M.

2014-04-01

The UV-visible electronic absorption spectra, molar absorptivity, fluorescence spectra, fluorescence quantum yield and excited state lifetime of 1,4-bis [2-(2-pyridyl) vinyl] benzene P2VB were measured in different solvents. The fluorescence quenching of P2VB by molecular oxygen was also studied using lifetime measurements. A 2 × 10-4 mol dm-3 solution of P2VB in dimethyl formamide (DMF) gave amplified spontaneous emission (ASE) in blue spectral region with emission maximum at 420 nm upon pumping by 337.1 nitrogen laser pulse. The photochemical quantum yields (ϕc) of trans-cis photoisomerization of P2VB were calculated in different organic solvents. The photoreactivity of P2VB are also studied PMMA matrix.

Two-photon pumped amplified spontaneous emission based on all-inorganic perovskite nanocrystals embedded with gold nanorods

NASA Astrophysics Data System (ADS)

Liu, Shaoying; Fang, Xiaohui; Wang, Yimeng; Zhang, Xinping

2018-07-01

CsPbBr3 nanocrystals have attracted great interest owing to their high fluorescence quantum efficiency, adjustable photoluminescence wavelength, and good stability. We report a device that consists of disordered gold nanorods underneath a film of CsPbBr3 nanocrystals. Two-photon pumping using femtosecond laser pulses at 800 nm enables amplified spontaneous emission (ASE) at about 523 nm. In this work, a narrow peak with linewidth of 5 nm is observed when the pump fluence reaches a low threshold of 0.65 mJ/cm2. The results show that plasmonic resonance of gold nanorods improves the emission transition rate and enables low threshold ASE.

Controlling cross pumping between C-N and C-H vibration in nitromethane by selective fluorescence-enhanced stimulated Raman scattering.

PubMed

Wang, Shenghan; Fang, Wenhui; Li, Tianyu; Li, Fangfang; Sun, Chenglin; Li, Zuowei; Men, Zhiwei

2016-05-02

To investigate the vibrational features of nitromethane (NM), which is a kind of energy material and a well known low-sensitivity and high explosive, experiments are performed to obtain the stimulated Raman scattering (SRS) of NM by employing a 532 nm pulsed pump laser. The Raman signal involves two stimulated emissions at 918 and 2,963 cm^-1, attributed to the C-N and C-H stretching vibrations, respectively. To overcome the complexity of cross pump in the pure NM, one stimulated Raman radiation is chosen as a pump source to excite the other Raman mode. Two fluorescence dyes were added to selectively enhance each Raman cross section. By internally seeding the Raman gain medium with fluorescent photons, a significant modification in the stimulated Raman scattering spectrum has been observed. The enhanced Stokes emission at 918 cm^-1 was able to induce the 2,963 cm^-1 vibration mode when the all-trans-β-carotene was internal seeding in the NM, while the Raman radiation at 2,963 cm^-1 was enhanced to excite the C-N mode with the addition of m-Cresol purple. The output energy of both 918 and 2,963 cm^-1 under different input energy was also measured to illustrate this result.

Yellow fluorescent protein phiYFPv (Phialidium): structure and structure-based mutagenesis

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

Pletneva, Nadya V.; Pletnev, Vladimir Z., E-mail: vzpletnev@gmail.com; Souslova, Ekaterina

The yellow fluorescent protein phiYFPv with improved folding has been developed from the spectrally identical wild-type phiYFP found in the marine jellyfish Phialidium. The yellow fluorescent protein phiYFPv (λ{sub em}{sup max} ≃ 537 nm) with improved folding has been developed from the spectrally identical wild-type phiYFP found in the marine jellyfish Phialidium. The latter fluorescent protein is one of only two known cases of naturally occurring proteins that exhibit emission spectra in the yellow–orange range (535–555 nm). Here, the crystal structure of phiYFPv has been determined at 2.05 Å resolution. The ‘yellow’ chromophore formed from the sequence triad Thr65-Tyr66-Gly67 adoptsmore » the bicyclic structure typical of fluorophores emitting in the green spectral range. It was demonstrated that perfect antiparallel π-stacking of chromophore Tyr66 and the proximal Tyr203, as well as Val205, facing the chromophore phenolic ring are chiefly responsible for the observed yellow emission of phiYFPv at 537 nm. Structure-based site-directed mutagenesis has been used to identify the key functional residues in the chromophore environment. The obtained results have been utilized to improve the properties of phiYFPv and its homologous monomeric biomarker tagYFP.« less

Seasonal characterization of CDOM for lakes in semi-arid regions of Northeast China using excitation-emission matrices fluorescence and parallel factor analysis (EEM-PARAFAC)

NASA Astrophysics Data System (ADS)

Zhao, Y.; Song, K.; Wen, Z.; Li, L.; Zang, S.; Shao, T.; Li, S.; Du, J.

2015-04-01

The seasonal characteristics of fluorescence components in CDOM for lakes in the semi-arid region of Northeast China were examined by excitation-emission matrices fluorescence and parallel factor analysis (EEM-PARAFAC). Two humic-like peaks C1 (Ex/Em = 230, 300/425 nm) and C2 (Ex/Em = 255, 350/460 nm) and two protein-like B (Ex/Em = 220, 275/320 nm) and T (Ex/Em = 225, 290/360 nm) peaks were identified using PARAFAC. The average fluorescence intensity of the four components differed with seasonal variation from June and August 2013 to February and April 2014. The total fluorescence intensity significantly varied from 2.54 ± 0.68 nm-1 in June to the mean value 1.93 ± 0.70 nm-1 in August 2013, and then increased to 2.34 ± 0.92 nm-1 in February and reduced to the lowest 1.57 ± 0.55 nm-1 in April 2014. In general, the fluorescence intensity was dominated by peak C1, indicating that most part of CDOM for inland waters being investigated in this study was originated from phytoplankton degradation. The lowest C2 represents only a small portion of CDOM from terrestrial imported organic matter to water bodies through rainwash and soil leaching. The two protein-like intensities (B and T) formed in situ through microbial activity have almost the same intensity. Especially, in August 2013 and February 2014, the two protein-like peaks showed obviously difference from other seasons and the highest C1 (1.02 nm-1) was present in February 2014. Components 1 and 2 exhibited strong linear correlation (R2 = 0.633). There were significantly positive linear relationships between CDOM absorption coefficients a(254) (R2 = 0.72, 0.46, p < 0.01), a(280) (R2 = 0.77, 0.47, p < 0.01), a(350) (R2 = 0.76, 0.78, p < 0.01) and Fmax for two humic-like components (C1 and C2), respectively. A close relationship (R2 = 0.931) was found between salinity and DOC. However, almost no obvious correlation was found between salinity and EEM-PARAFAC extracted components except for C3 (R2 = 0.469). Results from this investigation demonstrate that the EEM-PARAFAC technique can be used to evaluate the seasonal dynamics of CDOM fluorescence components for inland waters in semi-arid regions of Northeast China.

Color-tunable mixed photoluminescence emission from Alq3 organic layer in metal-Alq3-metal surface plasmon structure.

PubMed

Chen, Nai-Chuan; Liao, Chung-Chi; Chen, Cheng-Chang; Fan, Wan-Ting; Wu, Jin-Han; Li, Jung-Yu; Chen, Shih-Pu; Huang, Bohr-Ran; Lee, Li-Ling

2014-01-01

This work reports the color-tunable mixed photoluminescence (PL) emission from an Alq3 organic layer in an Au-Alq3-Au plasmonic structure through the combination of organic fluorescence emission and another form of emission that is enabled by the surface plasmons in the plasmonic structure. The emission wavelength of the latter depends on the Alq3 thickness and can be tuned within the Alq3 fluorescent spectra. Therefore, a two-color broadband, color-tunable mixed PL structure was obtained. Obvious changes in the Commission Internationale d'Eclairage (CIE) coordinates and the corresponding emission colors of Au-Alq3-Au samples clearly varied with the Alq3 thickness (90, 130, and 156 nm).

Color-tunable mixed photoluminescence emission from Alq3 organic layer in metal-Alq3-metal surface plasmon structure

PubMed Central

2014-01-01

This work reports the color-tunable mixed photoluminescence (PL) emission from an Alq3 organic layer in an Au-Alq3-Au plasmonic structure through the combination of organic fluorescence emission and another form of emission that is enabled by the surface plasmons in the plasmonic structure. The emission wavelength of the latter depends on the Alq3 thickness and can be tuned within the Alq3 fluorescent spectra. Therefore, a two-color broadband, color-tunable mixed PL structure was obtained. Obvious changes in the Commission Internationale d’Eclairage (CIE) coordinates and the corresponding emission colors of Au-Alq3-Au samples clearly varied with the Alq3 thickness (90, 130, and 156 nm). PMID:25328506

Depolarization after resonance energy transfer (DARET): a sensitive fluorescence-based assay for botulinum neurotoxin protease activity.

PubMed

Gilmore, Marcella A; Williams, Dudley; Okawa, Yumiko; Holguin, Bret; James, Nicholas G; Ross, Justin A; Roger Aoki, K; Jameson, David M; Steward, Lance E

2011-06-01

The DARET (depolarization after resonance energy transfer) assay is a coupled Förster resonance energy transfer (FRET)-fluorescence polarization assay for botulinum neurotoxin type A or E (BoNT/A or BoNT/E) proteolytic activity that relies on a fully recombinant substrate. The substrate consists of blue fluorescent protein (BFP) and green fluorescent protein (GFP) flanking SNAP-25 (synaptosome-associated protein of 25 kDa) residues 134-206. In this assay, the substrate is excited with polarized light at 387 nm, which primarily excites the BFP, whereas emission from the GFP is monitored at 509 nm. Energy transfer from the BFP to the GFP in the intact substrate results in a substantial depolarization of the GFP emission. The energy transfer is eliminated when the fluorescent domains separate on cleavage by the endopeptidase, and emission from the directly excited GFP product fragment is then highly polarized, resulting in an overall increase in polarization. This increase in polarization can be monitored to assay the proteolytic activity of BoNT/A and BoNT/E in real time. It allows determination of the turnover rate of the substrate and the kinetic constants (V(max) and k(cat)) based on the concentration of cleaved substrate determined directly from the measurements using the additivity properties of polarization. The assay is amenable to high-throughput applications. Copyright © 2011 Elsevier Inc. All rights reserved.

Compound parabolic concentrator optical fiber tip for FRET-based fluorescent sensors

NASA Astrophysics Data System (ADS)

Ul Hassan, Hafeez; Nielsen, Kristian; Aasmul, Soren; Bang, Ole

2015-09-01

The Compound Parabolic Concentrator (CPC) optical fiber tip shape has been proposed for intensity based fluorescent sensors working on the principle of FRET (Förster Resonance Energy Transfer). A simple numerical Zemax model has been used to optimize the CPC tip geometry for a step-index multimode polymer optical fiber for an excitation and emission wavelength of 550 nm and 650nm, respectively. The model suggests an increase of a factor of 1.6 to 4 in the collected fluorescent power for an ideal CPC tip, as compared to the plane-cut fiber tip for fiber lengths between 5 and 45mm.

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

PubMed

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

2014-01-31

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

Tryptophan as key biomarker to detect gastrointestinal tract cancer using non-negative biochemical analysis of native fluorescence and Stokes Shift spectroscopy

NASA Astrophysics Data System (ADS)

Wang, Leana; Zhou, Yan; Liu, Cheng-hui; Zhou, Lixin; He, Yong; Pu, Yang; Nguyen, Thien An; Alfano, Robert R.

2015-03-01

The objective of this study was to find out the emission spectral fingerprints for discrimination of human colorectal and gastric cancer from normal tissue in vitro by applying native fluorescence. The native fluorescence (NFL) and Stokes shift spectra of seventy-two human cancerous and normal colorectal (colon, rectum) and gastric tissues were analyzed using three selected excitation wavelengths (e.g. 300 nm, 320 nm and 340 nm). Three distinct biomarkers, tryptophan, collagen and reduced nicotinamide adenine dinucleotide hydrate (NADH), were found in the samples of cancerous and normal tissues from eighteen subjects. The spectral profiles of tryptophan exhibited a sharp peak in cancerous colon tissues under a 300 nm excitation when compared with normal tissues. The changes in compositions of tryptophan, collagen, and NADH were found between colon cancer and normal tissues under an excitation of 300 nm by the non-negative basic biochemical component analysis (BBCA) model.

Enhanced broadband near-infrared luminescence from Pr3+-doped tellurite glass with silver nanoparticles

NASA Astrophysics Data System (ADS)

Cheng, Pan; Zhou, Yaxun; Zhou, Minghan; Su, Xiue; Zhou, Zizhong; Yang, Gaobo

2017-11-01

Pr3+-doped tellurite glasses containing metallic silver NPs were synthesized by the conventional melt-quenching technique. Structural, thermal and optical properties of the synthesized glass samples were characterized by X-Ray diffraction (XRD) curves, Raman spectra, differential scanning calorimeter (DSC) curves, transmission electron microscopy (TEM) images, UV/Vis/NIR absorption and near-infrared fluorescence emission spectra. The XRD curves confirmed the amorphous structural nature of the synthesized glasses, the Raman spectra identified the presence of different vibrational groups, the DSC curves verified the good thermal stability, and the TEM images revealed the nucleated silver NPs with average diameter about 10 nm dispersed in the glass matrix and its surface Plasmon resonance (SPR) absorption band was located at around 510 nm. Besides, Judd-Ofelt intensity parameters Ωt (t = 2, 4, 6) and other important spectroscopic parameters like transition probability, radiative lifetime, branching ratio were calculated to evaluate the radiative properties of Pr3+ levels from the measured optical absorption spectra. It was found that Pr3+-doped tellurite glasses could emit an ultra-broadband fluorescence extending from 1250 to 1650 nm under the 488 nm excitation, and this fluorescence emission increased further with the introduction of silver NPs. The enhanced fluorescence was mainly attributed to the increased local electric field around Pr3+ induced by silver NPs. The present results demonstrate that Pr3+-Ag codoped tellurite glass is a promising candidate for the near-infrared band ultra-broadband fiber amplifiers covering the expanded low-loss communication window.

Synthesis and properties of Asante Calcium Red--a novel family of long excitation wavelength calcium indicators.

PubMed

Hyrc, Krzysztof L; Minta, Akwasi; Escamilla, P Rogelio; Chan, Patrick P L; Meshik, Xenia A; Goldberg, Mark P

2013-10-01

Although many synthetic calcium indicators are available, a search for compounds with improved characteristics continues. Here, we describe the synthesis and properties of Asante Calcium Red-1 (ACR-1) and its low affinity derivative (ACR-1-LA) created by linking BAPTA to seminaphthofluorescein. The indicators combine a visible light (450-540 nm) excitation with deep-red fluorescence (640 nm). Upon Ca2+ binding, the indicators raise their fluorescence with longer excitation wavelengths producing higher responses. Although the changes occur without any spectral shifts, it is possible to ratio Ca(2+)-dependent (640 nm) and quasi-independent (530 nm) emission when using visible (< 490 nm) or multiphoton (∼780 nm) excitation. Therefore, both probes can be used as single wavelength or, less dynamic, ratiometric indicators. Long indicator emission might allow easy [Ca2+]i measurement in GFP expressing cells. The indicators bind Ca2+ with either high (Kd = 0.49 ± 0.07 μM; ACR-1) or low affinity (Kd = 6.65 ± 0.13 μM; ACR-1-LA). Chelating Zn2+ (Kd = 0.38 ± 0.02 nM) or Mg2+ (Kd∼5mM) slightly raises and binding Co2+ quenches dye fluorescence. New indicators are somewhat pH-sensitive (pKa = 6.31 ± 0.07), but fairly resistant to bleaching. The probes are rather dim, which combined with low AM ester loading efficiency, might complicate in situ imaging. Despite potential drawbacks, ACR-1 and ACR-1-LA are promising new calcium indicators. Copyright © 2013 Elsevier Ltd. All rights reserved.

Synthesis and Properties of Asante Calcium Red –a Novel Family of Long Excitation Wavelength Calcium Indicators

PubMed Central

Hyrc, Krzysztof L.; Minta, Akwasi; Escamilla, P. Rogelio; Chan, Patrick P.L.; Meshik, Xenia A.; Goldberg, Mark P.

2013-01-01

Although many synthetic calcium indicators are available, a search for compounds with improved characteristics continues. Here, we describe the synthesis and properties of Asante Calcium Red-1 (ACR-1) and its low affinity derivative (ACR-1-LA) created by linking BAPTA to seminaphthofluorescein. The indicators combine a visible light (450–540 nm) excitation with deep-red fluorescence (640 nm). Upon Ca2+ binding, the indicators raise their fluorescence with longer excitation wavelengths producing higher responses. Although the changes occur without any spectral shifts, it is possible to ratio Ca2+-dependent (640 nm) and quasi-independent (530 nm) emission when using visible (<490 nm) or multiphoton (~780 nm) excitation. Therefore, both probes can be used as single wavelength or, less dynamic, ratiometric indicators. Long indicator emission might allow easy [Ca2+]i measurement in GFP expressing cells. The indicators bind Ca2+ with either high (Kd=0.49±0.07 μM; ACR-1) or low affinity (Kd=6.65±0.13 μM; ACR-1-LA). Chelating Zn2+ (Kd =0.38±0.02 nM) or Mg2+ (Kd ~5 mM) slightly raises and binding Co2+ quenches dye fluorescence. New indicators are somewhat pH-sensitive (pKa=6.31±0.07), but fairly resistant to bleaching. The probes are rather dim, which combined with low AM ester loading efficiency, might complicate in situ imaging. Despite potential drawbacks, ACR-1 and ACR-1-LA are promising new calcium indicators. PMID:24017967

1.083 μm laser operation in Nd,Mg:LiTaO3 crystal

NASA Astrophysics Data System (ADS)

Hu, P. C.; Hang, Y.; Li, R.; Gong, J.; Yin, J. G.; Zhao, C. C.; He, X. M.; Yu, T.; Zhang, L. H.; Chen, W. B.; Zhu, Y. Y.

2011-10-01

Nd,Mg:LiTaO3 single crystal with high optical quality was grown by Czochralski technique. Absorption and fluorescence spectra were investigated. The peak absorption cross section at 806.5 nm and peak emission cross section at 1091 nm are 6.81×10-20 and 3.28×10-20 cm2, respectively. The fluorescence lifetime was measured to be 129 μs. With a laser-diode as the pump source, a maximum 375 mW continuous-wave laser output at 1083 nm has been obtained with a slope efficiency of 7.2% with respect to the pump power.

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

NASA Astrophysics Data System (ADS)

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

2018-05-01

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

Red electroluminescence and photoluminescence properties of new porphyrin compounds

NASA Astrophysics Data System (ADS)

Zhang, X. H.; Xie, Z. Y.; Wu, F. P.; Zhou, L. L.; Wong, O. Y.; Lee, C. S.; Kwong, H. L.; Lee, S. T.; Wu, S. K.

2003-12-01

Three new porphyrin compounds with saturated red emission were synthesized and their photoluminescence and electroluminescence were studied. The emission of the new dopants peaked at about 635-655 nm and the half-width of the emission peaks was very narrow (below 30 nm). Of the three porphyrin compounds, the one (TBDPP) with the smallest conjugation system had the highest fluorescent quantum yield of 21% in solution. Organic light-emitting diodes using TBDPP as a dopant showed saturated red emission (CIE coordinates of x=0.69, y=0.29) with a luminance of 150 cd/m 2 at a driving voltage of 22 V.

Studies on activated cytostatic fluorouracil as photosensitizer: to use in eye tumor treatment

NASA Astrophysics Data System (ADS)

Pascu, Mihail-Lucian; Carstocea, Benone D.; Brezeanu, Mihail; Voicu, Letitia; Staicu, Angela; Gazdaru, Doina M.; Pascu, Ruxandra A.

2004-09-01

Hydroxypyrimidine 5-fluorouracil (5-FU) belongs to the cytostatics group known as antimetabolites. The effect of UV irradiation on 5-FU was investigated by absorption and fluorescence spectroscopy. The study of the photosensitizer properties of 5-FU was made since their effects could be enhanced by exposure to UV radiation at different doses. Solutions 2.5x10-4M in natural saline water (0.8% NaCl), irradiated with optical beams emitted by N2 laser and UV Hg classic lamp, were used. The 5-FU was chosen due to its strong absorption along a large spectral range which makes possible the fluorescence excitation in UV. The absorption spectra exhibit bands between 250 - 450 nm. The emission fluorescence was measured in the 400-550 nm spectral range, with λex=320 and 350 nm for samples irradiated with Hg lamp and with λex=360 nm for samples irradiated with N2 laser. The excitation fluorescence was measured in the spectral range 200-400 nm, with λem=440 nm for samples irradiated with N2 laser. The spectra reveal a fluorescence enhancement with the exposure time, with a maximum at 3 min due to the transformation of 5-FU molecule into a fluorescent tautomeric form. The destruction more rapid than usual of the neovascularisation was observed for conjunctive of rabbit eyes, when they are impregnated with 5-FU solution and exposed to incoherent UV and visible light.

Determination of the Residual Anthracene Concentration in Cultures of Haloalkalitolerant Actinomycetes by Excitation Fluorescence, Emission Fluorescence, and Synchronous Fluorescence: Comparative Study

PubMed Central

Lara-Severino, Reyna del Carmen; Camacho-López, Miguel Ángel; García-Macedo, Jessica Marlene; Gómez-Oliván, Leobardo M.; Sandoval-Trujillo, Ángel H.; Isaac-Olive, Keila; Ramírez-Durán, Ninfa

2016-01-01

Polycyclic aromatic hydrocarbons (PAHs) are compounds that can be quantified by fluorescence due to their high quantum yield. Haloalkalitolerant bacteria tolerate wide concentration ranges of NaCl and pH. They are potentially useful in the PAHs bioremediation of saline environments. However, it is known that salinity of the sample affects fluorescence signal regardless of the method. The objective of this work was to carry out a comparative study based on the sensitivity, linearity, and detection limits of the excitation, emission, and synchronous fluorescence methods, during the quantification of the residual anthracene concentration from the following haloalkalitolerant actinomycetes cultures Kocuria rosea, Kocuria palustris, Microbacterium testaceum, and 4 strains of Nocardia farcinica, in order to establish the proper fluorescence method to study the PAHs biodegrading capacity of haloalkalitolerant actinobacteria. The study demonstrated statistical differences among the strains and among the fluorescence methods regarding the anthracene residual concentration. The results showed that excitation and emission fluorescence methods performed very similarly but sensitivity in excitation fluorescence is slightly higher. Synchronous fluorescence using Δλ = 150 nm is not the most convenient method. Therefore we propose the excitation fluorescence as the fluorescence method to be used in the study of the PAHs biodegrading capacity of haloalkalitolerant actinomycetes. PMID:26925294

Fluorescence Spectral Properties of All4261 Binding with Phycocyanobilin in E.Coli

NASA Astrophysics Data System (ADS)

Ma, Q.; Zheng, X. J.; Zhou, Z.; Zhou, N.; Zhao, K. H.; Zhou, M.

2014-07-01

Cyanobacteriochromes (CBCRs) are chromophorylated proteins that acting as sensory photoreceptors in cyanobacteria. Based on the bioinformatics of All4261 in Nostoc sp. PCC7120, All4261 is a CBCR apoprotein composed of GAF domains in the N-terminal region. Via polymerase chain reaction with specific primers, All4261 was amplified with genome DNA of Nostoc sp. PCC7120 as template and then subcloned into the expression vector pET30(a+). To survey the fluorescence spectral properties, All4261 was coexpressed with the plasmid that catalyzes phycocyanobilin (PCB) biosynthesis, pACYC-ho1-pcyA, in E.coli BL21. Fluorescence emission spectra and excitation spectra showed that chromophorylated cells containing All4261-PCB had a fluorescence emission peak at 645 nm and a fluorescence excitation peak at 550 nm, but no reversible photoconversion. In order to identify the binding site of PCB in All4261, we obtained three variants All4261(C296L), All4261(C328A), and All4261(C339L), via sitedirected mutagenesis. The binding site was identified as C339 based on the lack of PCB binding of All4261(C339L).

Synthesis of Water Dispersible Fluorescent Carbon Nanocrystals from Syzygium cumini Fruits for the Detection of Fe3+ Ion in Water and Biological Samples and Imaging of Fusarium avenaceum Cells.

PubMed

Bhamore, Jigna R; Jha, Sanjay; Singhal, Rakesh Kumar; Kailasa, Suresh Kumar

2017-01-01

In this work, water dispersible fluorescent carbon nanocrystals (NCs) were synthesized by a simple, green and low cost hydrothermal method using Syzygium cumini (jamun) as a carbon source at 180 °C for 6 h. The average size of carbon NCs was found to be 2.1 ± 0.5 nm and shown bright blue fluorescence when excited at 365 nm under UV lamp. The carbon NCs were characterized by spectroscopic (UV-visible and fluorescence, Fourier transform infrared and dynamic light scattering) and high resolution transmission electron microscopic techniques. The quantum yield of carbon NCs was found to be ~5.9 % at 438 nm emission wavelength when excited at 360 nm. It was noticed that none of the metal ions quenched the fluorescence intensity of carbon NCs at 438 nm except for Fe 3+ , indicating the formation of Fe 3+ ion-carbon NCs complexes. The linear range was observed in the concentration range of 0.01-100 μM with the corresponding detection limits of 0.001 μM, respectively. Furthermore, the carbon NCs were used as probes for imaging of fungal (Fusarium avenaceum) cells.

Detecting beta-amyloid aggregation from time-resolved emission spectra

NASA Astrophysics Data System (ADS)

Alghamdi, A.; Vyshemirsky, V.; Birch, D. J. S.; Rolinski, O. J.

2018-04-01

The aggregation of beta-amyloids is one of the key processes responsible for the development of Alzheimer’s disease. Early molecular-level detection of beta-amyloid oligomers may help in early diagnosis and in the development of new intervention therapies. Our previous studies on the changes in beta-amyloid’s single tyrosine intrinsic fluorescence response during aggregation demonstrated a four-exponential fluorescence intensity decay, and the ratio of the pre-exponential factors indicated the extent of the aggregation in the early stages of the process before the beta-sheets were formed. Here we present a complementary approach based on the time-resolved emission spectra (TRES) of amyloid’s tyrosine excited at 279 nm and fluorescence in the window 240-450 nm. TRES have been used to demonstrate sturctural changes occuring on the nanosecond time scale after excitation which has significant advantages over using steady-state spectra. We demonstrate this by resolving the fluorescent species and revealing that beta-amyloid’s monomers show very fast dielectric relaxation, and its oligomers display a substantial spectral shift due to dielectric relaxation, which gradually decreases when the oligomers become larger.

Probing the conformational dynamics of photosystem I in unconfined and confined spaces.

PubMed

Das, Gaurav; Chattoraj, Shyamtanu; Nandi, Somen; Mondal, Prasenjit; Saha, Abhijit; Bhattacharyya, Kankan; Ghosh, Surajit

2017-12-20

The fluorescence dynamics of Photosystem I (PSI) in bulk water and inside a confined environment like a liposome have been investigated using time resolved confocal microscopy. In bulk water, PSI exhibits a major emission peak at ∼680 nm, while in the liposome it exhibits a markedly blue shifted emission maximum at ∼485 nm. This is indicative of conformational changes due to entrapment and emergence of a stressed conformation of PSI inside the liposome. The observed time constants for the fluorescence lifetime of PSI inside the liposome are significantly high as opposed to PSI in bulk water. More interestingly, the fluorescence intensity of PSI in bulk water exhibits strong fluctuations with many high intensity jumps and these are anti-correlated with the fluorescence lifetime of PSI. In contrast, inside the liposome, no such anti-correlated behaviour is observed. We further demonstrated that PSI exhibits at least two conformational states in bulk water, whereas a single conformation is observed inside the liposome, indicating the conformational rigidity and locking of the PSI complex inside a liposome.

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

NASA Astrophysics Data System (ADS)

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

2006-05-01

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

Visualization of melanoma tumor with lectin-conjugated rare-earth doped fluoride nanocrystals

PubMed Central

Dumych, Tetiana; Lutsyk, Maxym; Banski, Mateusz; Yashchenko, Antonina; Sojka, Bartlomiej; Horbay, Rostyslav; Lutsyk, Alexander; Stoika, Rostyslav; Misiewicz, Jan; Podhorodecki, Artur; Bilyy, Rostyslav

2014-01-01

Aim To develop specific fluorescent markers for melanoma tumor visualization, which would provide high selectivity and reversible binding pattern, by the use of carbohydrate-recognizing proteins, lectins, combined with the physical ability for imaging deep in the living tissues by utilizing red and near infrared fluorescent properties of specific rare-earth doped nanocrystals (NC). Methods B10F16 melanoma cells were inoculated to C57BL/6 mice for inducing experimental melanoma tumor. Tumors were removed and analyzed by lectin-histochemistry using LABA, PFA, PNA, HPA, SNA, GNA, and NPL lectins and stained with hematoxylin and eosin. NPL lectin was conjugated to fluorescent NaGdF4:Eu3+-COOH nanoparticles (5 nm) via zero length cross-linking reaction, and the conjugates were purified from unbound substances and then used for further visualization of histological samples. Fluorescent microscopy was used to visualize NPL-NaGdF4:Eu3+ with the fluorescent emission at 600-720 nm range. Results NPL lectin selectively recognized regions of undifferentiated melanoblasts surrounding neoangiogenic foci inside melanoma tumor, PNA lectin recognized differentiated melanoblasts, and LCA and WGA were bound to tumor stroma regions. NPL-NaGdF4:Eu3+ conjugated NC were efficiently detecting newly formed regions of melanoma tumor, confirmed by fluorescent microscopy in visible and near infrared mode. These conjugates possessed high photostability and were compatible with convenient xylene-based mounting systems and preserved intensive fluorescent signal at samples storage for at least 6 months. Conclusion NPL lectin-NaGdF4:Eu3+ conjugated NC permitted distinct identification of contours of the melanoma tissue on histological sections using red excitation at 590-610 nm and near infrared emission of 700-720 nm. These data are of potential practical significance for development of glycans-conjugated nanoparticles to be used for in vivo visualization of melanoma tumor. PMID:24891277

Selective detection of Mg2+ ions via enhanced fluorescence emission using Au–DNA nanocomposites

PubMed Central

Basu, Tanushree; Rana, Khyati; Das, Niranjan

2017-01-01

The biophysical properties of DNA-modified Au nanoparticles (AuNPs) have attracted a great deal of research interest for various applications in biosensing. AuNPs have strong binding capability to the phosphate and sugar groups in DNA, rendering unique physicochemical properties for detection of metal ions. The formation of Au–DNA nanocomposites is evident from the observed changes in the optical absorption, plasmon band, zeta potential, DLS particle size distribution, as well as TEM and AFM surface morphology analysis. Circular dichroism studies also revealed that DNA-functionalized AuNP binding caused a conformational change in the DNA structure. Due to the size and shape dependent plasmonic interactions of AuNPs (33–78 nm) with DNA, the resultant Au–DNA nanocomposites (NCs) exhibit superior fluorescence emission due to chemical binding with Ca2+, Fe2+ and Mg2+ ions. A significant increase in fluorescence emission (λex = 260 nm) of Au–DNA NCs was observed after selectively binding with Mg2+ ions (20–800 ppm) in an aqueous solution where a minimum of 100 ppm Mg2+ ions was detected based on the linearity of concentration versus fluorescence intensity curve (λem = 400 nm). The effectiveness of Au–DNA nanocomposites was further verified by comparing the known concentration (50–120 ppm) of Mg2+ ions in synthetic tap water and a real life sample of Gelusil (300–360 ppm Mg2+), a widely used antacid medicine. Therefore, this method could be a sensitive tool for the estimation of water hardness after careful preparation of a suitably designed Au–DNA nanostructure. PMID:28487819

Synthesis of silver nanoparticles stabilized with C-phycocyanin and for fluorimetric detection of copper ions

NASA Astrophysics Data System (ADS)

Wei, Nana; Hou, Yanhua; Lu, Zongbao; Yu, Huatong; Wang, Quanfu

2018-01-01

In this study, C-phycocyanin as protective agent, AgNO3 as raw material and NaBH4 as reducing agent synthesized C-phycocyanin-Ag nanoparticles (PC-AgNPs). The synthesis conditions of PC-AgNPs were determined by optimization. The maximum UV absorption peak of PC-AgNPs at 400 nm. The fluorescence excitation wavelength was 580 nm and the emission wavelength was 625 nm. PC-AgNPs was spherical in transmission electron microscope and the particles sizes were about 10-25 nm. In addition, fluorescence quenching was observed after adding copper ions to PC-AgNPs, which indicated that PC-AgNPs has potential applications in the detection of copper ions in diverse water environment.

A novel fluorescent aptasensor based on gold and silica nanoparticles for the ultrasensitive detection of ochratoxin A

NASA Astrophysics Data System (ADS)

Taghdisi, Seyed Mohammad; Danesh, Noor Mohammad; Beheshti, Hamed Reza; Ramezani, Mohammad; Abnous, Khalil

2016-02-01

Analytical approaches for the detection and quantitation of ochratoxin A (OTA) in blood serum and food products are high in demand. In this study, a fluorescent aptamer-based sensor (aptasensor) is developed for the selective and sensitive detection of OTA, based on a complementary strand of aptamer (CS) and two types of nanoparticles, gold nanoparticles (AuNPs) and silica nanoparticles (SNPs) coated with streptavidin. The fabricated aptasensor inherits the characteristics of SNPs, as enhancers of fluorescence intensity; AuNPs, such as large surface area and unique optical properties; and high affinity of the aptamer toward its target compared to its CS. In the absence of OTA, no FAM and biotin-labeled CS is in the environment of the SNPs coated with streptavidin, which leads to no fluorescence emission. In the presence of the target, an FAM and biotin-labeled CS-SNPs coated with streptavidin conjugate is formed, thus resulting in a very strong fluorescence emission. The designed fluorescent aptasensor exhibits high selectivity toward OTA with a limit of detection (LOD) as low as 0.098 nM. Furthermore, the fabricated aptasensor was successfully applied for the detection of OTA in grape juice and serum with LODs of 0.113 and 0.152 nM, respectively.

Polythiophene nanofilms for sensitive fluorescence detection of viruses in drinking water.

PubMed

Wankar, Shashwati; Turner, Nicholas W; Krupadam, Reddithota J

2016-08-15

Molecular imprints of the tobacco necrosis virus (TNV) have been formed within polythiophene nanofilms with an approximate thickness of 200nm. These films have been electrochemically deposited onto conducting Au surfaces. Upon rebinding, the TNV-polythiophene complex changes the fluorescence intensity of the nanofilm. The fluorescence intensity at 410nm was observed to be proportional to the concentration of viruses in the range of 0.1-10ngL(-1) (0.15-15pg) with the lower calculated detection limit of 2.29ngL(-1) (3.4pg). The intensity of the fluorescence emission is not affected by the thickness of the polythiophene film and the nature of TNV specific binding sites. Kinetic data analyses showed that the nanofilm responds to TNV within 2min; and cross-selectivity studies with tobacco mosaic virus (TMV) showed an excellent specificity for the targeted TNV. These binding experiments demonstrate the potential of fluorescence emission for the specific, label free and rapid detection of viruses using nanofilm sensors. Taking into account the lower limit of detection, the fluorescence sensing reported here is reliable, simple to perform, rapid, cost-effective and offers a sensitive analytical method for virus detection in water resources. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluorescence spectroscopy of Rhodamine 6G: concentration and solvent effects.

PubMed

Zehentbauer, Florian M; Moretto, Claudia; Stephen, Ryan; Thevar, Thangavel; Gilchrist, John R; Pokrajac, Dubravka; Richard, Katherine L; Kiefer, Johannes

2014-01-01

Rhodamine 6G (R6G), also known as Rhodamine 590, is one of the most frequently used dyes for application in dye lasers and as a fluorescence tracer, e.g., in the area of environmental hydraulics. Knowing the spectroscopic characteristics of the optical emission is key to obtaining high conversion efficiency and measurement accuracy, respectively. In this work, solvent and concentration effects are studied. A series of eight different organic solvents (methanol, ethanol, n-propanol, iso-propanol, n-butanol, n-pentanol, acetone, and dimethyl sulfoxide (DMSO)) are investigated at constant dye concentration. Relatively small changes of the fluorescence spectrum are observed for the different solvents; the highest fluorescence intensity is observed for methanol and lowest for DMSO. The shortest peak wavelength is found in methanol (568 nm) and the longest in DMSO (579 nm). Concentration effects in aqueous R6G solutions are studied over the full concentration range from the solubility limit to highly dilute states. Changing the dye concentration provides tunability between ∼550 nm in the dilute case and ∼620 nm at high concentration, at which point the fluorescence spectrum indicates the formation of R6G aggregates. Copyright © 2013 Elsevier B.V. All rights reserved.

Dye-enhanced reflectance and fluorescence confocal microscopy as an optical pathology tool

NASA Astrophysics Data System (ADS)

Yaroslavsky, Anna N.; Salomatina, Elena; Novak, John; Amat-Roldan, Ivan; Castano, Ana; Hamblin, Michael

2006-02-01

Early detection and precise excision of neoplasms are imperative requirements for successful cancer treatment. In this study we evaluated the use of dye-enhanced confocal microscopy as an optical pathology tool in the ex vivo trial with fresh thick non-melanoma skin cancer excisions and in vivo trial with B16F10 melanoma cancer in mice. For the experiments the tumors were rapidly stained using aqueous solutions of either toluidine blue or methylene blue and imaged using multimodal confocal microscope. Reflectance images were acquired at the wavelengths of 630nm and 650 nm. Fluorescence was excited at 630 nm and 650 nm. Fluorescence emission was registered in the range between 680 nm and 710 nm. The images were compared to the corresponding en face frozen H&E sections. The results of the study indicate confocal images of stained cancerous tissue closely resemble corresponding H&E sections both in vivo and in vitro. This remarkable similarity enables interpretation of confocal images in a manner similar to that of histopathology. The developed technique may provide an efficient real-time optical tool for detecting skin pathology.

Monitoring biological aerosols using UV fluorescence

NASA Astrophysics Data System (ADS)

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

1999-01-01

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

Determination of ethambutol by a sensitive fluorescent probe

NASA Astrophysics Data System (ADS)

Wu, Wen-Ying; Yang, Ji-Yuan; Du, Li-Ming; Wu, Hao; Li, Chang-Feng

2011-08-01

The competitive reaction between ethambutol and two fluorescent probes (i.e., berberine and palmatine) for occupancy of the cucurbit[7]uril (CB[7]) cavity was studied by spectrofluorometry. The CB[7] reacts with these probes to form stable complexes, and the fluorescence intensity of the complexes is greatly enhanced. In addition, the excitation and emission wavelengths of their complexes moved to wavelengths of 343 nm and 495 nm, respectively. However, the addition of ethambutol dramatically quenches the fluorescence intensity of the two complexes. Accordingly, a couple of new fluorescence quenching methods for the determination of ethambutol were established. The methods can be applied for quantifying ethambutol. A linear relationship between the fluorescence quenching values (Δ F) and ethambutol concentration exists in the range of 5.0-1000.0 ng mL -1, with a correlation coefficient ( r) of 0.9997. The detection limit is 1.7 ng mL -1. The fluorescent probe of berberine has higher sensitivity than palmatine. This paper also discusses the mechanism of fluorescence indicator probes.

Glucose sensing molecules having selected fluorescent properties

DOEpatents

Satcher, Jr., Joe H.; Lane, Stephen M.; Darrow, Christopher B.; Cary, Douglas R.; Tran, Joe Anh

2004-01-27

An analyte sensing fluorescent molecule that employs intramolecular electron transfer is designed to exhibit selected fluorescent properties in the presence of analytes such as saccharides. The selected fluorescent properties include excitation wavelength, emission wavelength, fluorescence lifetime, quantum yield, photostability, solubility, and temperature or pH sensitivity. The compound comprises an aryl or a substituted phenyl boronic acid that acts as a substrate recognition component, a fluorescence switch component, and a fluorophore. The fluorophore and switch component are selected such that the value of the free energy for electron transfer is less than about 3.0 kcal mol.sup.-1. Fluorescent compounds are described that are excited at wavelengths greater than 400 nm and emit at wavelengths greater than 450 nm, which is advantageous for optical transmission through skin. The fluorophore is typically selected from transition metal-ligand complexes and thiazine, oxazine, oxazone, or oxazine-one as well as anthracene compounds. The fluorescent compound can be immobilized in a glucose permeable biocompatible polymer matrix that is implantable below the skin.

Detection of silver nanoparticles in cells by flow cytometry using light scatter and far-red fluorescence.

PubMed

Zucker, R M; Daniel, K M; Massaro, E J; Karafas, S J; Degn, L L; Boyes, W K

2013-10-01

The cellular uptake of different sized silver nanoparticles (AgNP) (10, 50, and 75 nm) coated with polyvinylpyrrolidone (PVP) or citrate on a human derived retinal pigment epithelial cell line (ARPE-19) was detected by flow cytometry following 24-h incubation of the cells with AgNP. A dose dependent increase of side scatter and far red fluorescence was observed with both PVP and citrate-coated 50 nm or 75 nm silver particles. Using five different flow cytometers, a far red fluorescence signal in the 700-800 nm range increased as much as 100 times background as a ratio comparing the intensity measurements of treated sample and controls. The citrate-coated silver nanoparticles (AgNP) revealed slightly more side scatter and far red fluorescence than did the PVP coated silver nanoparticles. This increased far red fluorescence signal was observed with 50 and 75 nm particles, but not with 10 nm particles. Morphological evaluation by dark field microscopy showed silver particles (50 and 75 nm) clumped and concentrated around the nucleus. One possible hypothesis to explain the emission of far red fluorescence from cells incubated with silver nanoparticles is that the silver nanoparticles inside cells agglomerate into small nano clusters that form surface plasmon resonance which interacts with laser light to emit a strong far red fluorescence signal. The results demonstrate that two different parameters (side scatter and far red fluorescence) on standard flow cytometers can be used to detect and observe metallic nanoparticles inside cells. The strength of the far red fluorescence suggests that it may be particularly useful for applications that require high sensitivity. © Published 2013 Wiley-Periodicals, Inc. Published 2013 Wiley‐Periodicals, Inc. This article is a US government work and, as such, is in the public domain in the United States of America.

Hydroxylated near-infrared BODIPY fluorophores as intracellular pH sensors

PubMed Central

Salim, Mohamed M.; Owens, Eric A.; Gao, Tielong; Lee, Jeong Heon; Hyun, Hoon; Choi, Hak Soo; Henary, Maged

2015-01-01

In this study, a series of new, highly sensitive BF2-chelated tetraarylazadipyrromethane dyes are synthesized and analyzed to be suitable as on/off photo-induced electron transfer modulated fluorescent sensors for determination of intracellular pH. The ethanolic solutions of the new indicators feature absorption maxima in the range of 696–700 nm and a fluorescence emission maximum at 720 nm. Molar absorptivity and fluorescence quantum yield data were determined for the studied set of aza-BODIPY indicators. These indicators have high molar absorption coefficients of ~80 000 M−1 cm−1 and quantum yields (up to 18%). Corresponding pKa values of indicators are determined from absorbance and fluorescence measurements and range from 9.1 to 10.8, depending on the selective positioning of electron-donating functionalities. The excellent photostability of the aza-BODIPY indicators makes them particularly suitable for long duration measurements. The in vitro cellular staining of living tissues in PC3 cells based on the isosbestic point at pH 7.8 and pH 9.3 has been employed which shows an increase in fluorescence intensity at 800 nm with increase in pH for certain compounds and fluorescence intensity decreases at 700 nm. Therefore, the new indicators are suitable for exploitation and adaptation in a diverse range of analytical applications. PMID:25105177

Synchronous fluorescence spectroscopy of colon neoplasia

NASA Astrophysics Data System (ADS)

Borisova, Ekaterina; Semyachkina-Glushkovskaya, Oxana; Genova, Tsanislava; Penkov, Nikolay; Terziev, Ivan; Vladimirov, Borislav; Avramov, Latchezar

2017-03-01

Synchronous fluorescence spectroscopy (SFS) is a steady-state approach that we used for evaluation of specific fluorescence characteristics of cancerous colorectal tissues. SFS allow narrowing of the fluorescence spectra received, which increase the spectral resolution and improve the analysis of the fluorescence origin in such complex objects, such as biological tissues. In our study we investigate the characteristic differences, with diagnostic meaning, in the synchronous fluorescence spectra (SFS) of cancerous and healthy colorectal tissues ex vivo using a spectrofluorimeter FluoroLog3 (HORIBA, JobinYvon, France) for obtaining of the SFS data in a broad spectral range (300-800 nm) using excitation in the range of 280-440 nm with a delta lambda between 0 and 200 nm with a 10 nm step between scanning excitation and emission data. The procedure of obtaining the investigated samples ex vivo includes their excision during surgery for removal of neoplasia lesions. After the surgical removal biological samples are transported in isothermal conditions and safekeeping solution from the hospital to the spectral laboratory, where their spectral properties were investigated. All patients received and signed written informed consent and this research is approved by Ethics committee of University Hospital "Tsaritsa Yoanna", Sofia. Histological analysis was used as "gold standard" for evaluation of tissue samples and comparison of the spectral data received.

Application of fluorescent Eu:Gd2O3 nanoparticles to the visualization of protein micropatterns

NASA Astrophysics Data System (ADS)

Dosev, Dosi; Nichkova, Mikaela; Liu, Maozi; Guo, Bing; Liu, Gang-yu; Xia, Younan; Hammock, Bruce D.; Kennedy, Ian M.

2005-03-01

Nanoparticles made of lanthanide oxides are promising fluorophores as a new class of tags in biochemistry because of their large Stokes shift, sharp emission spectra, long lifetime and lack of photobleaching. We demonstrate for first time the application of these nanoparticles to the visualization of protein micropatterns. Europium-doped gadolinium oxide (Eu:Gd2O3) nanoparticles were synthesized by spray pyrolysis and were characterized by means of laser-induced fluorescent spectroscopy and TEM. Their main emission peak is at 612 nm. And their size distribution is from 5 nm to 500 nm. The nanoparticles were coated with avidin through physical adsorption. Biotinylated Bovine Serum Albumin (BSA-b) was patterned on a silicon wafer using a micro-contact printing technique. The BSA-b - patterned wafer was incubated in a solution containing the avidin-coated nanoparticles. The specific interaction between biotin and avidin was studied by means of fluorescent microscopy and atomic-force microscopy (AFM). The fluorescent microscopic images revealed that the nanoparticles were organized into designated structures as defined by the microcontact printing process - non-specific binding of the avidin-coated nanoparticles to bare substrate was negligible. The fluorescent pattern did not suffer any photobleaching during the observation process which demonstrates the suitability of Eu:Gd2O3 nanoparticles as fluorescent labels with extended excitation periods - organic dyes, including chelates, suffer bleaching over the same period. More detailed studies were preformed using AFM at a single nanoparticle level. The specific and the non-specific binding densities of the particles were qualitatively evaluated.

Smart textile framework: Photochromic and fluorescent cellulosic fabric printed by strontium aluminate pigment.

PubMed

Khattab, Tawfik A; Rehan, Mohamed; Hamouda, Tamer

2018-09-01

Smart clothing can be defined as textiles that respond to a certain stimulus accompanied by a change in their properties. A specific class herein is the photochromic and fluorescent textiles that change color with light. A photochromic and fluorescent cotton fabric based on pigment printing is obtained. Such fabric is prepared by aqueous-based pigment-binder printing formulation containing inorganic pigment phosphor characterized by good photo- and thermal stability. It exhibits optimal excitation wavelength (365 nm) results in color and fluorescence change of the fabric surface. To prepare the transparent pigment-binder composite film, the phosphor pigment must be well-dispersed via physical immobilization without their aggregation. The pigment-binder paste is applied successfully onto cotton fabric using screen printing technique followed by thermal fixation. After screen-printing, a homogenous photochromic film is assembled on a cotton substrate surface, which represents substantial greenish-yellow color development as indicated by CIE Lab color space measurements under ultraviolet light, even at a pigment concentration of 0.08 wt% of the printing paste. The photochromic cotton fabric exhibit three excitation peaks at 272, 325 and 365 nm and three emission peaks at 418, 495 and 520 nm. The fluorescent optical microscope, scanning electron microscope, elemental mapping, energy dispersive X-ray spectroscopy, fluorescence emission and UV/Vis absorption spectroscopic data of the printed cotton fabric are described. The printed fabric showed a reversible and rapid photochromic response during ultra-violet excitation without fatigue. The fastness properties including washing, crocking, perspiration, sublimation/heat, and light are described. Copyright © 2018 Elsevier Ltd. All rights reserved.

Extended Stokes shift in fluorescent proteins: chromophore-protein interactions in a near-infrared TagRFP675 variant.

PubMed

Piatkevich, Kiryl D; Malashkevich, Vladimir N; Morozova, Kateryna S; Nemkovich, Nicolai A; Almo, Steven C; Verkhusha, Vladislav V

2013-01-01

Most GFP-like fluorescent proteins exhibit small Stokes shifts (10-45 nm) due to rigidity of the chromophore environment that excludes non-fluorescent relaxation to a ground state. An unusual near-infrared derivative of the red fluorescent protein mKate, named TagRFP675, exhibits the Stokes shift, which is 30 nm extended comparing to that of the parental protein. In physiological conditions, TagRFP675 absorbs at 598 nm and emits at 675 nm that makes it the most red-shifted protein of the GFP-like protein family. In addition, its emission maximum strongly depends on the excitation wavelength. Structures of TagRFP675 revealed the common DsRed-like chromophore, which, however, interacts with the protein matrix via an extensive network of hydrogen bonds capable of large flexibility. Based on the spectroscopic, biochemical, and structural analysis we suggest that the rearrangement of the hydrogen bond interactions between the chromophore and the protein matrix is responsible for the TagRFP675 spectral properties.

Hyperspectral Image Analysis for Skin Tumor Detection

NASA Astrophysics Data System (ADS)

Kong, Seong G.; Park, Lae-Jeong

This chapter presents hyperspectral imaging of fluorescence for nonin-vasive detection of tumorous tissue on mouse skin. Hyperspectral imaging sensors collect two-dimensional (2D) image data of an object in a number of narrow, adjacent spectral bands. This high-resolution measurement of spectral information reveals a continuous emission spectrum for each image pixel useful for skin tumor detection. The hyperspectral image data used in this study are fluorescence intensities of a mouse sample consisting of 21 spectral bands in the visible spectrum of wavelengths ranging from 440 to 640 nm. Fluorescence signals are measured using a laser excitation source with the center wavelength of 337 nm. An acousto-optic tunable filter is used to capture individual spectral band images at a 10-nm resolution. All spectral band images are spatially registered with the reference band image at 490 nm to obtain exact pixel correspondences by compensating the offsets caused during the image capture procedure. The support vector machines with polynomial kernel functions provide decision boundaries with a maximum separation margin to classify malignant tumor and normal tissue from the observed fluorescence spectral signatures for skin tumor detection.

Application of normal fluorescence and stability-indicating derivative synchronous fluorescence spectroscopy for the determination of gliquidone in presence of its fluorescent alkaline degradation product

NASA Astrophysics Data System (ADS)

El-ghobashy, Mohamed R.; Yehia, Ali M.; Helmy, Aya H.; Youssef, Nadia F.

2018-01-01

Simple, smart and sensitive normal fluorescence and stability-indicating derivative synchronous spectrofluorimetric methods have been developed and validated for the determination of gliquidone in the drug substance and drug product. Normal spectrofluorimetric method of gliquidone was established in methanol at λ excitation 225 nm and λ emission 400 nm in concentration range 0.2-3 μg/ml with LOD equal 0.028. The fluorescence quantum yield of gliquidone was calculated using quinine sulfate as a reference and found to be 0.542. Stability-indicating first and third derivative synchronous fluorescence spectroscopy were successfully utilized to overcome the overlapped spectra in normal fluorescence of gliquidone and its alkaline degradation product. Derivative synchronous methods are based on using the synchronous fluorescence of gliquidone and its degradation product in methanol at Δ λ50 nm. Peak amplitude in the first derivative of synchronous fluorescence spectra was measured at 309 nm where degradation product showed zero-crossing without interference. The peak amplitudes in the third derivative of synchronous fluorescence spectra, peak to trough were measured at 316,329 nm where degradation product showed zero-crossing. The different experimental parameters affecting the normal and synchronous fluorescence intensity of gliquidone were studied and optimized. Moreover, the cited methods have been validated as per ICH guidelines. The peak amplitude-concentration plots of the derivative synchronous fluorescence were linear over the concentration range 0.05-2 μg/ml for gliquidone. Limits of detection were 0.020 and 0.022 in first and third derivative synchronous spectra, respectively. The adopted methods were successfully applied to commercial tablets and the results demonstrated that the derivative synchronous fluorescence spectroscopy is a powerful stability-indicating method, suitable for routine use with a short analysis time. Statistical comparison between the results obtained by normal fluorescence and derivative synchronous methods and the official one using student's t-test and F-ratio showed no significant difference regarding accuracy and precision.

Time-domain laser-induced fluorescence spectroscopy apparatus for clinical diagnostics

NASA Astrophysics Data System (ADS)

Fang, Qiyin; Papaioannou, Thanassis; Jo, Javier A.; Vaitha, Russel; Shastry, Kumar; Marcu, Laura

2004-01-01

We report the design and development of a compact optical fiber-based apparatus for in situ time-resolved laser-induced fluorescence spectroscopy (tr-LIFS) of biological systems. The apparatus is modular, optically robust, and compatible with the clinical environment. It incorporates a dual output imaging spectrograph, a gated multichannel plate photomultiplier (MCP-PMT), an intensified charge-coupled-device (ICCD) camera, and a fast digitizer. It can accommodate various types of light sources and optical fiber probes for selective excitation and remote light delivery/collection as required by different applications. The apparatus allows direct recording of the entire fluorescence decay with high sensitivity (nM range fluorescein dye concentration with signal-to-noise ratio of 46) and with four decades dynamic range. It is capable of resolving a broad range of fluorescence lifetimes from hundreds of picoseconds (as low as 300 ps) using the MCP-PMT coupled to the digitizer to milliseconds using the ICCD. The data acquisition and analysis process is fully automated, enabling fast recording of fluorescence intensity decay across the entire emission spectrum (0.8 s per wavelength or ˜40 s for a 200 nm wavelength range at 5 nm increments). The spectral and temporal responses of the apparatus were calibrated and its performance was validated using fluorescence lifetime standard dyes (Rhodamin B, 9-cyanoanthracene, and rose Bengal) and tissue endogenous fluorophores (elastin, collagen, nicotinamide adenine dinucleotide, and flavin adenine dinucleotide). Fluorescence decay lifetimes and emission spectra of all tested compounds measured with the current tr-LIFS apparatus were found in good agreement with the values reported in the literature. The design and performance of tr-LIFS apparatus have enabled in vivo studies of atherosclerotic plaques and brain tumors.

Autofluorescence polarization spectroscopy of cancerous and normal colorectal tissues

NASA Astrophysics Data System (ADS)

Genova, Ts.; Borisova, E.; Penkov, N.; Vladimirov, B.; Terziev, I.; Zhelyazkova, Al.; Avramov, L.

2016-01-01

The wide spread of colorectal cancer and high mortality rate among the patients, brings it to a level of high public health concern. Implementation of standard endoscopic surveillance proves to be effective for reduction of colorectal cancer patients' mortality, since its early diagnosis allows eradication of the disease prior to invasive cancer development, but its application in common clinical practice is still limited. Therefore the development of complimentary diagnostic techniques of the standard white-light endoscopy is on high demand. The non-invasive and highly informative nature of the fluorescence spectroscopy allow to use it as the most realistic prospect of an add-on "red flag" technique for early endoscopy detection of colorectal cancer. Synchronous fluorescence spectroscopy (SFS) is a steady-state approach that is used for evaluation of specific fluorescence characteristics of cancerous colorectal tissues in our studies. The feasibility of polarization fluorescence technique to enhance the contrast between normal and cancerous tissues was investigated as well. Additional linear polarizing optics was used on the way of the excitation and emission fluorescence light beams. The polarizing effects were investigated in parallel and perpendicular linear polarization modes respectively. The excitation applied was in the region of 280 - 440 nm, with 10 nm scanning step, and the fluorescence emission was detected in the region of 300 - 800 nm. Our previous experience with SFS technique showed its great potential for accurate, highly sensitive and specific discrimination between cancerous and normal colorectal tissue. Since one of the major sources of endogenous fluorescence with diagnostic meaning is the structural protein - collagen, which is characterized with high anisotropy, we've expected and observed an enhancement of the spectral differences between cancerous and normal colorectal tissue, which could be beneficial for the colorectal tumour' diagnostics using SFS.

Remote sensing of nutrient deficiency in Lactuca sativa using neural networks for terrestrial and advanced life support applications

NASA Astrophysics Data System (ADS)

Sears, Edie Seldon

2000-12-01

A remote sensing study using reflectance and fluorescence spectra of hydroponically grown Lactuca sativa (lettuce) canopies was conducted. An optical receiver was designed and constructed to interface with a commercial fiber optic spectrometer for data acquisition. Optical parameters were varied to determine effects of field of view and distance to target on vegetation stress assessment over the test plant growth cycle. Feedforward backpropagation neural networks (NN) were implemented to predict the presence of canopy stress. Effects of spatial and spectral resolutions on stress predictions of the neural network were also examined. Visual inspection and fresh mass values failed to differentiate among controls, plants cultivated with 25% of the recommended concentration of phosphorous (P), and those cultivated with 25% nitrogen (N) based on fresh mass and visual inspection. The NN's were trained on input vectors created using reflectance and test day, fluorescence and test day, and reflectance, fluorescence, and test day. Four networks were created representing four levels of spectral resolution: 100-nm NN, 10-nm NN, 1-nm NN, and 0.1-nm NN. The 10-nm resolution was found to be sufficient for classifying extreme nitrogen deficiency in freestanding hydroponic lettuce. As a result of leaf angle and canopy structure broadband scattering intensity in the 700-nm to 1000-nm range was found to be the most useful portion of the spectrum in this study. More subtle effects of "greenness" and fluorescence emission were believed to be obscured by canopy structure and leaf orientation. As field of view was not as found to be as significant as originally believed, systems implementing higher repetitions over more uniformly oriented, i.e. smaller, flatter, target areas would provide for more discernible neural network input vectors. It is believed that this technique holds considerable promise for early detection of extreme nitrogen deficiency. Further research is recommended using stereoscopic digital cameras to quantify leaf area index, leaf shape, and leaf orientation as well as reflectance. Given this additional information fluorescence emission may also prove a more useful biological assay of freestanding vegetation.

Native fluorescence spectroscopy of blood plasma of rats with experimental diabetes: identifying fingerprints of glucose-related metabolic pathways

NASA Astrophysics Data System (ADS)

Shirshin, Evgeny; Cherkasova, Olga; Tikhonova, Tatiana; Berlovskaya, Elena; Priezzhev, Alexander; Fadeev, Victor

2015-05-01

We present the results of a native fluorescence spectroscopy study of blood plasma of rats with experimental diabetes. It was shown that the fluorescence emission band shape at 320 nm excitation is the most indicative of hyperglycemia in the blood plasma samples. We provide the interpretation of this fact based on the changes in reduced nicotinamide adenine dinucleotide phosphate concentration due to glucose-related metabolic pathways and protein fluorescent cross-linking formation following nonenzymatic glycation.

Characterization of marine macroalgae by fluorescence signatures

NASA Technical Reports Server (NTRS)

Topinka, J. A.; Bellows, W. Korjeff; Yentsch, C. S.

1990-01-01

The feasibility of distinguishing macroalgal classes by their fluorescence signatures was investigated using narrow-waveband light to excite groups of accessory pigments in brown, red, and green macroalgae and measuring fluorescence emission at 685 nm. Results obtained on 20 marine macroalgae field-collected samples showed that fluorescence excitation signatures were relatively uniform within phylogenetic classes but were substantially different for different classes. It is suggested that it may be possible to characterize the type and the abundance of subtidal macroalgae from low-flying aircraft using existing laser-induced fluorescence methodology.

Emission Behavior of Fluorescently Labeled Silver Nanoshell: Enhanced Self-Quenching by Metal Nanostructure.

PubMed

Zhang, Jian; Fu, Yi; Lakowicz, Joseph R

2007-02-08

Labeled silica beads with an average diameter of 100 nm were synthesized by incorporating with 20-600 μM Ru(bpy)(3) (2+) complexes. Silver shells were deposited on the beads layer-by-layer with the shell thickness of 5-50 nm. The emission band became narrower and the intensity was enhanced depending on the shell thickness. Self-quenching of the probe was observed at high concentration. Poisson statistics were employed to analyze self-quenching of the fluorophores. The estimated quenching distance was extended from 6 to 16 nm with shell growth from 0 to 50 nm. Moreover, the silver shells were also labeled with Rhodamine 6G. Fluorescence enhancement and reduced lifetime were also observed for silver-silica shell containing R6G. We found that by adjustment of probe concentration and silver shell thickness, a Ru(bpy)(3) (2+)-labeled particle could be 600 times brighter than an isolated Ru(bpy)(3) (2+) molecule. We expect labeled metal core-shell structures can become useful probes for high sensitivity and/or single particle assay.

UV-Vis spectroscopy and solvatochromism of the tyrosine kinase inhibitor AG-1478

NASA Astrophysics Data System (ADS)

Khattab, Muhammad; Wang, Feng; Clayton, Andrew H. A.

2016-07-01

The effect of twenty-one solvents on the UV-Vis spectrum of the tyrosine kinase inhibitor AG-1478 was investigated. The absorption spectrum in the range 300-360 nm consisted of two partially overlapping bands at approximately 340 nm and 330 nm. The higher energy absorption band was more sensitive to solvent and exhibited a peak position that varied from 327 nm to 336 nm, while the lower energy absorption band demonstrated a change in peak position from 340 nm to 346 nm in non-chlorinated solvents. The fluorescence spectrum of AG-1478 was particularly sensitive to solvent. The wavelength of peak intensity varied from 409 nm to 495 nm with the corresponding Stokes shift in the range of 64 nm to 155 nm (4536 cm- 1 to 9210 cm- 1). We used a number of methods to assess the relationship between spectroscopic properties and solvent properties. The detailed analysis revealed that for aprotic solvents, the peak position of the emission spectrum in wavenumber scale correlated with the polarity (dielectric constant or ET(30)) of the solvent. In protic solvents, a better correlation was observed between the hydrogen bonding power of the solvent and the position of the emission spectrum. Moreover, the fluorescence quantum yields were larger in aprotic solvents as compared to protic solvents. This analysis underscores the importance of polarity and hydrogen-bonding environment on the spectroscopic properties of AG-1478. These studies will assume relevance in understanding the interaction of AG-1478 in vitro and in vivo.

Fluorescence resonance energy transfer between green fluorescent protein and doxorubicin enabled by DNA nanotechnology.

PubMed

Heger, Zbynek; Kominkova, Marketa; Cernei, Natalia; Krejcova, Ludmila; Kopel, Pavel; Zitka, Ondrej; Adam, Vojtech; Kizek, Rene

2014-12-01

DNA nanotechnology is a rapidly growing research area, where DNA may be used for wide range of applications such as construction of nanodevices serving for large scale of diverse purposes. Likewise a panel of various purified fluorescent proteins is investigated for their ability to emit their typical fluorescence spectra under influence of particular excitation. Hence these proteins may form ideal donor molecules for assembly of fluorescence resonance emission transfer (FRET) constructions. To extend the application possibilities of fluorescent proteins, while using DNA nanotechnology, we developed nanoconstruction comprising green fluorescent protein (GFP) bound onto surface of surface active nanomaghemite and functionalized with gold nanoparticles. We took advantage of natural affinity between gold and thiol moieties, which were modified to bind DNA fragment. Finally we enclosed doxorubicin into fullerene cages. Doxorubicin intercalated in DNA fragment bound on the particles and thus we were able to connect these parts together. Because GFP behaved as a donor and doxorubicin as an acceptor using excitation wavelength for GFP (395 nm) in emission wavelength of doxorubicin (590 nm) FRET was observed. This nanoconstruction may serve as a double-labeled transporter of doxorubicin guided by force of external magnetic force owing to the presence of nanomaghemite. Further nanomaghemite offers the possibility of using this technology for thermotherapy. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fluorescence detection of esophageal neoplasia

NASA Astrophysics Data System (ADS)

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

2008-06-01

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

A look at some systemic properties of self-bioluminescent emission

NASA Astrophysics Data System (ADS)

Creath, Katherine

2008-08-01

Self-bioluminescent emission (SBE) is a type of biological chemiluminescence where photons are emitted as part of chemical reactions occurring during metabolic processes. This emission is also known as biophoton emission, ultraweak photon emission and ultraweak bioluminescence. This paper outlines research over the past century on some systemic properties of SBE as measured with biological detectors, photomultiplier detectors and ultra-sensitive imaging arrays. There is an apparent consensus in the literature that emission in the deep blue and ultraviolet (150-450nm) is related to DNA / RNA processes while emission in the red and near infrared (600-1000nm) is related to mitochondria and oxidative metabolisms involving reactive oxygen species, singlet oxygen and free radicals in plant, animal and human cells along with chlorophyll fluorescent decay in plants. Additionally, there are trends showing that healthy, unstressed and uninjured samples have less emission than samples that are unhealthy, stressed or injured. Mechanisms producing this emission can be narrowed down by isolating the wavelength region of interest and waiting for short-term fluorescence to decay leaving the ultraweak long-term metabolic emission. Examples of imaging this emission in healthy versus unhealthy, stressed versus unstressed, and injured versus uninjured plant parts are shown. Further discussion poses questions still to be answered related to properties such as coherence, photon statistics, and methodological means of isolating mechanisms.

A photostable near-infrared fluorescent tracker with pH-independent specificity to lysosomes for long time and multicolor imaging.

PubMed

Zhang, Xinfu; Wang, Chao; Han, Zhuo; Xiao, Yi

2014-12-10

A new boron-dipyrromethene-based lysosome tracker, Lyso-NIR, is facilely synthesized. Besides the intensive near-infrared (NIR) fluorescence and high photostability, Lyso-NIR shows the capability to stably localize in lysosomes, which is independent of the local pH. Lyso-NIR does not have the problematic alkalization effect suffered by the commonly used lysotrackers; thus, it shows ignorable cytotoxicity and slightly affects normal physiological functions of lysosomes. The above advantages of Lyso-NIR make it feasible to track lysosomes' dynamic changes in a relatively long time during the full cellular processes such as apoptosis, heavy metal stimulation, and endocytosis, as is demonstrated in this work. Moreover, Lyso-NIR's narrow NIR emission at 740 nm with a full width at half-maximum smaller than 50 nm makes it easy to avoid the crosstalk with the emissions from other common fluorescent probes, which strengthens Lyso-NIR's competitiveness as a standard lysosome tracker for multicolor bioimaging.

Fluorescence and phosphorescence of photomultiplier window materials under electron irradiation

NASA Technical Reports Server (NTRS)

Viehmann, W.; Eubanks, A. G.; Bredekamp, J. H.

1974-01-01

The fluorescence and phosphorescence of photomultiplier window materials under electron irradiation were investigated using a Sr-90/Y-90 beta emitter as the electron source. Spectral emission curves of UV grade, optical grade, and electron-irradiated samples of MGF2 and LiF, CaF2, BaF2, sapphire, fused silica, and UV transmitting glasses were obtained over the spectral range of 200 nm to 650 nm. Fluorescence yields, expressed as the number of counts in a solid angle of 2 pi steradian per 1MeV of incident electron energy deposited, were determined on these materials utilizing photomultiplier tubes with cesium telluride, bialkali, and trialkali (S-20) photocathodes, respectively.

Nitroolefin-based BODIPY as a novel water-soluble ratiometric fluorescent probe for detection of endogenous thiols

NASA Astrophysics Data System (ADS)

Kang, Jin; Huo, Fangjun; Chao, Jianbin; Yin, Caixia

2018-04-01

Small molecule biothiols, including cysteine (Cys), homocysteine (Hcy), and glutathione (GSH), play many crucial roles in physiological processes. In this work, we have prepared a nitroolefin-based BODIPY fluorescent probe with excellent water solubility for detection thiols, which displayed ratiometric fluorescent signal for thiols. Incorporation of a nitroolefin unit to the BODIPY dye would transform it into a strong Michael acceptor, which would be highly susceptible to sulfhydryl nucleophiles. This probe shows an obvious ratio change upon response with thiols, an increase of the emission at 517 nm along with a concomitant decrease of fluorescence peak at 573 nm. Moreover, these successes of intracellular imaging experiments in A549 cells indicated that this probe is suitable for imaging of ex-/endogenous thiols in living cells.

Host sensitized near-infrared emission in Nd3+ doped different alkaline-sodium-phosphate phosphors

NASA Astrophysics Data System (ADS)

Balakrishna, A.; Swart, H. C.; Kroon, R. E.; Ntwaeaborwa, O. M.

2018-04-01

Near-infrared (NIR) emitting phosphors of different alkaline based sodium-phosphate (MNa[PO4], where M = Mg, Ca, Sr and Ba were prepared by a conventional solution combustion method with fixed doping concentration of Nd3+ (1.0 mol%). The phosphors were characterized by powder X-ray diffraction, field emission scanning electron microscope, Fourier transform infrared spectroscopy, UV-vis spectroscopy and fluorescent spectrophotometry. The optical properties including reflectance, excitation and emission were investigated. The excitation spectra of the phosphors were characterized by a broadband extending from 450 to 900 nm. Upon excitation with a wavelength of 580 nm, the phosphor emits intensely infrared region at 872 nm, 1060 nm and 1325 nm which correspond to the 4F3/2 → 4I9/2, 4F3/2 → 4I11/2 and 4F3/2 → 4I13/2 transitions of Nd3+ ions and were found to vary for the different hosts. The strongest emission wavelength reaches 1060 nm. The most intense emission of Nd3+ was observed from Ca2+ incorporated host. The down conversion emissions of the material fall in the NIR region suggesting that the prepared phosphors have potential application in the development of photonic devices emitting in the NIR.

Recombination zone in white organic light emitting diodes with blue and orange emitting layers

NASA Astrophysics Data System (ADS)

Tsuboi, Taiju; Kishimoto, Tadashi; Wako, Kazuhiro; Matsuda, Kuniharu; Iguchi, Hirofumi

2012-10-01

White fluorescent OLED devices with a 10 nm thick blue-emitting layer and a 31 nm thick orange-emitting layer have been fabricated, where the blue-emitting layer is stacked on a hole transport layer. An interlayer was inserted between the two emitting layers. The thickness of the interlayer was changed among 0.3, 0.4, and 1.0 nm. White emission with CIE coordinates close to (0.33, 0.33) was observed from all the OLEDs. OLED with 0.3 nm thick interlayer gives the highest maximum luminous efficiency (11 cd/A), power efficiency (9 lm/W), and external quantum efficiency (5.02%). The external quantum efficiency becomes low with increasing the interlayer thickness from 0 nm to 1.0 nm. When the location of the blue- and orange-emitting layers is reversed, white emission was not obtained because of too weak blue emission. It is suggested that the electron-hole recombination zone decreases nearly exponentially with a distance from the hole transport layer.

Laser-induced fluorescence of phosphors for remote cryogenic thermometry

NASA Technical Reports Server (NTRS)

Beshears, D. L.; Capps, G. J.; Cates, M. R.; Simmons, C. M.; Schwenterly, S. W.

1990-01-01

Remote cryogenic temperature measurements can be made by inducing fluorescence in phosphors with temperature-dependent emissions and measuring the emission lifetimes. The thermographic phosphor technique can be used for making precision, noncontact, cryogenic-temperature measurements in electrically hostile environments, such as high dc electric or magnetic fields. The National Aeronautics and Space Administration is interested in using these thermographic phosphors for mapping hot spots on cryogenic tank walls. Europium-doped lanthanum oxysulfide (La2O2S:Eu) and magnesium fluorogermanate doped with manganese (Mg4FGeO6:Mn) are suitable for low-temperature surface thermometry. Several emission lines, excited by a 337-nm ultraviolet laser, provide fluorescence lifetimes having logarithmic dependence with temperature from 4 to above 125 K. A calibration curve for both La2O2S:Eu and Mg4FGeO6:Mn is presented, as well as emission spectra taken at room temperature and 11 K.

Dual color microscopic imagery of cells expressing the green fluorescent protein and a red-shifted variant.

PubMed

Yang, T T; Kain, S R; Kitts, P; Kondepudi, A; Yang, M M; Youvan, D C

1996-01-01

The green fluorescent protein (GFP) from the jellyfish, Aequorea victoria, has become a versatile reporter for monitoring gene expression and protein localization in a variety of cells and organisms. GFP emits bright green light (lambda max = 510 nm) when excited with ultraviolet (UV) or blue light (lambda max = 395 nm, minor peak at 470 nm). The chromophore in GFP is intrinsic to the primary structure of the protein, and fluorescence from GFP does not require additional gene products, substrates or other factors. GFP fluorescence is stable, species-independent and can be monitored noninvasively using the techniques of fluorescence microscopy and flow cytometry [Chalfie et al., Science 263 (1994) 802-805; Stearns, Curr. Biol. 5 (1995) 262-264]. The protein appears to undergo an autocatalytic reaction to create the fluorophore [Heim et al., Proc. Natl. Acad. Sci. USA 91 (1994) 12501-12504] in a process involving cyclization of a Tyr66 aa residue. Recently [Delagrave et al., Bio/Technology 13 (1995) 151-154], a combinatorial mutagenic strategy was targeted at aa 64 through 69, which spans the chromophore of A. victoria GFP, yielding a number of different mutants with red-shifted fluorescence excitation spectra. One of these, RSGFP4, retains the characteristic green emission spectra (lambda max = 505 nm), but has a single excitation peak (lambda max = 490 nm). The fluorescence properties of RSGFP4 are similar to those of another naturally occurring GFP from the sea pansy, Renilla reniformis [Ward and Cormier, Photobiochem. Photobiol. 27 (1978) 389-396]. In the present study, we demonstrate by fluorescence microscopy that selective excitation of A. victoria GFP and RSGFP4 allows for spectral separation of each fluorescent signal, and provides the means to image these signals independently in a mixed population of bacteria or mammalian cells.

Optical properties of flexible fluorescent films prepared by screen printing technology

NASA Astrophysics Data System (ADS)

Chen, Yan; Ke, Taiyan; Chen, Shuijin; He, Xin; Zhang, Mei; Li, Dong; Deng, Jinfeng; Zeng, Qingguang

2018-05-01

In this work, we prepared a fluorescent film comprised phosphors and silicone on flexible polyethylene terephthalate (PET) substrate using a screen printing technology. The effects of mesh number and weight ratio of phosphors to silicone on the optical properties of the flexible films were investigated. The results indicate that the emission intensity of the film increase as the mesh decreased from 400 to 200, but the film surface gradually becomes uneven. The fluorescent film with high emission intensity and smooth surface can be obtained when the weight ratio of phosphor to gel is 2:1, and mesh number is 300. The luminous efficiency of the fabricated LEDs combined the fluorescent films with 460 nm Ga(In)N chip module can reach 75 lm/W. The investigation indicates that the approach can be applied in the remote fluorescent film conversion and decreases the requirements of the particle size and the dispersion state of fluorescent materials.

Fluoride-driven 'turn on' ESPT in the binding with a novel benzimidazole-based sensor.

PubMed

Liu, Kai; Zhao, Xiaojun; Liu, Qingxiang; Huo, Jianzhong; Zhu, Bolin; Diao, Shihua

2015-01-01

A novel fluorescence sensor (BIP) bearing NH and OH subunits displayed a highly selective and sensitive recognition property for fluoride over other anions. Fluoride-driven ESPT, poorly used in anion recognition and sensing, was suggested to be responsible for the fluorescence enhancement with a blue shift of 35 nm in the emission spectrum.

Iron oxide nanozyme catalyzed synthesis of fluorescent polydopamine for light-up Zn2+ detection

NASA Astrophysics Data System (ADS)

Liu, Biwu; Han, Xiao; Liu, Juewen

2016-07-01

Fluorescent polydopamine (FPD) is an interesting material with excellent biocompatibility. However, its preparation is currently a lengthy and potentially dangerous process. We herein employ magnetic iron oxide (Fe3O4) nanoparticles as a peroxidase-mimicking nanozyme to produce FPD under mild conditions. Different from previous protocols using multiple steps with up to 6% (~2 M) H2O2, this preparation takes place in a single step with just 5 mM H2O2 at room temperature. The oxidized product shows excitation-wavelength-dependent emission peaks, similar to previous reports. The reaction kinetics, pH, temperature, and ionic strength are individually optimized. Among a diverse range of other nanomaterials tested, including Fe2O3, CeO2, CoO, Co3O4, NiO, TiO2, gold nanoparticles, and graphene oxide, Fe2O3 and graphene oxide yielded relatively weak emission, while the rest of the materials failed to produce FPD. The Fe3O4 nanoparticles retained ~90% catalytic activity even after ten cycles of synthesis. Finally, Zn2+ can enhance the fluorescence of FPD under 360 nm excitation but not under 480 nm excitation, leading to a sensitive light-up sensor with a detection limit of 60 nM Zn2+. Therefore, this work has demonstrated not only a novel use of nanozymes, but also an interesting application of FPD.Fluorescent polydopamine (FPD) is an interesting material with excellent biocompatibility. However, its preparation is currently a lengthy and potentially dangerous process. We herein employ magnetic iron oxide (Fe3O4) nanoparticles as a peroxidase-mimicking nanozyme to produce FPD under mild conditions. Different from previous protocols using multiple steps with up to 6% (~2 M) H2O2, this preparation takes place in a single step with just 5 mM H2O2 at room temperature. The oxidized product shows excitation-wavelength-dependent emission peaks, similar to previous reports. The reaction kinetics, pH, temperature, and ionic strength are individually optimized. Among a diverse range of other nanomaterials tested, including Fe2O3, CeO2, CoO, Co3O4, NiO, TiO2, gold nanoparticles, and graphene oxide, Fe2O3 and graphene oxide yielded relatively weak emission, while the rest of the materials failed to produce FPD. The Fe3O4 nanoparticles retained ~90% catalytic activity even after ten cycles of synthesis. Finally, Zn2+ can enhance the fluorescence of FPD under 360 nm excitation but not under 480 nm excitation, leading to a sensitive light-up sensor with a detection limit of 60 nM Zn2+. Therefore, this work has demonstrated not only a novel use of nanozymes, but also an interesting application of FPD. Electronic supplementary information (ESI) available: Methods, TEM, ζ-potential, and original fluorescence spectra. See DOI: 10.1039/c6nr02584f

Synthesis and Characterization of Water-Soluble Polythiophene Derivatives for Cell Imaging

NASA Astrophysics Data System (ADS)

Wang, Fengyan; Li, Meng; Wang, Bing; Zhang, Jiangyan; Cheng, Yongqiang; Liu, Libing; Lv, Fengting; Wang, Shu

2015-01-01

In this work, four water-soluble polythiophene derivatives (PT, PT-DDA, PT-ADA, and PT-ADA-PPR) with different pendant moieties were synthesized via oxidative copolymerization by FeCl3. By increasing the hydrophobic ability of side chain moieties, there is a gradually blue shift for the maximum absorption wavelength and red shift for the maximum emission wavelength, a reducing trend for fluorescence quantum yields, a growing trend for Stokes shift, and an increasing trend for the mean sizes in the order of PT, PT-ADA, and PT-DDA. All the synthesized polymers show low toxicity and good photostability and accumulate in the lysosomes of A549 cells. Furthermore, the introduction of porphyrin group to PT-ADA side chain (PT-ADA-PPR) broadens the absorption and emission ranges of PT-ADA. PT-ADA-PPR could be excited at two different excitation wavelengths (488 nm and 559 nm) and exhibits two emission pathways, and dual-color fluorescence images (orange and red) of PT-ADA-PPR accumulated in A549 cells are observed. Thus, PT-ADA-PPR could be used as an excellent dual-color fluorescent and lysosome-specific imaging material.

Fluorescence spectra of atmospheric aerosol at Adelphi, Maryland, USA: measurement and classification of single particles containing organic carbon

NASA Astrophysics Data System (ADS)

Pinnick, Ronald G.; Hill, Steven C.; Pan, Yong-Le; Chang, Richard K.

We measured laser-induced fluorescence spectra from individual supermicron-sized atmospheric particles drawn into our laboratory at Adelphi, MD, an urban site in the Washington, DC metroplex. A virtural impactor concentrator is used along with an aerodynamic-focusing-nozzle which forms, within an optical chamber, a focused aerosol jet where single aerosol particles can be interrogated on-the-fly with a pulsed 266-nm-wavelength laser. Sample rates are a few liter per minute, and are size dependent. Crossed-diode laser beams indicate when a particle is traversing the sample region and are used to trigger the UV laser to fire and the gated intensified CCD to record the fluorescence spectrum. Our breadboard fluorescence particle spectrometer measures particles in the 3-10 μm diameter size range. Typical trigger rates are a few per second. The usable spectral range is from about 295 to 605 nm. The majority of the particles have very weak fluorescence (on average 8 percent of particles have fluorescence signals above noise). The spectra were grouped using a heirarchical cluster analysis, with parameters chosen so that spectra typically cluster into 4-12 main categories. From the set of all cluster spectra we chose 8 template spectra for reanalyzing all the data. On average, 92 percent (81-94 percent) of the spectra were similar to these templates (using the same thresholds used for the cluster analysis). The major emission bands of the most commonly occurring spectra have peaks: near 460 nm (28 percent of fluorescent particles on average), a very broad hump, and may be humic acids or humic like substances; near 317 nm (on average 24 percent of fluorescent particles); near 321 and 460 nm (a double hump, 12 percent of fluorescent particles); and near 341 nm (8 percent of fluorescent particles). Some of the fluorescence in spectra peaking in the 317-341 nm range is probably from dicyclic aromatics and heterocyclics, including the amino acid tryptophan in biological particles such as bacteria and spores.

Enhancement of luminescence properties in Er3+ doped TeO2-Na2O-PbX (X=O and F) ternary glasses.

PubMed

Kumar, Kaushal; Rai, S B; Rai, D K

2007-04-01

An enhancement of luminescence properties in Er3+ doped ternary glasses is observed on the addition of PbO/PbF2. The infrared to visible upconversion emission bands are observed at 410, 525, 550 and 658 nm, due to the 2H9/2-->4I15/2, 2H11/2-->4I15/2, 4S3/2-->4I15/2, 4F9/2-->4I15/2 transitions respectively, on excitation with 797 nm laser line. A detailed study reveals that the 2H9/2-->4I15/2 transition arises due to three step upconversion process while other transitions arise due to two step absorption. On excitation with 532 nm radiation, ultraviolet and violet upconversion bands centered at 380, 404, 410 and 475 nm wavelengths are observed along with one photon luminescence bands at 525, 550, 658 and 843 nm wavelengths. These bands are found due to the 4G11/2-->4I15/2, 2P3/2-->4I13/2, 2H9/2-->4I15/2, 2P3/2-->4I11/2, 2H11/2-->4I15/2, 4S3/2-->4I15/2, 4F9/2-->4I15/2 and 4S3/2-->4I13/2 transitions, respectively. Though incorporation of PbO and PbF2 both enhances fluorescence intensities however, PbF2 content has an important influence on upconversion luminescence emission. The incorporation of PbF2 enhances the red emission (658 nm) intensity by 1.5 times and the violet emission (410 nm) intensity by 2.0 times. A concentration dependence study of fluorescence reveals the rapid increase in the red (4F9/2-->4I15/2) emission intensity relative to the green (4S3/2-->4I15/2) emission with increase in the Er3+ ion concentration. This behaviour has been explained in terms of an energy transfer by relaxation between excited ions.

Advances in Fluorescence Sensing Systems for the Remote Assessment of Nitrogen Supply in Field Corn

NASA Technical Reports Server (NTRS)

Corp, L. A.; Chappelle, E. W.; McMurtrey, J. E.; Daughtry, C. S. T.; Kim, M. S.

2000-01-01

The studies described herein were conducted to better define changes in fluorescence properties of leaves from field grown corn (Zea mays L.) as they relate to varying levels of nitrogen (N) fertilization. This research was directed toward: 1) providing a remote non-destructive sensing technique to aid in the determination of optimal rates of N fertilization in corn crops and, 2) defining parameters for further development of fluorescence instrumentation to be operated remotely at field canopy levels. Fluorescence imaging bands centered in the blue (450 nm), green (525 nm), red (680 nm), and far-red (740 nm) and ratios of these bands were compared with the following plant parameters: rates of photosynthesis, N:C ratio, pigment concentrations, and grain yields. Both the fluorescence and physiological measures exhibited similar curvilinear responses to N fertilization level while significant linear correlations were obtained among fluorescence bands and band ratios to certain physiological measures of plant productivity. The red / blue, red / green, far-red / blue, far-red /green fluorescence ratios are well suited for remote observation and provided high correlations to grain yield, LAI, N:C, and chlorophyll contents. The results from this investigation indicate that fluorescence technology could aid in the determination of N fertilization requirements for corn. This discussion will also address design concepts and preliminary field trials of a mobile field-based Laser Induced Fluorescence Imaging System (LIFIS) capable of simultaneously acquiring images of four fluorescence emission bands from areas of plant canopies equaling 1 sq m and greater without interference of ambient solar radiation.

A Cu2+-selective fluorescent chemosensor based on BODIPY with two pyridine ligands and logic gate

NASA Astrophysics Data System (ADS)

Huang, Liuqian; Zhang, Jing; Yu, Xiaoxiu; Ma, Yifan; Huang, Tianjiao; Shen, Xi; Qiu, Huayu; He, Xingxing; Yin, Shouchun

2015-06-01

A novel near-infrared fluorescent chemosensor based on BODIPY (Py-1) has been synthesized and characterized. Py-1 displays high selectivity and sensitivity for sensing Cu2+ over other metal ions in acetonitrile. Upon addition of Cu2+ ions, the maximum absorption band of Py-1 in CH3CN displays a red shift from 603 to 608 nm, which results in a visual color change from pink to blue. When Py-1 is excited at 600 nm in the presence of Cu2+, the fluorescent emission intensity of Py-1 at 617 nm is quenched over 86%. Notably, the complex of Py-1-Cu2+ can be restored with the introduction of EDTA or S2-. Consequently, an IMPLICATION logic gate at molecular level operating in fluorescence mode with Cu2+ and S2- as chemical inputs can be constructed. Finally, based on the reversible and reproducible system, a nanoscale sequential memory unit displaying "Writing-Reading-Erasing-Reading" functions can be integrated.

Denaturing of single electrospun fibrinogen fibers studied by deep ultraviolet fluorescence microscopy.

PubMed

Kim, Jeongyong; Song, Hugeun; Park, Inho; Carlisle, Christine R; Bonin, Keith; Guthold, Martin

2011-03-01

Deep ultraviolet (DUV) microscopy is a fluorescence microscopy technique to image unlabeled proteins via the native fluorescence of some of their amino acids. We constructed a DUV fluorescence microscope, capable of 280 nm wavelength excitation by modifying an inverted optical microscope. Moreover, we integrated a nanomanipulator-controlled micropipette into this instrument for precise delivery of picoliter amounts of fluid to selected regions of the sample. In proof-of-principle experiments, we used this instrument to study, in situ, the effect of a denaturing agent on the autofluorescence intensity of single, unlabeled, electrospun fibrinogen nanofibers. Autofluorescence emission from the nanofibers was excited at 280 nm and detected at ∼350 nm. A denaturant solution was discretely applied to small, select sections of the nanofibers and a clear local reduction in autofluorescence intensity was observed. This reduction is attributed to the dissolution of the fibers and the unfolding of proteins in the fibers. Copyright © 2010 Wiley-Liss, Inc.

Studies on fatty acid-binding proteins. The detection and quantification of the protein from rat liver by using a fluorescent fatty acid analogue.

PubMed Central

Wilkinson, T C; Wilton, D C

1986-01-01

Fatty acid-binding protein from rat liver is shown to bind the fluorescent fatty acid probe dansyl undecanoic acid. Binding is accompanied by a shift in the fluorescence emission maximum from 550 nm to 500 nm and a 60-fold fluorescence enhancement at 500 nm. These spectral properties have allowed the use of this probe to detect and quantify microgram amounts of liver fatty acid-binding protein during purification procedures. In conjunction with h.p.l.c. the method allows the rapid estimation of liver fatty acid-binding protein in biological samples. The validity of the method is demonstrated by measuring the concentration of fatty acid-binding protein in livers from control and hypolipidaemic-drug-treated rats. The dramatic diurnal rhythm previously reported for this protein [Dempsey (1984) Curr. Top. Cell. Regul. 24, 63-86] was not observed with this method. Images Fig. 1. PMID:3800946

Red emissive cross-linked chitosan and their nanoparticles for imaging the nucleoli of living cells.

PubMed

Wang, Ke; Yuan, Xun; Guo, Zhenpeng; Xu, Jiying; Chen, Yi

2014-02-15

Biocompatible glutaraldehyde-cross-linked chitosan with new red fluorescence were prepared for the first time and were shaped into nanoparticles via inverse-microemulsion method. They could luminesce at ca. 670 nm either as powders and nanoparticles or in real and gelling solutions or suspensions, having a lifetime of 1.353 ns and a quantum yield of 0.08 in solution or 0.01 in solid state. The new-formed pyridinium structures and the intramolecular charge transfer effect are considered to be responsible for the new red emission, which have been proved by FTIR, (13)C NMR, and some calculation using Gaussian 09, respectively. Strikingly, they are quite inert and anti-photobleaching, with only <3% loss of fluorescent intensity per minute in average under a continuous laser illumination at 633 nm and 50 μW. Especially, their nanoparticles (5.6 nm) could enter into the negative nucleoli of living HeLa cells with low cytotoxicity for high contrast imaging inspections. Copyright © 2013 Elsevier Ltd. All rights reserved.

The UV photochemistry of C2N2

NASA Technical Reports Server (NTRS)

Halpern, Joshua B.; Barts, Samuel A.

1989-01-01

The absorption, emission, and photodissociation yield spectra of C2N2 were measured in the 220 and 210 nm region near the 4(0)1 and 1(0)1 4(0)1 bands of the A 1 sigma + from the X 1 sigma + system. The emission spectrum showed very few lines which appeared in the absorption spectrum. Moreover, the emission had 660 ns lifetime and, at 210 nm a very large electronic emission quenching rate. Laser induced fluorescence was used to measure the relative yield of CN radicals as a function of photolysis wavelength. This spectrum seemed to follow the absorption spectrum below the dissociation threshold. Energy in the CN fragments appeared to be statistically distributed.

Using silicon-coated gold nanoparticles to enhance the fluorescence of CdTe quantum dot and improve the sensing ability of mercury (II)

NASA Astrophysics Data System (ADS)

Zhu, Jian; Chang, Hui; Li, Jian-Jun; Li, Xin; Zhao, Jun-Wu

2018-01-01

The effect of silicon-coated gold nanoparticles with different gold core diameter and silica shell thickness on the fluorescence emission of CdTe quantum dots (QDs) was investigated. For gold nanoparticles with a diameter of 15 nm, silica coating can only results in fluorescence recover of the bare gold nanoparticle-induced quenching of QDs. However, when the size of gold nanoparticle is increased to 60 nm, fluorescence enhancement of the QDs could be obtained by silica coating. Because of the isolation of the silica shell-reduced quenching effect and local electric field effect, the fluorescence of QDs gets intense firstly and then decreases. The maximum fluorescence enhancement takes place as the silica shell has a thickness of 30 nm. This enhanced fluorescence from silicon-coated gold nanoparticles is demonstrated for sensing of Hg2 +. Under optimal conditions, the enhanced fluorescence intensity decreases linearly with the concentration of Hg2 + ranging from 0 to 200 ng/mL. The limit of detection for Hg2 + is 1.25 ng/mL. Interference test and real samples detection indicate that the influence from other metal ions could be neglected, and the Hg2 + could be specifically detected.

Fluorescence multiplexing with time-resolved and spectral discrimination using a near-IR detector.

PubMed

Zhu, Li; Stryjewski, Wieslaw; Lassiter, Suzanne; Soper, Steven A

2003-05-15

We report on the design and performance of a two-color, time-resolved detector for the acquisition of both steady-state and time-resolved fluorescence data acquired in real time during the capillary gel electrophoresis separation of DNA sequencing fragments. The detector consisted of a pair of pulsed laser diodes operating at 680 and 780 nm. The diode heads were coupled directly to single-mode fibers, which were terminated into a single fiber mounted via a FC/PC connector to the detector body. The detector contained a dichroic filter, which directed the dual-laser beams to an objective. The objective focused the laser light into a capillary gel column and also collected the resulting fluorescence emission. The dual-color emission was transmitted through the dichroic and focused onto a multimode fiber (core diameter 50 microm), which carried the luminescence to a pair of single-photon avalanche diodes (SPADs). The emission was sorted spectrally using a second dichroic onto one of two SPADs and isolated using appropriate interference filters (710- or 810-nm channel). The dual-color detector demonstrated a time response of 450 and 510 ps (fwhm) for the 710- and 810-nm channels, respectively. The mass detection limits for two near-IR dye-labeled sequencing primers electrophoresed in a capillary gel column were found to be 7.1 x 10(-21) and 3.2 x 10(-20) mol (SNR = 3) for the 710- and 810-nm detector channels, respectively. In addition, no leakage of luminescence excited at 680 nm was observed in the 810-nm channel or 780-nm excited luminescence into the 710-nm channel. An M13mp18 template was sequenced in a single capillary gel column using a two-color, two-lifetime format. The read length was found to be 650 base pairs for the test template at a calling accuracy of 95.1% using a linear poly(dimethylacrylamide) (POP6) gel column, with the read length determined primarily by the electrophoretic resolution produced by the sieving gel.

Tuning of gold nanoclusters sensing applications with bovine serum albumin and bromelain for detection of Hg2+ ion and lambda-cyhalothrin via fluorescence turn-off and on mechanisms.

PubMed

Bhamore, Jigna R; Jha, Sanjay; Basu, Hirakendu; Singhal, Rakesh Kumar; Murthy, Z V P; Kailasa, Suresh Kumar

2018-04-01

Herein, fluorescent gold nanoclusters (Au NCs) were obtained by one-pot synthetic method using bovine serum albumin (BSA) and bromelain as templates. As-synthesized fluorescent Au NCs were stable and showed bright red fluorescence under UV lamp at 365 nm. The fluorescent Au NCs exhibit the emission intensity at 648 nm when excited at 498 nm. Various techniques were used such as spectroscopy (UV-visible, fluorescence, and Fourier-transform infrared), high-resolution transmission electron microscopy, and dynamic light scattering for the characterization of fluorescent Au NCs. The values of I 0 /I at 648 nm are proportional to the concentrations of Hg 2+ ion in the range from 0.00075 to 5.0 μM and of lambda-cyhalothrin in the range from 0.01 to 10 μM with detection limits of 0.0003 and 0.0075 μM for Hg 2+ ion and lambda-cyhalothrin, respectively. The practical application of the probe was successfully demonstrated by analyzing Hg 2+ ion and lambda-cyhalothrin in water samples. In addition, Au NCs used as probes for imaging of Simplicillium fungal cells. These results indicated that the as-synthesized Au NCs have proven to be promising fluorescent material for the sensing of Hg 2+ ion and lambda-cyhalothrin in environmental and for imaging of microorganism cells in biomedical applications.

Energy transfer dynamics of Er3+/Nd3+ embedded SiO2-Al2O3-Na2CO3-SrF2-CaF2 glasses for optical communications

NASA Astrophysics Data System (ADS)

Gelija, Devarajulu; Kadathala, Linganna; Borelli, Deva Prasad Raju

2018-04-01

The fluorescence and upconversion studies of Er3+ doped and Er3+/Nd3+ co-doped silicate based oxyfluoride glasses have been systematically analyzed. The broad band NIR emissions (830-1700 nm), includes optical bands like O, E, S, C and L were observed in the Er3+-Nd3+ co-doped glasses. The NIR emission intensity peaks centered at 876, 1057, 1329 and 1534 nm were observed for the Er3+-Nd3+ co-doped glasses. In the co-doped samples the strongest emission intensity at 1534 nm increased up to 0.5 mol % and then decreased to 3.0 mol % of Nd3+ ions under the excitation of 980 nm. The upconversion studies of the co-doped samples were recorded under the excitation of 980 and 808 nm and found the upconversion emission peaks centered at 524, 530, 547, 590 and 656 nm. The energy transfer processes between the relevant excitation levels of Er3+ and Nd3+ ions and energy transfer efficiency were discussed. The obtained results indicate that Nd3+ can be an efficient sensitizer for Er3+ to enhance upconversion emission at green laser transition for sensors and NIR emission at 1534 nm for optical communication applications.

Reflectivity Spectra for Commonly Used Reflectors

NASA Astrophysics Data System (ADS)

Janecek, Martin

2012-06-01

Monte Carlo simulations play an important role in developing and evaluating the performance of radiation detection systems. To accurately model a reflector in an optical Monte Carlo simulation, the reflector's spectral response has to be known. We have measured the reflection coefficient for many commonly used reflectors for wavelengths from 250 nm to 800 nm. The reflectors were also screened for fluorescence and angular distribution changes with wavelength. The reflectors examined in this work include several polytetrafluoroethylene (PTFE) reflectors, Spectralon, GORE diffuse reflector, titanium dioxide paint, magnesium oxide, nitrocellulose filter paper, Tyvek paper, Lumirror, Melinex, ESR films, and aluminum foil. All PTFE films exhibited decreasing reflectivity with longer wavelengths due to transmission. To achieve >;0.95 reflectivity in the 380 to 500 nm range, the PTFE films have to be at least 0.5 mm thick-nitrocellulose is a good alternative if a thin diffuse reflector is needed. Several of the reflectors have sharp declines in reflectivity below a cut-off wavelength, including TiO2 (420 nm), ESR film (395 nm), nitrocellulose (330 nm), Lumirror (325 nm), and Melinex (325 nm). PTFE-like reflectors were the only examined reflectors that had reflectivity above 0.90 for wavelengths below 300 nm. Lumirror, Melinex, and ESR film exhibited fluorescence. Lumirror and Melinex are excited by wavelengths between 320 and 420 nm and have their emission peaks located at 440 nm, while ESR film is excited by wavelengths below 400 nm and the emission peak is located at 430 nm. Lumirror and Melinex also exhibited changing angular distributions with wavelength.

Etching-dependent fluorescence quenching of Ag-dielectric-Au three-layered nanoshells: The effect of inner Ag nanosphere

NASA Astrophysics Data System (ADS)

Zhu, Jian; Xu, Zai-jie; Weng, Guo-jun; Zhao, Jing; Li, Jian-jun; Zhao, Jun-wu

2018-07-01

In this report, Ag-dielectric-Au three-layered nanoshells with controlled inner core size were synthesized. The fluorescence emission of the rhodamine 6G (R6G) could be quenched by the three-layered nanoshells distinctly. What's more, the fluorescence quenching efficiency could be further improved by tuning the etching of inner Ag nanosphere. The maximum fluorescence quenching efficiency is obtained when the separate layer just appears between the inner Ag core and the outer Au shell. Whereas the fluorescence quenching efficiency is weakened when no gaps take place around the inner Ag core or the separate layer is too thick and greater than 13 nm. The fluorescence quenching properties of the Ag-dielectric-Au three-layered nanoshells with different initial sizes of the Ag nanoparticles are also studied. The maximum fluorescence quenching efficiency is obtained when the three-layered nanoshells are synthesized based on the Ag nanoparticles with 60 nm, which is better than others two sizes (42 and 79 nm). Thus we believe that the size of initial Ag nanospheres also greatly affects the optimized fluorescence quenching efficiency. These results about fluorescence quenching properties of Ag-dielectric-Au three-layered nanoshells present a potential for design and fabrication of fluorescence nanosensors based on tuning the geometry of the inner core and the separate layer.

Synthesis, Characterization and Photophysical Properties of Pyridine-Carbazole Acrylonitrile Derivatives

PubMed Central

Pérez-Gutiérrez, Enrique; Percino, M. Judith; Chapela, Víctor M.; Cerón, Margarita; Maldonado, José Luis; Ramos-Ortiz, Gabriel

2011-01-01

We synthesized three novel highly fluorescent compounds, 2-(2’-pyridyl)-3-(N-ethyl-(3’-carbazolyl))acrylonitrile, 2-(3”-pyridyl)-3-(N-ethyl-(3’-carbazolyl))acrylonitrile, and 2-(4-pyridyl)-3-(N-ethyl-(3’-carbazolyl))acrylonitrile by Knoevenagel condensation. The first two were synthesized without solvent in the presence of piperidine as a catalyst; the third was synthesized without a catalyst and with N,N-dimethylformamide as a solvent. In solution, the molar absorption coefficients showed absorptions at 380, 378, and 396 nm, respectively; in solid state, absorptions were at 398, 390, and 442 nm, respectively. The fluorescence emission was at 540, 540 and 604 nm, respectively, the 2-(4-pyridyl)-3-(N-ethyl-(3’-carbazolyl))acrylonitrile showed a red shift in the emission of 64 nm compared to the other two compounds. The fluorescence quantum yield for the compounds in powder form showed values of 0.05, 0.14, and 0.006, respectively; compared with the value measured for the Alq3 reference, 2-(3”-pyridyl)-3-(N-ethyl-(3’-carbazolyl))acrylonitrile had a lightly higher value. The third harmonic generation measurement for 2-(2’-pyridyl)-3-(N-ethyl-(3’-carbazolyl))acrylonitrile yielded a χ(3) value of 5.5 × 10−12 esu, similar to that reported for commercial polymers. PMID:28880006

Two fluorescent wavelengths, 440(ex)/520(em) nm and 370(ex)/440(em) nm, reflect advanced glycation and oxidation end products in human skin without diabetes.

PubMed

Beisswenger, Paul J; Howell, Scott; Mackenzie, Todd; Corstjens, Hugo; Muizzuddin, Neelam; Matsui, Mary S

2012-03-01

Advanced glycation end products (AGEs) and oxidation products (OPs) play an important role in diabetes complications, aging, and damage from sun exposure. Measurement of skin autofluorescence (SAF) has been promoted as a noninvasive technique to measure skin AGEs, but the actual products quantified are uncertain. We have compared specific SAF measurements with analytically determined AGEs and oxidative biomarkers in skin collagen and determined if these measurements can be correlated with chronological aging and actinic exposure. SAF at four excitation (ex)/emission (em) intensities was measured on the upper inner arm ("sun protected") and dorsal forearm ("sun exposed") in 40 subjects without diabetes 20-60 years old. Skin collagen from the same sites was analyzed by liquid chromatography-tandem mass spectrometry for three AGEs-pentosidine, carboxymethyllysine (CML), and carboxyethyllysine (CEL)-and the OP methionine sulfoxide (MetSO). There was poor correlation of AGE-associated fluorescence spectra with AGEs and OP in collagen, with only pentosidine correlating with fluorescence at 370(ex)/440(em) nm. A little-studied SAF (440(ex)/520(em) nm), possibly reflecting elastin cross-links, correlated with all AGEs and OPs. Levels of CML, pentosidine, and MetSO, but not SAF, were significantly higher in sun-exposed skin. These AGEs and OPs, as well as SAF at 370(ex)/440(em) nm and 440(ex)/520(em) nm, increased with chronological aging. SAF measurements at 370(ex)/440(em) nm and 335(ex)/385(em) nm, except for pentosidine, which correlated with fluorescence at 370(ex)/440(em), correlate poorly with glycated and oxidatively modified protein in human skin and do not reflect actinic modification. A new fluorescence measurement (440(ex)/520(em) nm) appears to reflect AGEs and OPs in skin.

Environment-sensitive fluorophores with benzothiadiazole and benzoselenadiazole structures as candidate components of a fluorescent polymeric thermometer.

PubMed

Uchiyama, Seiichi; Kimura, Kohki; Gota, Chie; Okabe, Kohki; Kawamoto, Kyoko; Inada, Noriko; Yoshihara, Toshitada; Tobita, Seiji

2012-07-27

An environment-sensitive fluorophore can change its maximum emission wavelength (λ(em)), fluorescence quantum yield (Φ(f)), and fluorescence lifetime in response to the surrounding environment. We have developed two new intramolecular charge-transfer-type environment-sensitive fluorophores, DBThD-IA and DBSeD-IA, in which the oxygen atom of a well-established 2,1,3-benzoxadiazole environment-sensitive fluorophore, DBD-IA, has been replaced by a sulfur and selenium atom, respectively. DBThD-IA is highly fluorescent in n-hexane (Φ(f) =0.81, λ(em) =537 nm) with excitation at 449 nm, but is almost nonfluorescent in water (Φ(f) =0.037, λ(em) =616 nm), similarly to DBD-IA (Φ(f) =0.91, λ(em) =520 nm in n-hexane; Φ(f) =0.027, λ(em) =616 nm in water). A similar variation in fluorescence properties was also observed for DBSeD-IA (Φ(f) =0.24, λ(em) =591 nm in n-hexane; Φ(f) =0.0046, λ(em) =672 nm in water). An intensive study of the solvent effects on the fluorescence properties of these fluorophores revealed that both the polarity of the environment and hydrogen bonding with solvent molecules accelerate the nonradiative relaxation of the excited fluorophores. Time-resolved optoacoustic and phosphorescence measurements clarified that both intersystem crossing and internal conversion are involved in the nonradiative relaxation processes of DBThD-IA and DBSeD-IA. In addition, DBThD-IA exhibits a 10-fold higher photostability in aqueous solution than the original fluorophore DBD-IA, which allowed us to create a new robust molecular nanogel thermometer for intracellular thermometry. Copyright © 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Fluorescence Spectrum and Decay Measurement for Hsil VS Normal Cytology Differentiation in Liquid Pap Smear Supernatant

NASA Astrophysics Data System (ADS)

Vaitkuviene, A.; Gegzna, V.; Juodkazis, S.; Jursenas, S.; Miasojedovas, S.; Kurtinaitiene, R.; Rimiene, J.; Vaitkus, J.

2009-06-01

Cervical smear material contains endo and exocervical cells, mucus and inflammative, immune cells in cases of pathology. Just not destroyed keratinocytes lay on the glass for microscopy. Liquid cytology supernatant apart other diagnostics could be used for photodiagnostic. The spectroscopic parameters suitable for Normal and HSIL cytology groups supernatant differentiation are demonstrated. The dried liquid PAP supernatant fractions—sediment and liquid were investigated. Excitation and emission matrices (EEM), supernatant fluorescence decay measured under 280 nm diode short pulse excitation and fluorescence spectroscopy by excitation with 355 nm laser light were analyzed. The differences between Normal and HSIL groups were statistically proven in the certain spectral regions. Fluorescence decay peculiarities show spectral regions consisting of few fluorophores. Obtained results on fluorescence differences in Normal and HSIL groups' supernatant shows the potency of photodiagnosis application in cervical screening.

Fluorescence excitation and excited state intramolecular relaxation dynamics of jet-cooled methyl-2-hydroxy-3-naphthoate

NASA Astrophysics Data System (ADS)

McCarthy, Annemarie; Ruth, Albert A.

2013-11-01

Two distinct S0 → S1 fluorescence excitation spectra of methyl-2-hydroxy-3-napthoate (MHN23) have been obtained by monitoring fluorescence separately in the short (˜410 nm) and long (˜650 nm) wavelength emission bands. The short wavelength fluorescence is assigned to two MHN23 conformers which do not undergo excited state intramolecular proton transfer (ESIPT). Analysis of the 'long wavelength' fluorescence excitation spectrum, which arises from the proton transfer tautomer of MHN23 indicates an average lifetime of τ ⩾ 18 ± 2 fs for the initially excited states. Invoking the results of Catalan et al. [J. Phys. Chem. A, 1999, 103, 10921], who determined the N tautomer to decay predominantly via a fast non-radiative process, the limit of the rate of intramolecular excited proton transfer in MHN23 is calculated as, kpt ⩽ 1 × 1012 s-1.

A small molecular pH-dependent fluorescent probe for cancer cell imaging in living cell.

PubMed

Ma, Junbao; Li, Wenqi; Li, Juanjuan; Shi, Rongguang; Yin, Gui; Wang, Ruiyong

2018-05-15

A novel pH-dependent two-photon fluorescent molecular probe ABMP has been prepared based on the fluorophore of 2, 4, 6-trisubstituted pyridine. The probe has an absorption wavelength at 354 nm and corresponding emission wavelength at 475 nm with the working pH range from 2.20 to 7.00, especially owning a good liner response from pH = 2.40 to pH = 4.00. ABMP also has excellent reversibility, photostability and selectivity which promotes its ability in analytical application. The probe can be excited with a two-photon fluorescence microscopy and the fluorescence cell imaging indicated that the probe can distinguish Hela cancer cells out of normal cells with a two-photon fluorescence microscopy which suggested its potential application in tumor cell detection. Copyright © 2018 Elsevier B.V. All rights reserved.

A Passive Method for Detecting Vegetation Stress from Orbit: Chlorophyll Fluorescence Spectra from Fraunhofer Lines

NASA Technical Reports Server (NTRS)

Theisen, Arnold F.

2000-01-01

Solar-stimulated chlorophyll fluorescence measured with the Fraunhofer line depth method has correlated well with vegetation stress in previous studies. However, the instruments used in those studies were limited to a single solar absorption line (e.g. 656.3 nm), obviating the red/far-red ratio (R/FR) method. Optics and detector technology have reached the level whereby multiple, very narrow Fraunhofer lines are resolvable. Thirteen such lines span the visible spectrum in the red to far-red region where chlorophyll fluorescence occurs. Fluorescence intensities at the 13 Fraunhofer line wavelengths were used to model emission spectra. The source data were collected for summer and fall bean crops (Phaseolus vulgaris L.) subjected to various levels of nitrogen fertilization. The intensities were adjusted to account for Fraunhofer line depth and atmospheric transmittance. Multiple R/FR fluorescence ratios, calculated from the modeled fluorescence spectra, correlated strongly with leaf chlorophyll concentration and well with applied nitrogen. The ratio yielding the best correlation with chlorophyll utilized red fluorescence at the 694.5 nm Fraunhofer line and farred fluorescence at the 755.6 nm Fraunhofer line. Twenty R/FR ratios, each evaluated for the maximum differential between low and high (optimal) nitrogen treatments, ranked higher in some cases and lower in others, possibly related to the time of year the crops were grown and the stage of growth of the crops. Ratios with 728.9 nm and 738.9 nm in the denominator consistently ranked in the lowest and next lowest quartile, respectively. Ratios of the 656.3 nm Fraunhofer line and the 755.6 nm line consistently ranked highest for the summer crop. Ratios with 755.6 nm in the denominator ranked in the upper quartile for 10 out of 12 measurement dates. Differences in ratio ranking indicate that physiological conditions may be estimated using selected ratios of Fraunhofer lines within the context of R/FR analysis. A passive instrument designed to monitor R/FR chlorophyll fluorescence (i.e. vegetation stress) from orbit could be built today.

Laser-induced fluorescence ratios of Cajanus cajan L. under the stress of cadmium and its correlation with pigment content and pigment ratios.

PubMed

Maurya, Renu; Gopal, R

2008-04-01

Laser-induced fluorescence spectra were used to characterize the effect of cadmium on the pigment status of the leaves of Cajanus cajan L. Laser-induced fluorescence spectra of untreated as well as cadmium treated (0.01 mM, 0.10 mM, and 1.00 mM) Cajanus cajan L. were recorded using the 355 nm line of a Nd:YAG laser as the excitation source and a monochromator with an intensified charge-coupled device as a detector in the region 400-800 nm. The fluorescence intensity ratios (FIR) of control as well as treated Cajanus cajan L. have been calculated by evaluating curve fitted parameters using a Gaussian spectral function. In addition, some growth parameters, such as photosynthetic pigment content, were also measured. The 355 nm line of the laser-light-excited leaves not only showed a fluorescence emission in the red spectral region (650-800 nm), but also in the blue-green region (400-570 nm). The chlorophyll FIR F690/F740 strongly correlated with the photosynthetic pigment content (total chlorophyll and carotenoids) and its ratio. Consequently, a correlation was also seen between the ratio of the blue-green fluorescence F470/F540 and the photosynthetic pigment content. The results indicated that the plants treated with 0.01 mM of cadmium exhibited better growth, while higher concentrations of cadmium were hazardous for Cajanus cajan L.

Green fluorescent protein as a reporter of gene expression and protein localization.

PubMed

Kain, S R; Adams, M; Kondepudi, A; Yang, T T; Ward, W W; Kitts, P

1995-10-01

The green fluorescent protein (GFP) from the jellyfish Aequorea victoria is rapidly becoming an important reporter molecule for monitoring gene expression and protein localization in vivo, in situ and in real time. GFP emits bright green light (lambda max = 509 nm) when excited with UV or blue light (lambda max = 395 nm, minor peak at 470 nm). The fluorescence excitation and emission spectra of GFP are similar to those of fluorescein, and the conditions used to visualize this fluorophore are also suitable for GFP. Unlike other bioluminescent reporters, the chromophore in GFP is intrinsic to the primary structure of the protein, and GFP fluorescence does not require a substrate or cofactor. GFP fluorescence is stable, species-independent and can be monitored non-invasively in living cells and, in the case of transparent organisms, whole animals. Here we demonstrate GFP fluorescence in bacterial and mammalian cells and introduce our Living Colors line of GFP reporter vectors, GFP protein and anti-GFP antiserum. The reporter vectors for GFP include a promoterless GFP vector for monitoring the expression of cloned promoters/enhancers in mammalian cells and a series of six vectors for creating fusion protein to either the N or C terminus of GFP.

Pigment organization effects on energy transfer and Chl a emission imaged in the diatoms C. meneghiniana and P. tricornutum in vivo: a confocal laser scanning fluorescence (CLSF) microscopy and spectroscopy study.

PubMed

Premvardhan, Lavanya; Réfrégiers, Matthieu; Büchel, Claudia

2013-09-26

The (auto)fluorescence from three diatom strains, Cyclotella meneghiniana (Cm), Phaeodactylum tricornutum 1a (Pt1a), and Phaeodactylum UTex (PtUTex), has been imaged in vivo to submicrometer resolution using confocal laser scanning fluorescence (CLSF) microscopy. The diatoms are excited at 473 and 532 nm, energy primarily absorbed by the carotenoid fucoxanthin (Fx) found within the fucoxanthin chlorophyll a/c proteins (FCPs). On the basis of the fluorescence spectra measured in each image voxel, we obtain information about the spatial and energetic distribution of the terminal Chl a emitters, localized in the FCPs and the reaction centers of the PSII protein complexes, and the nature and location of the primary absorbers that are linked to these emitters; 532 nm excites the highly efficient Fx(red) light harvesters, and lesser amounts of Fx(green)s, that are enriched in some FCPs and preferentially transfer energy to PSII, compared to 473 nm, which excites almost equal amounts of all three previously identified sets of Fx--Fx(red), Fx(green) and Fx(blue)--as well as Chl c. The heterogeneous Chl a emission observed from the (C)LSF images indicates that the different Fx's serve different final emitters in P. tricornutum and suggest, at least in C. meneghiniana , a localization of FCPs with relatively greater Fx(red) content at the chloroplast edges, but with overall higher FCP concentration in the interior of the plastid. To better understand our results, the concentration-dependent ensemble-averaged diatom solution spectra are compared to the (auto)fluorescence spectra of individual diatoms, which indicate that pigment packing effects at an intracellular level do affect the diatoms' spectral properties, in particular, concerning a 710 nm emission band apparent under stress conditions. A species-specific response of the spectral signature to the incident light is also discussed in terms of the presence of a silica shell in Cm but not in Pt1a nor PtUTex.

Near-infrared fluorescence imaging using organic dye nanoparticles.

PubMed

Yu, Jia; Zhang, Xiujuan; Hao, Xiaojun; Zhang, Xiaohong; Zhou, Mengjiao; Lee, Chun-Sing; Chen, Xianfeng

2014-03-01

Near-infrared (NIR) fluorescence imaging in the 700-1000 nm wavelength range has been very attractive for early detection of cancers. Conventional NIR dyes often suffer from limitation of low brightness due to self-quenching, insufficient photo- and bioenvironmental stability, and small Stokes shift. Herein, we present a strategy of using small-molecule organic dye nanoparticles (ONPs) to encapsulate NIR dyes to enable efficient fluorescence resonance energy transfer to obtain NIR probes with remarkably enhanced performance for in vitro and in vivo imaging. In our design, host ONPs are used as not only carriers to trap and stabilize NIR dyes, but also light-harvesting agent to transfer energy to NIR dyes to enhance their brightness. In comparison with pure NIR dyes, our organic dye nanoparticles possess almost 50-fold increased brightness, large Stokes shifts (∼250 nm) and dramatically enhanced photostability. With surface modification, these NIR-emissive organic nanoparticles have water-dispersity and size- and fluorescence- stability over pH values from 2 to 10 for almost 60 days. With these superior advantages, these NIR-emissive organic nanoparticles can be used for highly efficient folic-acid aided specific targeting in vivo and ex vivo cellular imaging. Finally, during in vivo imaging, the nanoparticles show negligible toxicity. Overall, the results clearly display a potential application of using the NIR-emissive organic nanoparticles for in vitro and in vivo imaging. Copyright © 2014 Elsevier Ltd. All rights reserved.

Photoswitchable red fluorescent protein with a large Stokes shift.

PubMed

Piatkevich, Kiryl D; English, Brian P; Malashkevich, Vladimir N; Xiao, Hui; Almo, Steven C; Singer, Robert H; Verkhusha, Vladislav V

2014-10-23

A subclass of fluorescent proteins (FPs), large Stokes shift (LSS) FP, are characterized by increased spread between excitation and emission maxima. We report a photoswitchable variant of a red FP with an LSS, PSLSSmKate, which initially exhibits excitation and emission at 445 and 622 nm, but violet irradiation photoswitches PSLSSmKate into a common red form with excitation and emission at 573 and 621 nm. We characterize spectral, photophysical, and biochemical properties of PSLSSmKate in vitro and in mammalian cells and determine its crystal structure in the LSS form. Mass spectrometry, mutagenesis, and spectroscopy of PSLSSmKate allow us to propose molecular mechanisms for the LSS, pH dependence, and light-induced chromophore transformation. We demonstrate the applicability of PSLSSmKate to superresolution photoactivated localization microscopy and protein dynamics in live cells. Given its promising properties, we expect that PSLSSmKate-like phenotype will be further used for photoactivatable imaging and tracking multiple populations of intracellular objects.

A simple turn on fluorescent sensor for the selective detection of thiamine using coconut water derived luminescent carbon dots.

PubMed

Purbia, Rahul; Paria, Santanu

2016-05-15

In this study microwave-assisted hydrothermal method was used to prepare highly luminescent carbon dots (1-6 nm size) within a minute from tender coconut (Cocos nucifera) water. The synthesized carbon dots (C-dots) exhibit emission of blue and green lights while excited at 390 and 450 nm wavelengths, respectively. As an application, these C-dots were tested for a simple "turn on" fluorescent sensor for rapid detection of thiamine (vitamin B1). The detection of thiamine in human body is very important to prevent various diseases such as beriberi, neurological disorders, optic neuropathy, etc. The fluorescence emission intensity of C-dots quenches after addition of Cu(2+) ion and then again increases selectively (turn on) after the addition of thiamine. The fluorescence emission intensity enhancement of Cu(2+) ion modified C-dots in the presence of thiamine exhibits a linear relationship within the thiamine concentration range of 10-50 μM. The limit of detection was found to be 280 nM from this study. The selectivity of the detection was also tested in the presence of different organic molecules and inorganic ions (Ca(2+), Mg(2+), Na(+), K(+), Cl(-), SO4(2-), and NO3(-)) which are present in blood serum and urine and found to be almost no interference in the detection. Finally, to see the applicability in real samples a commercial vitamin capsule was tested and found less than 3% error in the detected concentration. The C-dots were also used for bioimaging of fungus and the results show they are also suitable for this application too. Copyright © 2015 Elsevier B.V. All rights reserved.

Design of mitochondria-targeted colorimetric and ratiometric fluorescent probes for rapid detection of SO2 derivatives in living cells

NASA Astrophysics Data System (ADS)

Yang, Yutao; Zhou, Tingting; Bai, Bozan; Yin, Caixia; Xu, Wenzhi; Li, Wei

2018-05-01

Two mitochondria-targeted colorimetric and ratiometric fluorescent probes for SO2 derivatives were constructed based on the SO2 derivatives-triggered Michael addition reaction. The probes exhibit high specificity toward HSO3-/SO32- by interrupting their conjugation system resulting in a large ratiometric blue shift of 46-121 nm in their emission spectrum. The two well-resolved emission bands can ensure accurate detection of HSO3-. The detection limits were calculated to be 1.09 and 1.35 μM. Importantly, probe 1 and probe 2 were successfully used to fluorescence ratiometric imaging of endogenous HSO3- in BT-474 cells.

Measuring evaporation rates of laser-trapped droplets by use of fluorescent morphology-dependent resonances.

PubMed

Pastel, R; Struthers, A

2001-05-20

Morphology-dependent resonances (MDRs) are used to measure accurately the evaporation rates of laser-trapped 1- to 2-mum droplets of ethylene glycol. Droplets containing 3 x 10(-5) M Rhodamine-590 laser dye are optically trapped in a 20-mum hollow fiber by two counterpropagating 150-mW, 800-nm laser beams. A weaker 532-nm laser excites the dye, and fluorescence emission is observed near 560 nm as the droplet evaporates. A complete series of first-order TE and TM MDRs dominates the fluorescent output. MDR mode identification sizes the droplets and provides accurate evaporation rates. We verify the automated MDR mode identification by counting fringes in a videotape of the experiment. The longitudinal spring constant of the trap, measured by analysis of the videotaped motion of droplets perturbed from the trap center, provides independent verification of the laser's intensity within the trap.

Measuring Evaporation Rates of Laser-Trapped Droplets by Use of Fluorescent Morphology-Dependent Resonances

NASA Astrophysics Data System (ADS)

Pastel, Robert; Struthers, Allan

2001-05-01

Morphology-dependent resonances (MDRs) are used to measure accurately the evaporation rates of laser-trapped 1- to 2- m droplets of ethylene glycol. Droplets containing 3 x10-5 M Rhodamine-590 laser dye are optically trapped in a 20- m hollow fiber by two counterpropagating 150-mW, 800-nm laser beams. A weaker 532-nm laser excites the dye, and fluorescence emission is observed near 560 nm as the droplet evaporates. A complete series of first-order TE and TM MDRs dominates the fluorescent output. MDR mode identification sizes the droplets and provides accurate evaporation rates. We verify the automated MDR mode identification by counting fringes in a videotape of the experiment. The longitudinal spring constant of the trap, measured by analysis of the videotaped motion of droplets perturbed from the trap center, provides independent verification of the laser s intensity within the trap.

Dayglow emissions of the O2 Herzberg bands and the Rayleigh backscattered spectrum of the earth

NASA Technical Reports Server (NTRS)

Frederick, J. E.; Abrams, R. B.

1982-01-01

It is pointed out that numerous fluorescent emissions from the Herzberg bands of molecular oxygen lie in the spectral region 242-300 nm. This coincides with the wavelength range used by orbiting spectrometers that observe the Rayleigh backscattered spectrum of the earth for the purpose of monitoring the vertical distribution of stratospheric ozone. Model calculations suggest that Herzberg band emissions in the dayglow could provide significant contamination of the ozone measurements if the quenching rate of O2(A3Sigma) is sufficiently small. It is noted that this is especially true near 255 nm, where the most intense fluorescent emissions relative to the Rayleigh scattered signal are located and where past satellite measurements have shown a persistent excess radiance above that expected for a pure ozone absorbing and molecular scattering atmosphere. Very small quenching rates, however, are adequate to reduce the dayglow emission to negligible levels. Noting that available laboratory data have not definitely established the quenching on the rate of O2(A3Sigma) as a function of vibration level, it is emphasized that such information is required before the Herzberg band contributions can be evaluated with confidence.

Cryogenic Tm:YAP microchip laser

NASA Astrophysics Data System (ADS)

Hubka, Zbyněk.; Å ulc, Jan; Jelínková, Helena; Nejezchleb, Karel; Å koda, Václav

2016-04-01

The spectral characteristics of laser active media, and thus those of the laser output, are temperature dependent. Specifically, in almost every crystal host, cooling to low temperatures leads to better heat removal, a higher efficiency and output power, and a reduced lasing threshold. Tm-ion doped lasers have an emission wavelength around 2 μm and are important in medicine for soft tissue cutting and hemostasis, as well as in LIDAR or atmosphere sensing technology. This paper presents the performance-temperature dependency of a 4 at. % doped Tm:YAP microchip. During the experiment the Tm:YAP crystal was placed inside an evacuated liquid nitrogen cryostat on a cooling finger. As its temperature was varied from 80 K to 340 K, changes were observed in the absorption spectrum, ranging from 750 nm to 2000 nm and in the fluorescence spectrum from 1600 nm to 2050 nm. Fluorescence lifetime was seen to rise and fall with decreasing temperature. The laser was pumped by a 792 nm laser diode and at 80 K the maximum output peak power of the laser was 4.6 W with 23 % slope efficiency and 0.6 W threshold, compared to 2.4 W output peak power, 13 % slope efficiency and 3.3 W threshold when at 340 K. The laser emission wavelength changed from 1883 nm to 1993 nm for 80 K and 300 K, respectively.

First derivative emission spectrofluorimetric method for the determination of LCZ696, a newly approved FDA supramolecular complex of valsartan and sacubitril in tablets.

PubMed

Ragab, Marwa A A; Galal, Shereen M; Korany, Mohamed A; Ahmed, Aya R

2017-12-01

LCZ696 (sacubitril/valsartan, Entresto™) is a therapy lately approved by United States Food and Drug Administration (US FDA) as a heart failure therapy. It is claimed to decrease the mortality rate and hospitalization for patients with chronic heart failure. This study is considered as the first report to investigate the fluorimetric behavior of sacubitril in addition to pursuing all the different conditions that may affect its fluorescence. Various conditions were studied, for example studying the effects of organized media, solvents and pH, which may affect the fluorescence behavior of sacubitril. For the simultaneous determination of the newly approved supramolecular complex of valsartan (VAL) and sacubitril (SAC) in their tablets, a sensitive and simple first derivative spectrofluorimetric method was developed. The method involved the measurement of native fluorescence at 416 nm and 314 nm (λ ex 249 nm) for VAL and SAC, respectively. The first (D1) derivative technique was applied to the emission data to resolve a partial overlap that appeared in their emission spectra. The proposed method was successfully applied for the assay of the two drugs in their supramolecular complex LCZ696 with no interference from common pharmaceutical additives. International Conference on Harmonisation of Technical Requirements for Registration of Pharmaceuticals for Human Use (ICH) guidelines were followed in order to validate the proposed method. Copyright © 2017 John Wiley & Sons, Ltd.

Fine tuning the emission wavelengths of the 7-hydroxy-1-indanone based nano-structure dyes: Near-infrared (NIR) dual emission generation with large stokes shifts

NASA Astrophysics Data System (ADS)

Roohi, Hossein; Alizadeh, Parvaneh

2018-05-01

Near-infrared (NIR) fluorescent dyes have recently gained special attention due to their applications to use as molecular probes for imaging of biological targets and sensitive determination. In this study, photophysical properties of the 7-hydroxy-1-indanone based fluorophors A1, A2, A3, B1, B2 and 3R-B2 (R = CF3, NH2, NO2 and OMe) in the gas and three solution phases were probed using TD-DFT method at PBE0/6-311++G(d,p) and M06-2X/6-311++G(d,p) levels of theory. In addition to structural and photophysical properties as well as ESIPT mechanism of all mentioned molecules, the FC and relaxed potential energy surfaces of B2 and 3R-B2 (R = CF3 and NH2) molecules were explored in gas phase and acetonitrile, cyclohexane and water solvents. It is predicted that the A1, A3 and 3R-B2 chromophores afford normal (615-670 nm) and NIR fluorescence emissions (770-940 nm; biological window) with the large Stokes shifts of >160 and >300 nm, respectively. A good aggrement was found between theoretical and experimental results. In sum, these new types of dyes may render the new approaches for the development of the most efficient NIR fluorescent probes for enhanced image contrast and optimal apparent brightness in biological applications.

Vacuum-UV fluorescence spectroscopy of PF3 in the range 9-20 eV

NASA Astrophysics Data System (ADS)

Biehl, H.; Boyle, K. J.; Seccombe, D. P.; Tuckett, R. P.; Baumgärtel, H.; Jochims, H. W.

1998-01-01

The vacuum-UV and visible spectroscopy of PF3 using fluorescence excitation and dispersed emission techniques is reported. The fluorescence excitation spectrum has been recorded following photoexcitation with monochromatized synchrotron radiation from the Daresbury, UK source in the energy range 9-20 eV with an average resolution of ˜0.015 eV. Transitions to the three lowest-energy bands in the Rydberg spectra show resolved vibrational structure, they are assigned to transitions to the (8a1)-1 4p, 5p, and 6p Rydberg states of PF3, and fluorescence is due to valence transitions in the PF2 radical. From a Franck-Condon analysis of the vibrational structure, it is shown that the FPF bond angle in PF3 increases by ˜14±1° upon photoexcitation. The use of optical filters shows that at least three excited electronic states of PF2 are responsible for the induced emission. Dispersed emission spectra in the UV/visible region have been recorded with an optical resolution of 8 nm at the BESSY 1, Germany synchrotron source at the energies of all the peaks in the excitation spectrum. Four different decay channels are observed: (a) PF2 Ã 2A1-X˜2B1 fluorescence in the wide range 320-550 nm for photon energies around 9.8 eV, (b) PF2 ÖX˜, and B˜ 2B2-X˜ 2B1 fluorescence at ˜300 nm for photon energies around 11.0 eV, (c) PF2 C˜ 2A1-X˜ 2B1 and Ẽ 2B1 (2Π)-Ã 2A1 fluorescence at ˜222 and 325 nm, respectively, for photon energies around 14.4 eV, and (d) PF A 3Π-X 3Σ- fluorescence between 300-380 nm for photon energies around 16.1 eV. These assignments are confirmed by action spectra in which the excitation energy of the vacuum-UV radiation is scanned with detection of the fluorescence at a fixed, dispersive wavelength. Using the single-bunch mode of the BESSY 1 source, we have attempted to measure the lifetimes of the emitting states, but the timing profile of the source imposes an upper limit on lifetimes that can be measured of ˜500 ns. We have therefore only been able to measure values for the bent C˜ 2A1 and linear Ẽ 2B1 (2Π) states of PF2 of 14.7 and 7.9 ns, respectively; the lifetimes of the other emitters are too long to measure by this method. Our assignments in PF2 are heavily dependent on recent ab initio calculations on the geometries and energies of the valence electronic states of this species. Our knowledge on the electronic spectroscopy of this free radical is reviewed.

Red shift in the spectrum of a chlorophyll species is essential for the drought-induced dissipation of excess light energy in a poikilohydric moss, Bryum argenteum.

PubMed

Shibata, Yutaka; Mohamed, Ahmed; Taniyama, Koichiro; Kanatani, Kentaro; Kosugi, Makiko; Fukumura, Hiroshi

2018-05-01

Some mosses are extremely tolerant of drought stress. Their high drought tolerance relies on their ability to effectively dissipate absorbed light energy to heat under dry conditions. The energy dissipation mechanism in a drought-tolerant moss, Bryum argenteum, has been investigated using low-temperature picosecond time-resolved fluorescence spectroscopy. The results are compared between moss thalli samples harvested in Antarctica and in Japan. Both samples show almost the same quenching properties, suggesting an identical drought tolerance mechanism for the same species with two completely different habitats. A global target analysis was applied to a large set of data on the fluorescence-quenching dynamics for the 430-nm (chlorophyll-a selective) and 460-nm (chlorophyll-b and carotenoid selective) excitations in the temperature region from 5 to 77 K. This analysis strongly suggested that the quencher is formed in the major peripheral antenna of photosystem II, whose emission spectrum is significantly broadened and red-shifted in its quenched form. Two emission components at around 717 and 725 nm were assigned to photosystem I (PS I). The former component at around 717 nm is mildly quenched and probably bound to the PS I core complex, while the latter at around 725 nm is probably bound to the light-harvesting complex. The dehydration treatment caused a blue shift of the PS I emission peak via reduction of the exciton energy flow to the pigment responsible for the 725 nm band.

[Verification of ballast water exchange for international ships anchored in Xiamen Port by CDOM tracer].

PubMed

Li, Chao; Zhang, Yan-po; Guo, Wei-dong; Zhu, Yue; Xu, Jing; Deng, Xun

2010-09-01

Fluorescence excitation-emission matrix (EEM) and absorption spectroscopy were applied to study the optical properties of 29 CDOM samples collected from different ballast tanks of nine international route vessels anchored in Xiamen Port between October 2007 and April 2008. The purpose was to examine the feasibility of these spectral properties as a tracer to verify if these vessels follow the mid-ocean ballast water exchange (BWE) regulation. Using parallel factor analysis, four fluorescent components were identified, including two humic-like components (C1: 245, 300/386 nm; C2: 250, 345/458 nm) and two protein-like components (C3: 220, 275/306 nm; C4: 235, 290/345 nm), of which C2 component was the suitable fluorescence verification indicator. The vertical distribution of all fluorescent components in ballast tank was nearly similar indicating that profile-mixing sampling was preferable. Combined use of C2 component, spectral slope ratio (SR) of absorption spectroscopy and salinity may provide reasonable verification if BWE carried out by these nine ships. The results suggested that the combined use of multiple parameters (fluorescence, absorption and salinity) would be much reliable to determine the origin of ballast water, and to provide the technical guarantee for fast examination of ballast water exchange in Chinese ports.

Optimal parameters for near infrared fluorescence imaging of amyloid plaques in Alzheimer’s disease mouse models

PubMed Central

Raymond, S B; Kumar, A T N; Boas, D A; Bacskai, B J

2012-01-01

Amyloid-β plaques are an Alzheimer’s disease biomarker which present unique challenges for near-infrared fluorescence tomography because of size (<50 μm diameter) and distribution. We used high-resolution simulations of fluorescence in a digital Alzheimer’s disease mouse model to investigate the optimal fluorophore and imaging parameters for near-infrared fluorescence tomography of amyloid plaques. Fluorescence was simulated for amyloid-targeted probes with emission at 630 and 800 nm, plaque-to-background ratios from 1–1000, amyloid burden from 0–10%, and for transmission and reflection measurement geometries. Fluorophores with high plaque-to-background contrast ratios and 800 nm emission performed significantly better than current amyloid imaging probes. We tested idealized fluorophores in transmission and full-angle tomographic measurement schemes (900 source–detector pairs), with and without anatomical priors. Transmission reconstructions demonstrated strong linear correlation with increasing amyloid burden, but underestimated fluorescence yield and suffered from localization artifacts. Full-angle measurements did not improve upon the transmission reconstruction qualitatively or in semi-quantitative measures of accuracy; anatomical and initial-value priors did improve reconstruction localization and accuracy for both transmission and full-angle schemes. Region-based reconstructions, in which the unknowns were reduced to a few distinct anatomical regions, produced highly accurate yield estimates for cortex, hippocampus and brain regions, even with a reduced number of measurements (144 source–detector pairs). PMID:19794239

Optical redox ratio differentiates early tissue transformations in DMBA-induced hamster oral carcinogenesis based on autofluorescence spectroscopy coupled with multivariate analysis

NASA Astrophysics Data System (ADS)

Sethupathi, R.; Gurushankar, K.; Krishnakumar, N.

2016-11-01

Fluorescence intensity measurements have the potential to facilitate the diagnoses of many pathological conditions. The changes in fluorescence intensity may be influenced by factors such as tissue architectures, endogenous fluorophores, cellular metabolism and light penetration depth in tissue. Two of the most diagnostically important endogenous fluorophores are reduced nicotinamide dinucleotide (NADH) and flavin adenine dinucleotide (FAD), which can be used to monitor dramatic metabolic changes in cells and tissues. The goal of this study is to investigate changes in the endogenous fluorophore emission and to quantify metabolic changes in the redox state of various tissue transformation conditions with respect to control tissues in dimethyl benz[a] anthracene (DMBA)-induced hamster oral carcinogenesis for measuring emission spectrum at 320 nm excitation. In the present study, collagen, NADH and FAD emission of well-differentiated squamous cell carcinoma (WDSCC) showed decreased intensity at ~385 nm, ~450 nm and ~520 nm compared to hyperplasia, dysplasia and control tissues. Furthermore, a significant decrease in the optical redox ratio is observed in WDSCC tissues, which indicates an increased metabolic activity compared to the control tissues. Moreover, the principal component linear discriminant analysis (PC-LDA) algorithm together with the leave-one-out cross-validation (LOOCV) method yield an overall diagnostic sensitivity of 77.7% and a specificity of 88.8% in the classification of control, hyperplasia, dysplasia and WDSCC tissues, respectively. The results from this study demonstrated that fluorescence-based tissue analysis combined with PC-LDA has tremendous potential for the effective discrimination of control from neoplastic tissues; furthermore it also detects early neoplastic changes prior to definite morphologic alteration.

Spectral and time-resolved studies on ocular structures

NASA Astrophysics Data System (ADS)

Schweitzer, D.; Jentsch, S.; Schenke, S.; Hammer, M.; Biskup, C.; Gaillard, E.

2007-07-01

Measurements of endogeous fluorophores open the possibility for evaluation of metabolic state at the eye. For interpretation of 2-dimensional measurements of time-resolved auto fluorescence in 2 separate spectral ranges at the human eye, comparing measurements were performed on porcine eyes. Determining excitation and emission spectra, attention was drawn of proof of coenzymes NADH and FAD in isolated anatomical structures cornea, aqueous humor, lens, vitreous, neuronal retina, retinal pigment epithelium (RPE), choroid, and sclera. All these structures exhibit auto fluorescence, highest in lens. Excitation at 350 nm results in local fluorescence maxima at 460 nm, corresponding to NADH, in all structures. This short-wave excitation allows metabolic studies only at the anterior eye, because of the limited transmission of the ocular media. During excitation at 446 nm the existence of FAD is expressed by local fluorescence maxima at 530 nm. The composition fluorescence spectra allow no discrimination between single ocular structures. Approximating the dynamic fluorescence by a double exponential function, the shortest lifetimes were detected in RPE and neuronal retina. The histograms of mean lifetime t M cover each other on lens with cornea and also on sclera with choroid. Despite the lifetimes are close between RPE and neuronal retina, the relative contributions Q I are wide different. The gradient of trend lines in cluster diagrams of amplitudes α II vs. α I allows a discrimination of ocular structures.

Towards microalbuminuria determination on a disposable diagnostic microchip with integrated fluorescence detection based on thin-film organic light emitting diodes.

PubMed

Hofmann, Oliver; Wang, Xuhua; Demello, John C; Bradley, Donal D C; Demello, Andrew J

2005-08-01

As a first step towards a fully disposable stand-alone diagnostic microchip for determination of urinary human serum albumin (HSA), we report the use of a thin-film organic light emitting diode (OLED) as an excitation source for microscale fluorescence detection. The OLED has a peak emission wavelength of 540 nm, is simple to fabricate on flexible or rigid substrates, and operates at drive voltages below 10 V. In a fluorescence assay, HSA is reacted with Albumin Blue 580, generating a strong emission at 620 nm when excited with the OLED. Filter-less discrimination between excitation light and generated fluorescence is achieved through an orthogonal detection geometry. When the assay is performed in 800 microm deep and 800 microm wide microchannels on a poly(dimethylsiloxane)(PDMS) microchip at flow rates of 20 microL min(-1), HSA concentrations down to 10 mg L(-1) can be detected with a linear range from 10 to 100 mg L(-1). This sensitivity is sufficient for the determination of microalbuminuria (MAU), an increased urinary albumin excretion indicative of renal disease (clinical cut-off levels: 15-40 mg L(-1)).

Differences in fluorescence profiles from breast cancer tissues due to changes in relative tryptophan content via energy transfer: tryptophan content correlates with histologic grade and tumor size but not with lymph node metastases

NASA Astrophysics Data System (ADS)

Sordillo, Laura A.; Sordillo, Peter P.; Budansky, Yury; Pu, Yang; Alfano, Robert R.

2014-12-01

The correlation between histologic grade, an increasingly important measure of prognosis for patients with breast cancer, and tryptophan levels from tissues of 15 breast carcinoma patients was investigated. Changes in the relative content of key native organic biomolecule tryptophan were seen from the fluorescence spectra of cancerous and paired normal tissues with excitation wavelengths of 280 and 300 nm. Due to a large spectral overlap and matching excitation-emission spectra, fluorescence resonance energy transfer from tryptophan-donor to reduced nicotinamide adenine dinucleotides-acceptor was noted. We used the ratios of fluorescence intensities at their spectral emission peaks, or spectral fingerprint peaks, at 340, 440, and 460 nm. Higher ratios correlated strongly with high histologic grade, while lower-grade tumors had low ratios. Large tumor size also correlated with high ratios, while the number of lymph node metastases, a major factor in staging, was not correlated with tryptophan levels. High histologic grade correlates strongly with increased content of tryptophan in breast cancer tissues and suggests that measurement of tryptophan content may be useful as a part of the evaluation of these patients.

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

NASA Astrophysics Data System (ADS)

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.

One pot synthesis of intriguing fluorescent carbon dots for sensing and live cell imaging.

PubMed

Jana, Jayasmita; Ganguly, Mainak; Das, Bodhisatwa; Dhara, Santanu; Negishi, Yuichi; Pal, Tarasankar

2016-04-01

We report a simple one-pot synthesis of highly fluorescent carbon dots (CDs) via modified hydrothermal (MHT) treatment of alkaline solution of dopamine and cysteine. These CDs (λex=320 nm, λem=390 nm, and quantum yield ∼ 5.1%) are of ∼ 2-3 nm in diameter. Further attempt of synthesizing CDs in some common water-miscible solvents ends up the fact that the MHT product from acetone medium is nonfluorescent. However, CDs, produced in aqueous medium, are so stable that they can be dried as a deliverable solid (WCD) without any alteration of fluorescing property if reversibly dispersed in water. Fluorescence of WCD is quenched selectively in acetone. Quenching occurs presumably due to the disruption of radiative recombination along with the hindrance in quantum confinement of the emissive energy traps to the particle surface. Successive quenching of fluorescence of WCD in different acetone concentration admixed in water paves the way to selective acetone sensing (LOD=8.75 × 10(-7) M). The synthesized CDs (in aqueous medium) are cytocompatible and are efficient fluorescent probe for cell imaging. Only living cells are recognized exclusively from fluorescence imaging leaving aside dead cells, while cells are treated with CDs. Copyright © 2015 Elsevier B.V. All rights reserved.

Sensitive spectroscopic detection of large and denatured protein aggregates in solution by use of the fluorescent dye Nile red.

PubMed

Sutter, Marc; Oliveira, Sabrina; Sanders, Niek N; Lucas, Bart; van Hoek, Arie; Hink, Mark A; Visser, Antonie J W G; De Smedt, Stefaan C; Hennink, Wim E; Jiskoot, Wim

2007-03-01

The fluorescent dye Nile red was used as a probe for the sensitive detection of large, denatured aggregates of the model protein beta-galactosidase (E. coli) in solution. Aggregates were formed by irreversible heat denaturation of beta-galactosidase below and above the protein's unfolding temperature of 57.4 degrees C, and the presence of aggregates in heated solutions was confirmed by static light scattering. Interaction of Nile red with beta-galactosidase aggregates led to a shift of the emission maximum (lambda (max)) from 660 to 611 nm, and to an increase of fluorescence intensity. Time-resolved fluorescence and fluorescence correlation spectroscopy (FCS) measurements showed that Nile red detected large aggregates with hydrodynamic radii around 130 nm. By steady-state fluorescence measurements, it was possible to detect 1 nM of denatured and aggregated beta-galactosidase in solution. The comparison with size exclusion chromatography (SEC) showed that native beta-galactosidase and small aggregates thereof had no substantial effect on the fluorescence of Nile red. Large aggregates were not detected by SEC, because they were excluded from the column. The results with beta-galactosidase demonstrate the potential of Nile red for developing complementary analytical methods that overcome the size limitations of SEC, and can detect the formation of large protein aggregates at early stages.

UV-Vis spectroscopy and solvatochromism of the tyrosine kinase inhibitor AG-1478.

PubMed

Khattab, Muhammad; Wang, Feng; Clayton, Andrew H A

2016-07-05

The effect of twenty-one solvents on the UV-Vis spectrum of the tyrosine kinase inhibitor AG-1478 was investigated. The absorption spectrum in the range 300-360nm consisted of two partially overlapping bands at approximately 340nm and 330nm. The higher energy absorption band was more sensitive to solvent and exhibited a peak position that varied from 327nm to 336nm, while the lower energy absorption band demonstrated a change in peak position from 340nm to 346nm in non-chlorinated solvents. The fluorescence spectrum of AG-1478 was particularly sensitive to solvent. The wavelength of peak intensity varied from 409nm to 495nm with the corresponding Stokes shift in the range of 64nm to 155nm (4536cm(-1) to 9210cm(-1)). We used a number of methods to assess the relationship between spectroscopic properties and solvent properties. The detailed analysis revealed that for aprotic solvents, the peak position of the emission spectrum in wavenumber scale correlated with the polarity (dielectric constant or ET(30)) of the solvent. In protic solvents, a better correlation was observed between the hydrogen bonding power of the solvent and the position of the emission spectrum. Moreover, the fluorescence quantum yields were larger in aprotic solvents as compared to protic solvents. This analysis underscores the importance of polarity and hydrogen-bonding environment on the spectroscopic properties of AG-1478. These studies will assume relevance in understanding the interaction of AG-1478 in vitro and in vivo. Copyright © 2016 Elsevier B.V. All rights reserved.

A highly selective fluorescent chemosensor for Cu2+ : synthesis and properties of a rhodamine B-containing diarylethene.

PubMed

Xue, Dandan; Zheng, Chunhong; Qu, Shengzu; Liao, Guanming; Fan, Congbin; Liu, Gang; Pu, Shouzhi

2017-06-01

A diarylethene bearing a triazole-linked rhodamine B unit was synthesized. Its fluorescent emission was significantly enhanced in the presence of protons or Cu 2 + due to transformation from the pirocyclic form to open-ring form. The fluorescence was quenched sequentially upon irradiation with 297 nm light based on the intramolecular fluorescence resonance energy transfer mechanism. In an acetonitrile: water binary solvent (1: 1 v/v), the compound showed significant fluorescent enhancement for Cu 2 + compared with a wide range of tested metal ions with a fast response and a limit of detection of 2.86 × 10 -8  mol L -1 . Using Cu 2 + and UV light as the chemical inputs, and fluorescence intensity at 597 nm as the output, a logic gate was developed at the molecular level. Moreover, the compound can be used with a high accuracy to detect Cu 2 + in a natural water sample. Copyright © 2016 John Wiley & Sons, Ltd.

Canopy Level Solar Induced Fluorescence for Vegetation in Controlled Experiments

NASA Technical Reports Server (NTRS)

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 estimates derived using FluorMOD, a radiative transfer model that includes fluorescence properties of foliage, when provided with our supporting measurements. Simulated values for SIF were also compared with similar estimates made over three years for corn (Zea mays L) crops under N treatments (20, 50,70, and 140 kg Nhectare) and with corn and pepper (Capsicum annuum L.) plants provided dimethyl urea (DCMU, 0 and 5 x 10" M) which were grown in pots and grouped in artificial canopies for spectral measurements. For near-surface measurements of healthy and stressed vegetation, red SIF retrieved at 688 nm (023) varied between 2-7 mW/m(exp 2)/nm/sr while far-red SIF retrieved at 760 nm (O2A) varied between 0.5 and 4.0 mW/m(exp 2)/nm/sr. Typical values for the SIF red/far-red ratio ranged between 1.75 and 4.0. Relationships of SIF to spectral reflectance indices and foliar photochemical indices were examined, such as photosynthetic light use efficiency. Initial investigations of the variability in these measurements at the same leaf temperature for sunlit and shaded foliage showed us that shaded foliage produced higher ChlF and light use efficiency. Other factors that affect SIF determinations are discussed. These results will assist in determine the expected intensity of the SIF signal from vegetation near the surface, as well as to identify enhancements needed for FluorMOD (or other such models), and will assist in determining relationships of SIF to reflectance indices and carbon dynamics.

Non-Intrusive Optical Diagnostic Methods for Flowfield Characterization

NASA Technical Reports Server (NTRS)

Tabibi, Bagher M.; Terrell, Charles A.; Spraggins, Darrell; Lee, Ja. H.; Weinstein, Leonard M.

1997-01-01

Non-intrusive optical diagnostic techniques such as Electron Beam Fluorescence (EBF), Laser-Induced Fluorescence (LIF), and Focusing Schlieren (FS) have been setup for high-speed flow characterization and large flowfield visualization, respectively. Fluorescence emission from the First Negative band of N2(+) with the (0,0) vibration transition (at lambda =391.44 nm) was obtained using the EBF technique and a quenching rate of N2(+)* molecules by argon gas was reported. A very high sensitivity FS system was built and applied in the High-Speed Flow Generator (HFG) at NASA LaRC. A LIF system is available at the Advanced Propulsion Laboratory (APL) on campus and a plume exhaust velocity measurement, measuring the Doppler shift from lambda = 728.7 nm of argon gas, is under way.

A novel metronidazole fluorescent nanosensor based on graphene quantum dots embedded silica molecularly imprinted polymer.

PubMed

Mehrzad-Samarin, Mina; Faridbod, Farnoush; Dezfuli, Amin Shiralizadeh; Ganjali, Mohammad Reza

2017-06-15

A novel optical nanosensor for detection of Metronidazole in biological samples was reported. Graphene quantum dots embedded silica molecular imprinted polymer (GQDs-embedded SMIP) was synthesized and used as a selective fluorescent probe for Metronidazole detection. The new synthesized GQDs-embedded SMIP showed strong fluorescent emission at 450nm excited at 365nm which quenched in presence of Metronidazole as a template molecule.. The quenching was proportional to the concentration of Metronidazole in a linear range of at least 0.2μM to 15μM. The limit of detection for metronidazole determination was obtained 0.15μM. The nanosensor successfully worked in plasma matrixes. Copyright © 2016 Elsevier B.V. All rights reserved.

Determination of Ivermectin in Medicated Feeds by Liquid Chromatography with Fluorescence Detection

PubMed Central

2013-01-01

A labour- and time-effective analytical procedure for determination of ivermectin in medicated feed at recommended level of 2.0 mg kg−1 has been developed and validated. The analyte was extracted from grinded feed samples with acetonitrile and derivatisated with N-methylimidazole and trifluoracetic anhydride. The fluorescent derivatives were analysed by liquid chromatography method using C8 column. The isocratic conditions using acetonitrile, methanol, water, and tetrahydrofuran were applied. Fluorescence detection was performed at 365 nm (excitation) and 475 nm (emission) wavelengths. The total analysis time was 10 min. The validation results of the method (within-laboratory reproducibility 4.0% CV, mean recovery 100.1%) confirm the appropriate precision and accuracy of the developed method. PMID:24453835

Light-induced Changes in Allophycocyanin 1

PubMed Central

Ohad, Itzhak; Schneider, Hans-Jörg A. W.; Gendel, Steven; Bogorad, Lawrence

1980-01-01

Several lines of evidence indicate that allophycocyanin is the previously unidentified “phycochrome” observed in extracts of blue-green algae. Fractions containing phycoerythrin, phycocyanin, and allophycocyanin and exhibiting light-induced absorbance changes were prepared from extracts of Nostoc muscorum and Fremyella diplosiphon by isoelectric focusing. Illumination of such fractions with red light (650 nanometers) causes a reduction in absorbance at 620 nm (≃1 to 2%) and an increase at 560 nm. The effect, (previously observed by Björn and Björn [1976 Physiol Plant 36: 297-304]) is reversible, upon illumination with green light (550 nm). Selective immunoprecipitation of the phycobiliproteins indicates that allophycocyanin is the photoresponsive pigment. At pH 4.0 to 4.2, allophycocyanin purified from the same algae or from Phormidium luridum exhibits a light-induced absorbance change at 620 nm, which coincides with its absorption maximum at this pH; the fluorescence emission of allophycocyanin under these conditions is at 647 nm and its S20,w is 2.28, compatible with an α1β1 polypeptide composition. At neutral pH (5.8 to 7.0), allophycocyanin aggregates have a sedimentation coefficient of 4.8 (≃α3β3) and an additional absorption peak at 640 nm appears while that at 620 nm remains unaffected. The fluorescence emission maximum of the larger aggregate is at 667 nm and the light-induced change in its absorption is shifted to 650 nm. The effect of pH changes in the range 4.0 to 7.0 on the spectral and aggregation properties of allophycocyanin is completely reversible. Changes in pH which affect allophycocyanin aggregation have parallel effects on absorption and fluorescence maxima as well as on the light-induced absorbance changes of the biliprotein. No evidence is provided to resolve whether this phycochrome plays the role of an adaptochrome. PMID:16661143

Oral cancer detection based on fluorescence polarization of blood plasma at excitation wavelength 405 nm

NASA Astrophysics Data System (ADS)

Pachaiappan, Rekha; Prakasarao, Aruna; Manoharan, Yuvaraj; Dornadula, Koteeswaran; Singaravelu, Ganesan

2017-02-01

During metabolism the metabolites such as hormones, proteins and enzymes were released in to the blood stream by the cells. These metabolites reflect any change that occurs due to any disturbances in normal metabolic function of the human system. This was well observed with the altered spectral signatures observed with fluorescence spectroscopic technique. Previously many have reported on the significance of native fluorescence spectroscopic method in the diagnosis of cancer. As fluorescence spectroscopy is sensitive and simple, it has complementary techniques such as excitation-emission matrix, synchronous and polarization. The fluorescence polarization measurement provides details about any association or binding reactions and denaturing effects that occurs due to change in the micro environment of cells and tissues. In this study, we have made an attempt in the diagnosis of oral cancer at 405 nm excitation using fluorescence polarization measurement. The fluorescence anisotropic values calculated from polarized fluorescence spectral data of normal and oral cancer subjects yielded a good accuracy when analyzed with linear discriminant analysis based artificial neural network. The results will be discussed in detail.

Three-photon excitation of 2,5-bis(4-biphenyl)oxazole: steady-state and time-resolved intensities and anisotropies

NASA Astrophysics Data System (ADS)

Gryczynski, Ignacy; Malak, Henryk; Hell, Stefan W.; Lakowicz, Joseph R.

1996-10-01

Three-photon excitation of 2,5-bis(4-biphenyl) oxazole (BBO) was observed when it was excited with the fundamental output of a femtosecond Ti:sapphire laser above 820 nm. The emission spectrum of BBO was identical for one-, two-, and three-photon excitation at 320, 640, and 960 nm, respectively. In toluene and triacetin, the emission intensity of BBO depended on the square of the laser power for wavelengths below 820 nm and displayed a sharp transition to a cubic dependence at longer wavelengths. The spatial distribution of the emission of BBO with three- photon excitation was more strongly localized than for two- photon excitation of a coumarin fluorophore at the same wavelength. The same single exponential intensity decay was observed for one-, two-, and three-photon excitation. However, the frequency domain anisotropy decay with three- photon excitation at 960 nm revealed a larger time-zero anisotropy, larger differential polarized phase angle, and larger modulated anisotropy than is possible for two-photon excitation with colinear oscillators. In triacetin, the anisotropy is not constant for three-photon excitation at different wavelengths. Surprisingly, the fluorescence intensities for three-photon excitation were only about 100- fold less than for two-photon excitation. The increasing availability of Ti:sapphire lasers suggests that multiphoton excitation can become a common tool in fluorescence spectroscopy.

Er-doped sesquioxides for 1.5-micron lasers - spectroscopic comparisons

NASA Astrophysics Data System (ADS)

Merkle, Larry D.; Ter-Gabrielyan, Nikolay

2013-05-01

Due to the favorable thermal properties of sesquioxides as hosts for rare earth laser ions, we have recently studied the spectroscopy of Er:Lu2O3 in the 1400-1700 nm wavelength range, and here report its comparison with our earlier results on Er:Y2O3 and Er:Sc2O3. These studies include absorption and fluorescence spectra, fluorescence lifetimes, and inference of absorption and stimulated emission cross sections, all as a function of temperature. At room temperature, optical absorption limits practical laser operation to wavelengths longer than about 1620 nm. In that spectral range, the strongest stimulated emission peak is that at 1665 nm in Er:Sc2O3, with an effective cross section considerably larger than those of Er:Y2O3 and Er:Lu2O3. At 77K, the absorption is weak enough for efficient laser operation at considerably shorter wavelengths, where there are peaks with much larger stimulated emission cross sections. The three hosts all have peaks near 1575-1580 nm with comparably strong cross sections. As we have reported earlier, it is possible to lase even shorter wavelengths efficiently at this temperature, in particular the line at 1558 nm in Er:Sc2O3. Our new spectroscopic studies of Er:Lu2O3 indicate that its corresponding peak, like that of Er:Sc2O3, has a less favorable ratio of stimulated emission to absorption cross sections. Reasons for the differences will be discussed. We conclude that for most operating scenarios, Er:Sc2O3 is the most promising of the Er-doped sesquioxides studied for laser operation around 1.5-1.6 microns.

Photophysical Properties of Organoplatinum(II) Compounds and Derived Self-Assembled Metallacycles and Metallacages: Fluorescence and its Applications.

PubMed

Saha, Manik Lal; Yan, Xuzhou; Stang, Peter J

2016-11-15

Over the past couple of decades, coordination-driven self-assembly has evolved as a broad multidisciplinary domain that not only covers the syntheses of aesthetically pleasing supramolecular architectures but also emerges as a method to form new optical materials, chemical sensors, theranostic agents, and compounds with light-harvesting and emissive properties. The majority of these applications depend upon investigations that reveal the photophysical nature and electronic structure of supramolecular coordination complexes (SCCs), including two-dimensional (2D) metallacycles and three-dimensional (3D) metallacages. As such, well-defined absorption and emission spectra are important for a given SCC to be used for sensing, bioimaging, and other applications with molecular fluorescence being an important component. In this Account, we summarize the photophysical properties of some bis(phosphine)organoplatinum(II) compounds and their discrete SCCs. The platinum(II) based organometallic precursors typically display spectral red-shifts and have low fluorescence quantum yields and short fluorescence lifetimes compared to their organic counterparts because the introduction of metal centers enhances both intersystem crossing (ISC) and intramolecular charge transfer (ICT) processes, which can compete with the fluorescence emissions. Likewise ligands with conjugation can also increase the ICT process; hence the corresponding organoplatinum(II) compounds undergo a further decrease in fluorescence lifetimes. The use of endohedral amine functionalized 120°-bispyridyl ligands can dramatically enhance the emission properties of the resultant organoplatinum(II) based SCCs. As such these SCCs display emissions in the visible region (ca. 400-500 nm) and are significantly red-shifted (ca. 80-100 nm) compared to the ligands. This key feature makes them suitable as supramolecular theranostic agents wherein these unique emission properties provide diagnostic spectroscopic handles and the organoplatinum(II) centers act as potential anticancer agents. Using steady state and time-resolved-spectroscopic techniques and quantum computations in concert, we have determined that the emissive properties stem from the ligand-centered transitions involving π-type molecular orbitals with modest contributions from the metal-based orbitals. The self-assembly and the photophysics of organoplatinum(II) ← 3-substituted pyridyl based SCCs are highly diverse. Subtle changes in the ligands' structures can form molecular congener systems with distinct conformational and photophysical properties. Furthermore, the heterometallic SCCs described herein possess rich photophysical properties and can be used for sensing based applications. Tetraphenylethylene (TPE) based SCCs display emissions in the aggregated state as well as in dilute solutions. This is a unique phenomenon that bridges the aggregation caused quenching (ACQ) and aggregation induced emission (AIE) effects. Moreover, a TPE based metallacage exhibits solvatoluminescence, including white light emission in THF solvent, and can act as a fluorescence-sensor for structurally similar ester compounds.

Micromanipulation and physiological monitoring of cells using two-photon excited fluorescence in cw laser tweezers

NASA Astrophysics Data System (ADS)

Sonek, Gregory J.; Liu, Yagang; Berns, Michael W.; Tromberg, Bruce J.

1996-05-01

We report the observation of two-photon fluorescence excitation and cell confinement, simultaneously, in a continuous-wave (cw) single-beam gradient force optical trap, and demonstrate its use as an in-situ probe to study the physiological state of an optically confined cell sample. At the wavelength of 1064 nm, a single focused gaussian laser beam is used to simultaneously confine, and excite visible fluorescence from, a human sperm cell that has been tagged with propidium iodide, a exogenous fluorescent dye that functions as a viability assay of cellular physiological state. The intensity at the dye peak emission wavelength of 620 nm exhibits a near-square-law dependence on incident trapping beam photon laser power, a behavior consistent with a two-photon absorption process. In addition, for a sperm cell held stationary in the optical tweezers for a period of several minutes at a constant trapping power, red fluorescence emission was observed to increase the time, indicating that the cell has gradually transitioned between a live and dead state. Two-photon excited fluorescence was also observed in chinese hamster ovary cells that were confined by cw laser tweezers and stained with either propidium iodide or Snarf, a pH-sensitive dye probe. These results suggest that, for samples suitably tagged with fluorescent probes and vital stains, optical tweezers can be used to generate their own in-situ diagnostic optical probes of cellular viability or induced photodamage, via two-photon processes.

Use of fluorescence EEM to monitor the removal of emerging contaminants in full scale wastewater treatment plants.

PubMed

Sgroi, Massimiliano; Roccaro, Paolo; Korshin, Gregory V; Greco, Valentina; Sciuto, Sebastiano; Anumol, Tarun; Snyder, Shane A; Vagliasindi, Federico G A

2017-02-05

This study investigated the applicability of different techniques for fluorescence excitation/emission matrices data interpretations, including peak-picking method, fluorescence regional integration and PARAFAC modelling, to act as surrogates in predicting emerging trace organic compounds (ETOrCs) removal during conventional wastewater treatments that usually comprise primary and secondary treatments. Results showed that fluorescence indexes developed using alternative methodologies but indicative of a same dissolved organic matter component resulted in similar predictions of the removal of the target compounds. The peak index defined by the excitation/emission wavelength positions (λ ex/ λ em ) 225/290nm and related to aromatic proteins and tyrosine-like fluorescence was determined to be a particularly suitable surrogate for monitoring ETOrCs that had very high removal rates (average removal >70%) (i.e., triclosan, caffeine and ibuprofen). The peak index defined by λ ex/ λ em =245/440nm and the PARAFAC component with wavelength of the maxima λ ex/ λ em =245, 350/450, both identified as humic-like fluorescence, were found remarkably well correlated with ETOrCs such as atenolol, naproxen and gemfibrozil that were moderately removed (51-70% average removal). Finally, the PARAFAC component with wavelength of the maxima λ ex/ λ em =<240, 315/380 identified as microbial humic-like fluorescence was the only index correlated with the removal of the antibiotic trimethoprim (average removal 68%). Copyright © 2016 Elsevier B.V. All rights reserved.

Synthesis and systematic evaluation of dark resonance energy transfer (DRET)-based library and its application in cell imaging.

PubMed

Su, Dongdong; Teoh, Chai Lean; Kang, Nam-Young; Yu, Xiaotong; Sahu, Srikanta; Chang, Young-Tae

2015-03-01

In this paper, we report a new strategy for constructing a dye library with large Stokes shifts. By coupling a dark donor with BODIPY acceptors of tunable high quantum yield, a novel dark resonance energy transfer (DRET)-based library, named BNM, has been synthesized. Upon excitation of the dark donor (BDN) at 490 nm, the absorbed energy is transferred to the acceptor (BDM) with high efficiency, which was tunable in a broad range from 557 nm to 716 nm, with a high quantum yield of up to 0.8. It is noteworthy to mention that the majority of the non-radiative energy loss of the donor was converted into the acceptor's fluorescence output with a minimum leak of donor emission. Fluorescence imaging tested in live cells showed that the BNM compounds are cell-permeable and can also be employed for live-cell imaging. This is a new library which can be excited through a dark donor allowing for strong fluorescence emission in a wide range of wavelengths. Thus, the BNM library is well suited for high-throughput screening or multiplex experiments in biological applications by using a single laser excitation source. © 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Hyperspectral Imaging and Spectroscopy of Fluorescently Coupled Acyl-CoA: Cholesterol Acyltransferase in Insect Cells

NASA Technical Reports Server (NTRS)

Malak, H.; Mahtani, H.; Herman, P.; Vecer, J.; Lu, X.; Chang, T. Y.; Richmond, Robert C.; Whitaker, Ann F. (Technical Monitor)

2001-01-01

A high-performance hyperspectral imaging module with high throughput of light suitable for low-intensity fluorescence microscopic imaging and subsequent analysis, including single-pixel-defined emission spectroscopy, was tested on Sf21 insect cells expressing green fluorescence associated with recombinant green fluorescent protein linked or not with the membrane protein acyl-CoA:cholesterol acyltransferase. The imager utilized the phenomenon of optical activity as a new technique providing information over a spectral range of 220-1400 nm, and was inserted between the microscope and an 8-bit CCD video-rate camera. The resulting fluorescence image did not introduce observable image aberrations. The images provided parallel acquisition of well resolved concurrent spatial and spectral information such that fluorescence associated with green fluorescent protein alone was demonstrated to be diffuse within the Sf21 insect cell, and that green fluorescence associated with the membrane protein was shown to be specifically concentrated within regions of the cell cytoplasm. Emission spectra analyzed from different regions of the fluorescence image showed blue shift specific for the regions of concentration associated with the membrane protein.

The photochemical trapping rate from red spectral states in PSI-LHCI is determined by thermal activation of energy transfer to bulk chlorophylls.

PubMed

Jennings, Robert C; Zucchelli, Giuseppe; Croce, Roberta; Garlaschi, Flavio M

2003-03-06

The average fluorescence decay lifetimes, due to reaction centre photochemical trapping, were calculated for wavelengths in the 690- to 770-nm interval from the published fluorescence decay-associated emission spectra for Photosystem I (PSI)-light-harvesting complex of Photosystem I (LHCI) [Biochemistry 39 (2000) 6341] at 280 and 170 K. For 280 K, the overall trapping time at 690 nm is 81 ps and increases with wavelength to reach 103 ps at 770 nm. For 170 K, the 690-nm value is 115 ps, increasing to 458 ps at 770 nm. This underlines the presence of kinetically limiting processes in the PSI antenna (diffusion limited). The explanation of these nonconstant values for the overall trapping time band is sought in terms of thermally activated transfer from the red absorbing states to the "bulk" acceptor chlorophyll (chl) states in the framework of the Arrhenius-Eyring theory. It is shown that the wavelength-dependent "activation energies" come out in the range between 1.35 and 2.7 kcal mol(-1), increasing with the emission wavelength within the interval 710-770 nm. These values are in good agreement with the Arrhenius activation energy determined for the steady-state fluorescence yield over the range 130-280 K for PSI-LHCI. We conclude that the variable trapping time in PSI-LHCI can be accounted for entirely by thermally activated transfer from the low-energy chl states to the bulk acceptor states and therefore that the position of the various red states in the PSI antenna seems not to be of significant importance. The analysis shows that the bulk antenna acceptor states are on the low-energy side of the bulk antenna absorption band.

[Determination of cyclamate in complex matrix using HPLC after column derivatization with 4-fluoro-7-nitrobenzofurazan].

PubMed

Hauck, M; Köbler, H

1990-01-01

A method for the analysis of cyclamate in complex foodstuffs has been developed. This method is applicable in strongly coloured and protein-rich foodstuffs. The quantitative determination depends on oxidation of cyclamate to cyclohexylamine and derivatisation with 4-fluoro-7-nitrobenzofuran (NBD-F). The derivatives are analysed by HPLC on a C18: reversed-phase column, their minimal stability being 12 h. There are two possible methods of detection: (a) absorbance at 485 nm and (b) fluorescence with excitation at 485 nm and emission at 530 nm. The detection limit of cyclamate is 5 mg/kg foodstuff, with fluorescence detection 0.4 mg/kg. The recoveries are in the range of 88% to 104%.

Near-infrared luminescence of Bi2ZnOB2O6:Nd3+/PMMA composite

NASA Astrophysics Data System (ADS)

Jaroszewski, K.; Głuchowski, P.; Chrunik, M.; Jastrząb, R.; Majchrowski, A.; Kasprowicz, D.

2018-01-01

Near-infrared luminescence of a novel polymer composite system: PMMA doped with Bi2ZnOB2O6:Nd3+ microparticles, is reported for the first time. Luminescence properties of Bi2ZnOB2O6:Nd3+/PMMA were analyzed on the basis on excitation and emission spectra as well as fluorescence decay profiles. Excitation spectra monitored at 1062 nm (4F3/2 → 4I11/2) indicate numerous bands related to the optical transition of Nd3+ ions: from the 4I9/2 ground state to the 4D3/2, 2P1/2, 2K15/2, 4G7/2 + 4G9/2, 2K13/2, 4G5/2 + 2G7/2, 2H11/2, 4F9/2, 4F7/2 + 4S3/2, 4F5/2 + 2H9/2, 4F3/2 excited states. Many of them may be utilized to excite near-infrared emission of Nd3+ ions. In particular, distinctive Nd3+ emissions of the 4F3/2 → 4I9/2 and 4F3/2 → 4I11/2 transitions were detected, under excitation at 514 nm. The fluorescence decay profiles monitored at 1062 nm, excited at 514 nm, show relatively long emission lifetime of the 4F3/2 → 4I11/2 transition equal to 85 μs. Raman spectroscopy was used to determine vibrational properties and homogeneity of Bi2ZnOB2O6:Nd3+/PMMA composites. The obtained results suggest that Bi2ZnOB2O6:Nd3+/PMMA composite may be applied as an effective source of near-infrared emission in a new integrated optoelectronic devices.

Diphenylacrylonitrile-connected BODIPY dyes: fluorescence enhancement based on dark and AIE resonance energy transfer.

PubMed

Lin, Liangbin; Lin, Xiaoru; Guo, Hongyu; Yang, Fafu

2017-07-19

This study focuses on the construction of novel diphenylacrylonitrile-connected BODIPY dyes with high fluorescence in both solution and an aggregated state by combining DRET and FRET processes in a single donor-acceptor system. The first BODIPY derivatives with one, two, or three AIE-active diphenylacrylonitrile groups were designed and synthesized in moderate yields. Strong fluorescence emissions were observed in the THF solution under excitation at the absorption wavelength of non-emissive diphenylacrylonitrile chromophores, implying the existence of the DRET process between the dark diphenylacrylonitrile donor and the emissive BODIPY acceptor. In the THF/H 2 O solution, the fluorescence intensity of the novel BODIPY derivatives gradually increased under excitation at the absorption wavelength of diphenylacrylonitrile chromophores, suggesting a FRET process between diphenylacrylonitrile and BODIPY moieties. A greater number of diphenylacrylonitrile units led to higher energy-transfer efficiencies. The pseudo-Stokes shift for both DRET and FRET processes was as large as 190 nm.

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

Rout, Dipak; Vijaya, R.; Centre for Lasers and Photonics, Indian Institute of Technology Kanpur, Kanpur 208016

Well-ordered opaline photonic crystals are grown by inward growing self-assembly method from Rhodamine B dye-doped polystyrene colloids. Subsequent to self-assembly, the crystals are infiltrated with gold nanoparticles of 40 nm diameter. Measurements of the stopband features and photoluminescence intensity from these crystals are supplemented by fluorescence decay time analysis. The fluorescence decay times from the dye-doped photonic crystals before and after the infiltration are dramatically different from each other. A lowered fluorescence decay time was observed for the case of gold infiltrated crystal along with an enhanced emission intensity. Double-exponential decay nature of the fluorescence from the dye-doped crystal gets convertedmore » into single-exponential decay upon the infiltration of gold nanoparticles due to the resonant radiative process resulting from the overlap of the surface plasmon resonance with the emission spectrum. The influence of localized surface plasmon due to gold nanoparticles on the increase in emission intensity and decrease in decay time of the emitters is established.« less

[Fluorescence excitation-emission matrix spectroscopy of CDOM from Yundang Lagoon and its indication for organic pollution].

PubMed

Zhuo, Jian-Fu; Guo, Wei-Dong; Deng, Xun; Zhang, Zhi-Ying; Xu, Jing; Huang, Ling-Feng

2010-06-01

Fluorescence excitation-emission matrix spectroscopy (EEMs) combined with absorption spectroscopy were applied to study the optical properties of CDOM samples from highly-polluted Yundang Lagoon in Xiamen in order to demonstrate the feasibility of using these spectral properties as a tracer of the degree of organic pollution in similar polluted coastal waters. Surface water samples were collected from 13 stations 4 times during April and May, 2008. Parallel factor analysis (PARAFAC) model was used to resolve the EEMs of CDOM. Five separate fluorescent components were identified, including two humic-like components (C1: 240, 325/422 nm; C5: 260, 380/474 nm), two protein-like components (C2: 225, 275/350 nm; C4: 240, 300/354 nm) and one xenobiotic-like component (C3: 225/342 nm), which could be used as a good tracer for the input of the anthropogenic organic, pollutants. The concentrations of component C3 and dissolved organic carbon (DOC) are much higher near the inlet of sewage discharge, demonstrating that the discharge of surrounding sewage is a major source of organic pollutants in Yundang Lagoon. CDOM absorption coefficient alpha (280) and the score of humic-like component C1 showed significant linear relationships with COD(Mn), and a strong positive correlation was also found between the score of protein-like component C2 and BOD5. This suggested that the optical properties of CDOM may provide a fast in-situ way to monitor the variation of the water quality in Yundang Lagoon and that of similar polluted coastal waters.

A turn-on fluorescence chemosensor based on a tripodal amine [tris(pyrrolyl-α-methyl)amine]-rhodamine conjugate for the selective detection of zinc ions.

PubMed

Balamurugan, Rathinam; Chang, Wen-I; Zhang, Yandison; Fitriyani, Sri; Liu, Jui-Hsiang

2016-09-21

A novel tetradendate ligand derived from a tris(pyrrolyl-α-methyl)amine (H3tpa) and rhodamine-based conjugate (PR) has been designed for use as a sensor, synthesized and characterized spectroscopically. PR {(tris(5-rhodamineiminopyrrol-2-ylmethyl)amine)} serves as a selective colorimetric as well as a fluorescent chemosensor for Zn(2+) in acetonitrile/water (1 : 1, v/v). In the presence of Zn(2+), PR exhibited obvious absorption (558 nm) and emission (577 nm) peaks whose intensity increased along with increasing Zn(2+) concentrations. Titration experiments revealed that a large excess of Zn(2+) was required to saturate the absorption (λmax) and emission intensities. Upon the addition of 1000 equivalents of Zn(2+), the fluorescence intensity of the PR underwent an ∼500-fold increase (Φf = 0.34) with the emission maximum at 580 nm. These kinetics studies demonstrated that the absorption and emission changes were proportional to the Zn(2+) concentration. The color of the solution changed from colorless to a dark pink color. The fluorescence of the PR-Zn(2+) complex can be reversibly restored by using ammonium water or by heating. Competitive ion tests revealed that the intensity of PR-Zn(2+) was not suppressed by excess amounts of other metal ions. The counter anions did not exert obvious influences on the absorption and emission profiles. (1)H-NMR and FT-IR spectroscopic investigations of PR and PR-Zn(2+) revealed that the pyrrole motifs, -C[double bond, length as m-dash]N- groups and spirolactam of rhodamine B are capable of coordinating cation guest species. Because each arm of the tripodal ligand tautomerizes independently, only moderate fluorescence enhancement could be seen until all three -C[double bond, length as m-dash]N- groups were coordinated by zinc, which may be due to the spirolactam ring opening mechanism of the rhodamine unit. Once all three -C[double bond, length as m-dash]N- groups were locked by coordinating with excess of Zn(2+), the isomerization was arrested, and PR exhibited highly enhanced fluorescence. In addition, energy optimized structures of PR were found to be cage-like by Gaussian 09, further supporting that it can access a large excess of Zn(2+). Intriguingly, imaging of HeLa cells by using a confocal microscope revealed that this PR probe could be used for biological applications.

The binding of terbium ions to tubulin induces ring formation.

PubMed

Monasterio, O; Acoria, M; Díaz, M A; Lagos, R

1993-02-01

The intrinsic fluorescence excitation and emission spectra of chicken brain tubulin showed the characteristic tryptophan fluorescence. The emission spectrum of Tb3+ in the presence of tubulin and GTP excited at 295 nm, showed four peaks, with the maxima at 490, 545, and 586 nm and a minor peak around 620 nm. Titration of tubulin with Tb3+ was followed by the increment in luminescence at 545 nm and showed a sigmoidal curve where the initial lag interval and the maximal luminescence intensity depended on tubulin concentration. The presence of Mg2+, Co2+, and Zn2+ diminished both the sigmoidicity of the curve and the maximal luminescence intensity. Titration of tubulin with Tb3+ also produced a sigmoidal increase in turbidity, which was shifted to the left with respect to the luminescence curve. The dependence of turbidity on the wavelength of the Tb(3+)-induced polymers revealed that the large structures formed were not microtubules. Electron microscopy of the aggregates induced by Tb3+ showed mainly a lattice of double rings with side-by-side contacts. These results indicate that Tb3+ induces principally double ring formation and that these rings (33 +/- 2 nm external diameter) aggregate in large-ordered arrays. The luminescence of Tb3+ seems to be induced mainly by the aggregation of rings.

Laser Ablated Carbon Nanodots for Light Emission.

PubMed

Reyes, Delfino; Camacho, Marco; Camacho, Miguel; Mayorga, Miguel; Weathers, Duncan; Salamo, Greg; Wang, Zhiming; Neogi, Arup

2016-12-01

The synthesis of fluorescent carbon dots-like nanostructures (CNDs) obtained through the laser ablation of a carbon solid target in liquid environment is reported. The ablation process was induced in acetone with laser pulses of 1064, 532, and 355 nm under different irradiation times. Close-spherical amorphous CNDs with sizes between 5 and 20 nm, whose abundance strongly depends on the ablation parameters were investigated using electron microscopy and was confirmed using absorption and emission spectroscopies. The π- π* electronic transition at 3.76 eV dominates the absorption for all the CNDs species synthesized under different irradiation conditions. The light emission is most efficient due to excitation at 3.54 eV with the photoluminescence intensity centered at 3.23 eV. The light emission from the CNDs is most efficient due to ablation at 355 nm. The emission wavelength of the CNDs can be tuned from the near-UV to the green wavelength region by controlling the ablation time and modifying the ablation and excitation laser wavelength.

Selective detection of Co2+ by fluorescent nano probe: Diagnostic approach for analysis of environmental samples and biological activities

NASA Astrophysics Data System (ADS)

Mahajan, Prasad G.; Dige, Nilam C.; Desai, Netaji K.; Patil, Shivajirao R.; Kondalkar, Vijay V.; Hong, Seong-Karp; Lee, Ki Hwan

2018-06-01

Nowadays scientist over the world are engaging to put forth improved methods to detect metal ion in an aqueous medium based on fluorescence studies. A simple, selective and sensitive method was proposed for detection of Co2+ ion using fluorescent organic nanoparticles. We synthesized a fluorescent small molecule viz. 4,4‧-{benzene-1,4-diylbis-[(Z)methylylidenenitrilo]}dibenzoic acid (BMBA) to explore its suitability as sensor for Co2+ ion and biocompatibility in form of nanoparticles. Fluorescence nanoparticles (BMBANPs) prepared by simple reprecipitation method. Aggregation induced enhanced emission of BMBANPs exhibits the narrower particle size of 68 nm and sphere shape morphology. The selective fluorescence quenching was observed by addition of Co2+ and does not affected by presence of other coexisting ion solutions. The photo-physical properties, viz. UV-absorption, fluorescence emission, and lifetime measurements are in support of ligand-metal interaction followed by static fluorescence quenching phenomenon in emission of BMBANPs. Finally, we develop a simple analytical method for selective and sensitive determination of Co2+ ion in environmental samples. The cell culture E. coli, Bacillus sps., and M. tuberculosis H37RV strain in the vicinity of BMBANPs indicates virtuous anti-bacterial and anti-tuberculosis activity which is of additional novel application shown by prepared nanoparticles.

Spatial-Temporal Mapping of the T Cell Receptor NF-kappaB Signaling Pathway

DTIC Science & Technology

2006-05-30

Lasos), a 700/488-nm excitation filter and a 500– 550-nm emission filter. Multiphoton PA-GFP activation was performed using the Aim software bleach ...Bcl10 was fused to a PA-GFP [65], and MALT1 was fused to the reef coral fluorescent protein, monomeric Kusabira-Orange (mKO) [70]. D10 T-cells

Mn-doped Zinc Sulphide nanocrystals for immunofluorescent labeling of epidermal growth factor receptors on cells and clinical tumor tissues

NASA Astrophysics Data System (ADS)

J, Aswathy; V, Seethalekshmy N.; R, Hiran K.; R, Bindhu M.; K, Manzoor; Nair, Shantikumar V.; Menon, Deepthy

2014-11-01

The field of molecular detection and targeted imaging has evolved considerably with the introduction of fluorescent semiconductor nanocrystals. Manganese-doped zinc sulphide nanocrystals (ZnS:Mn NCs), which are widely used in electroluminescent displays, have been explored for the first time for direct immunofluorescent (IF) labeling of clinical tumor tissues. ZnS:Mn NCs developed through a facile wet chemistry route were capped using amino acid cysteine, conjugated to streptavidin and thereafter coupled to biotinylated epidermal growth factor receptor (EGFR) antibody utilizing the streptavidin-biotin linkage. The overall conjugation yielded stable EGFR antibody conjugated ZnS:Mn NCs (EGFR ZnS:Mn NCs) with a hydrodynamic diameter of 65 ± 15 nm, and having an intense orange-red fluorescence emission at 598 nm. Specific labeling of EGF receptors on EGFR+ve A431 cells in a co-culture with EGFR-ve NIH3T3 cells was demonstrated using these nanoprobes. The primary antibody conjugated fluorescent NCs could also clearly delineate EGFR over-expressing cells on clinical tumor tissues processed by formalin fixation as well as cryopreservation with a specificity of 86% and accuracy of 88%, in comparison to immunohistochemistry. Tumor tissues labeled with EGFR ZnS:Mn NCs showed good fluorescence emission when imaged after storage even at 15 months. Thus, ZnS nanobioconjugates with dopant-dependent and stable fluorescence emission show promise as an efficient, target-specific fluorophore that would enable long term IF labeling of any antigen of interest on clinical tissues.

Detection of lead in brass by laser-induced breakdown spectroscopy combined with laser-induced fluorescence

NASA Astrophysics Data System (ADS)

Goueguel, Christian; Laville, Stéphane; Loudyi, Hakim; Chaker, Mohamed; Sabsabi, Mohamad; Vidal, François

2008-06-01

Laser-Induced Breakdown Spectroscopy (LIBS) technique combined with Laser-Induced Fluorescence (LIF) is known to be a high sensitivity and high selectivity analytical technique. Although sub-ppm limits of detection (LoD) have already been demonstrated, there is still a constant and urgent need to reach lower LoDs. Here, we report results obtained for the detection of lead trace in brass samples. The plasma was produced by a Q-switched Nd:YAG laser at 1064 nm and then re-excited by a nanosecond optical parametric oscillator (OPO) laser tuned at 283.31 nm. Emission from Pb atoms was then observed at 405.78 nm. The experiments were performed in air at atmospheric pressure. We found out that the optimal conditions were obtained for an ablation fluence of 2-3 J/cm2 and inter-pulse delay of 8-10 μs. Also, excitation energy of about 200 μJ was required to maximize the Pb(I) 405.78 nm emission. Using the LIBS-LIFS technique, the LoD was estimated to be about 180 ppb over 100 laser shots, which corresponds to an improvement of about two orders of magnitude with that obtained using conventional LIBS.

Highly sensitive and selective cartap nanosensor based on luminescence resonance energy transfer between NaYF4:Yb,Ho nanocrystals and gold nanoparticles.

PubMed

Wang, Zhijiang; Wu, Lina; Shen, Baozhong; Jiang, Zhaohua

2013-09-30

Fluorescent detection is an attractive method for the detection of toxic chemicals. However, most chemosensors that are currently utilized in fluorescent detection are based on organic dyes or quantum dots, which suffer from instability, high background noise and interference from organic impurities in solution, which can also be excited by UV radiation. In the present research, we developed a novel NaYF4:Yb,Ho/Au nanocomposite-based chemosensor with high sensitivity (10 ppb) and selectivity over competing analytes for the detection of the insecticide cartap. This nanosensor is excited with a 970-nm laser instead of UV radiation to give an emission peak at 541 nm. In the presence of cartap, the nanocomposites aggregate, resulting in enhanced luminescence resonance energy transfer between the NaYF4:Yb,Ho nanocrystals and the gold nanoparticles, which decreases the emission intensity at 541 nm. The relative luminescence intensity at 541 nm has a linear relationship with the concentration of cartap in the solution. Based on this behavior, the developed nanosensor successfully detected cartap in farm produce and water samples with satisfactory results. Copyright © 2013 Elsevier B.V. All rights reserved.

Developing LED UV fluorescence sensors for online monitoring DOM and predicting DBPs formation potential during water treatment.

PubMed

Li, Wen-Tao; Jin, Jing; Li, Qiang; Wu, Chen-Fei; Lu, Hai; Zhou, Qing; Li, Ai-Min

2016-04-15

Online monitoring dissolved organic matter (DOM) is urgent for water treatment management. In this study, high performance size exclusion chromatography with multi-UV absorbance and multi-emission fluorescence scans were applied to spectrally characterize samples from 16 drinking water sources across Yangzi River and Huai River Watersheds. The UV absorbance indices at 254 nm and 280 nm referred to the same DOM components and concentration, and the 280 nm UV light could excite both protein-like and humic-like fluorescence. Hence a novel UV fluorescence sensor was developed out using only one UV280 light-emitting diode (LED) as light source. For all samples, enhanced coagulation was mainly effective for large molecular weight biopolymers; while anion exchange further substantially removed humic substances. During chlorination tests, UVA280 and UVA254 showed similar correlations with yields of disinfection byproducts (DBPs); the humic-like fluorescence obtained from LED sensors correlated well with both trihalomethanes and haloacetic acids yields, while the correlation between protein-like fluorescence and trihalomethanes was relatively poor. Anion exchange exhibited more reduction of DBPs yields as well as UV absorbance and fluorescence signals than enhanced coagulation. The results suggest that the LED UV fluorescence sensors are very promising for online monitoring DOM and predicting DBPs formation potential during water treatment. Copyright © 2016 Elsevier Ltd. All rights reserved.

Fluorescence properties of human teeth and dental calculus for clinical applications

NASA Astrophysics Data System (ADS)

Lee, Yong-Keun

2015-04-01

Fluorescent emission of human teeth and dental calculus is important for the esthetic rehabilitation of teeth, diagnosis of dental caries, and detection of dental calculus. The purposes of this review were to summarize the fluorescence and phosphorescence of human teeth by ambient ultraviolet (UV) light, to investigate the clinically relevant fluorescence measurement methods in dentistry, and to review the fluorescence of teeth and dental calculus by specific wavelength light. Dentine was three times more phosphorescent than enamel. When exposed to light sources containing UV components, the fluorescence of human teeth gives them the quality of vitality, and fluorescent emission with a peak of 440 nm is observed. Esthetic restorative materials should have fluorescence properties similar to those of natural teeth. Based on the fluorescence of teeth and restorative materials as determined with a spectrophotometer, a fluorescence parameter was defined. As to the fluorescence spectra by a specific wavelength, varied wavelengths were investigated for clinical applications, and several methods for the diagnosis of dental caries and the detection of dental calculus were developed. Since fluorescent properties of dental hard tissues have been used and would be expanded in diverse fields of clinical practice, these properties should be investigated further, embracing newly developed optical techniques.

Fluorescence properties of human teeth and dental calculus for clinical applications.

PubMed

Lee, Yong-Keun

2015-04-01

Fluorescent emission of human teeth and dental calculus is important for the esthetic rehabilitation of teeth, diagnosis of dental caries, and detection of dental calculus. The purposes of this review were to summarize the fluorescence and phosphorescence of human teeth by ambient ultraviolet (UV) light, to investigate the clinically relevant fluorescence measurement methods in dentistry, and to review the fluorescence of teeth and dental calculus by specific wavelength light. Dentine was three times more phosphorescent than enamel. When exposed to light sources containing UV components, the fluorescence of human teeth gives them the quality of vitality, and fluorescent emission with a peak of 440 nm is observed. Esthetic restorative materials should have fluorescence properties similar to those of natural teeth. Based on the fluorescence of teeth and restorative materials as determined with a spectrophotometer, a fluorescence parameter was defined. As to the fluorescence spectra by a specific wavelength, varied wavelengths were investigated for clinical applications, and several methods for the diagnosis of dental caries and the detection of dental calculus were developed. Since fluorescent properties of dental hard tissues have been used and would be expanded in diverse fields of clinical practice, these properties should be investigated further, embracing newly developed optical techniques.

Protein labeling with red squarylium dyes for analysis by capillary electrophoresis with laser-induced fluorescence detection.

PubMed

Yan, Weiying; Sloat, Amy L; Yagi, Shigeyuki; Nakazumi, Hiroyuki; Colyer, Christa L

2006-04-01

Two new red luminescent asymmetric squarylium dyes (designated "Red-1c and Red-3") have been shown to exhibit absorbance shifts to longer wavelengths upon the addition of protein, along with a concomitant increase in fluorescence emission. Specifically, the absorbance maxima for Red-1c and Red-3 dyes are 607 and 622 nm, respectively, in the absence of HSA, and 642 and 640 nm in the presence of HSA, making the excitation of their protein complexes feasible with inexpensive and robust diode lasers. Fluorescence emission maxima, in the presence of HSA, are 656 and 644 nm for Red-1c and Red-3, respectively. Because of the inherently low fluorescence of the dyes in their free state, Red-1c and Red-3 were used as on-column labels (that is, with the dye incorporated into the separation buffer), thus eliminating the need for sample derivatization prior to injection and separation. A comparison of precolumn and on-column labeling of proteins with these squarylium dyes revealed higher efficiencies and greater sensitivities for on-column labeling, which, when conducted with a basic, high-salt content buffer, permitted baseline resolution of a mixture of five model proteins. LOD for model proteins, such as transferrin, alpha-lactalbumin, BSA, and beta-lactoglobulin A and B, labeled with these dyes and analyzed by CE with LIF detection (CE-LIF) were found to be dependent upon dye concentration and solution pH, and are as low as 5 nM for BSA. Satisfactory linear relationships between peak height (or peak area) and protein concentration were obtained by CE-LIF for this on-column labeling method with Red-3 and Red-1c.

Investigating the 3.3 micron infrared fluorescence from naphthalene following ultraviolet excitation

NASA Technical Reports Server (NTRS)

Williams, Richard M.; Leone, Stephen R.

1994-01-01

Polycyclic aromatic hydrocarbon (PAH) type molecules are proposed as the carriers of the unidentified infrared (UIR) bands. Detailed studies of the 3.3 micrometer infrared emission features from naphthalene, the simplest PAH, following ultraviolet laser excitation are used in the interpretation of the 3.29 micrometer (3040 cm(sup -1)) UIR band. A time-resolved Fourier transform spectrometer is used to record the infrared emission spectrum of gas-phase naphthalene subsequent to ultraviolet excitation facilitated by an excimer laser operated at either 193 nm or 248 nm. The emission spectra differ significantly from the absorption spectrum in the same spectral region. Following 193 nm excitation the maximum in the emission profile is red-shifted 45 cm(sup -1) relative to the absorption maximum; a 25 cm(sup -1) red-shift is observed after 248 nm excitation. The red-shifting of the emission spectrum is reduced as collisional and radiative relaxation removes energy from the highly vibrationally excited molecules. Coupling between the various vibrational modes is thought to account for the differences between absorption and emission spectra. Strong visible emission is also observed following ultraviolet excitation. Visible emission may play an important role in the rate of radiative relaxation, which according to the interstellar PAH hypothesis occurs only by the slow emission of infrared photons. Studying the visible emission properties of PAH type molecules may be useful in the interpretation of the DIB's observed in absorption.

Discrimination of geographical origin and detection of adulteration of kudzu root by fluorescence spectroscopy coupled with multi-way pattern recognition

NASA Astrophysics Data System (ADS)

Hu, Leqian; Ma, Shuai; Yin, Chunling

2018-03-01

In this work, fluorescence spectroscopy combined with multi-way pattern recognition techniques were developed for determining the geographical origin of kudzu root and detection and quantification of adulterants in kudzu root. Excitation-emission (EEM) spectra were obtained for 150 pure kudzu root samples of different geographical origins and 150 fake kudzu roots with different adulteration proportions by recording emission from 330 to 570 nm with excitation in the range of 320-480 nm, respectively. Multi-way principal components analysis (M-PCA) and multilinear partial least squares discriminant analysis (N-PLS-DA) methods were used to decompose the excitation-emission matrices datasets. 150 pure kudzu root samples could be differentiated exactly from each other according to their geographical origins by M-PCA and N-PLS-DA models. For the adulteration kudzu root samples, N-PLS-DA got better and more reliable classification result comparing with the M-PCA model. The results obtained in this study indicated that EEM spectroscopy coupling with multi-way pattern recognition could be used as an easy, rapid and novel tool to distinguish the geographical origin of kudzu root and detect adulterated kudzu root. Besides, this method was also suitable for determining the geographic origin and detection the adulteration of the other foodstuffs which can produce fluorescence.

A Quninolylthiazole Derivatives as an ICT-Based Fluorescent Probe of Hg(II) and its Application in Ratiometric Imaging in Live HeLa Cells.

PubMed

Bai, Jian-Ying; Xie, Yu-Zhong; Wang, Chang-Jiang; Fang, Shu-Qing; Cao, Lin-Nan; Wang, Ling-Li; Jin, Jing-Yi

2018-05-28

As a structural analogue of pyridylthiazole, 2-(2-benzothiazoyl)-phenylethynylquinoline (QBT) was designed as a fluorescent probe for Hg(II) based on an intramolecular charge transfer (ICT) mechanism. The compound was synthesized in three steps starting from 6-bromo-2-methylquinoline, with moderate yield. Corresponding studies on the optical properties of QBT indicate that changes in the fluorescence ratio of QBT in response to Hg(II) could be quantified based on dual-emission changes. More specifically, the emission spectrum of QBT before and after interactions with Hg(II) exhibited a remarkable red shift of about 120 nm, which is rarely reported in ICT-based fluorescent sensors. Finally, QBT was applied in the two-channel imaging of Hg(II) in live HeLa cells.

Emission behaviors of unsymmetrical 1,3-diaryl-β-diketones: A model perfectly disclosing the effect of molecular conformation on luminescence of organic solids

NASA Astrophysics Data System (ADS)

Cheng, Xiao; Li, Feng; Han, Shenghua; Zhang, Yufei; Jiao, Chuanjun; Wei, Jinbei; Ye, Kaiqi; Wang, Yue; Zhang, Hongyu

2015-03-01

A series of unsymmetrical 1,3-diaryl-β-diketones 1-6 displaying molecular conformation-dependent fluorescence quantum yields have been synthesized. Crystals with planar molecular conformation such as 1, 2, 3 and 4 are highly fluorescent (φf: 39-53%), and the one holding slightly twisted conformation (5) is moderately luminescent (φf = 17%), while crystal 6 possessing heavily bent structure is completely nonluminous (φf ~ 0). The distinct fluorescence efficiencies are ascribed to their different molecular conformations, since all the crystals hold the same crystal system, space group and crystal packing structures. Additionally, the fluorescent crystals 1-5 display low threshold amplified spontaneous emission (ASE) with small full widths at half-maximum (FWHM: 3-7 nm), indicating their potential as candidates for organic crystal lasing devices.

A flexible 1,8-naphthyridyl derivative and its Zn(II) complexes: synthesis, structures, spectroscopic properties and recognition of Cd(II).

PubMed

Zhang, Hui-Miao; Fu, Wen-Fu; Gan, Xin; Xu, Yan-Qing; Wang, Jun; Xu, Quan-Qing; Chi, Shao-Ming

2008-12-21

A flexible ligand bis(7-methyl-1,8-naphthyridine-2-ylamino)methane (), having kappa(4)-chelating and kappa(2)-bridging modes, and its intriguing structural complexes of Zn(II) with mu-OH, kappa(1)-OAc, mu-kappa(1)-OAc and mu-kappa(2)-OAc ligands, [Zn(2)()(2)(OH)](ClO(4))(3) (), [Zn(4)()(2)(OAc)(6)(OH)(2)].CH(2)Cl(2) (.CH(2)Cl(2)) and [Zn(5)()(2)(OAc)(10)](n).4nH(2)O (.4H(2)O) were synthesized and their structures were determined by X-ray crystallography. These compounds exhibited intense blue fluorescent emissions with a lambda(max) in the range of 380-410 nm in CH(2)Cl(2), CH(3)CN and CH(3)OH solutions, and solid-state emissions centered at 416, 463, 490 and 451 nm were observed for the compounds , , and at room temperature, respectively. The investigated fluorescence properties of associated with various metal ions showed that the fluorescence enhancement of with Cd(II) was more sensitive than with other interfering cations.

Selective identification and quantification of saccharin by liquid chromatography and fluorescence detection.

PubMed

Bruno, Sergio N F; Cardoso, Carlos R; Maciel, Márcia Mosca A; Vokac, Lidmila; da Silva Junior, Ademário I

2014-09-15

High-pressure liquid chromatography with ultra-violet detection (HPLC-UV) is one of the most commonly used methods to identify and quantify saccharin in non-alcoholic beverages. However, due to the wide variety of interfering UV spectra in saccharin-containing beverage matrices, the same method cannot be used to measure this analyte accurately. We have developed a new, highly effective method to identify and quantify saccharin using HPLC with fluorescence detection (HPLC-FLD). The excitation wavelength (250 nm) and emission wavelength (440 nm) chosen increased selectivity for all matrices and ensured few changes were required in the mobile phase or other parameters. The presence of saccharin in non-diet beverages - a fraud commonly used to replace more expensive sucrose - was confirmed by comparing coincident peaks as well as the emission spectra of standards and samples. Copyright © 2014 Elsevier Ltd. All rights reserved.

Time-resolved laser-induced fluorescence system

NASA Astrophysics Data System (ADS)

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

2006-02-01

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

Fluorescent Nanodiamonds in Biomedical Applications.

PubMed

Mitura, Katarzyna Anna; Włodarczyk, Elżbieta

2018-04-18

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

Detection of large deletion in human BRCA1 gene in human breast carcinoma MCF-7 cells by using DNA-Silver Nanoclusters

NASA Astrophysics Data System (ADS)

Borghei, Yasaman-Sadat; Hosseini, Morteza; Ganjali, Mohammad Reza

2018-01-01

Here we describe a label-free detection strategy for large deletion mutation in breast cancer (BC) related gene BRCA1 based on a DNA-silver nanocluster (NC) fluorescence upon recognition-induced hybridization. The specific hybridization of DNA templated silver NCs fluorescent probe to target DNAs can act as effective templates for enhancement of AgNCs fluorescence, which can be used to distinguish the deletion of BRCA1 due to different fluorescence intensities. Under the optimal conditions, the fluorescence intensity of the DNA-AgNCs at emission peaks around 440 nm (upon excitation at 350 nm) increased with the increasing deletion type within a dynamic range from 1.0 × 10-10 to 2.4 × 10-6 M with a detection limit (LOD) of 6.4 × 10-11 M. In this sensing system, the normal type shows no significant fluorescence; on the other hand, the deletion type emits higher fluorescence than normal type. Using this nanobiosensor, we successfully determined mutation using the non-amplified genomic DNAs that were isolated from the BC cell line.

A Cu²⁺-selective fluorescent chemosensor based on BODIPY with two pyridine ligands and logic gate.

PubMed

Huang, Liuqian; Zhang, Jing; Yu, Xiaoxiu; Ma, Yifan; Huang, Tianjiao; Shen, Xi; Qiu, Huayu; He, Xingxing; Yin, Shouchun

2015-06-15

A novel near-infrared fluorescent chemosensor based on BODIPY (Py-1) has been synthesized and characterized. Py-1 displays high selectivity and sensitivity for sensing Cu(2+) over other metal ions in acetonitrile. Upon addition of Cu(2+) ions, the maximum absorption band of Py-1 in CH3CN displays a red shift from 603 to 608 nm, which results in a visual color change from pink to blue. When Py-1 is excited at 600 nm in the presence of Cu(2+), the fluorescent emission intensity of Py-1 at 617 nm is quenched over 86%. Notably, the complex of Py-1-Cu(2+) can be restored with the introduction of EDTA or S(2-). Consequently, an IMPLICATION logic gate at molecular level operating in fluorescence mode with Cu(2+) and S(2-) as chemical inputs can be constructed. Finally, based on the reversible and reproducible system, a nanoscale sequential memory unit displaying "Writing-Reading-Erasing-Reading" functions can be integrated. Copyright © 2015 Elsevier B.V. All rights reserved.

Novel fluorescence adjustable photonic crystal materials

NASA Astrophysics Data System (ADS)

Zhu, Cheng; Liu, Xiaoxia; Ni, Yaru; Fang, Jiaojiao; Fang, Liang; Lu, Chunhua; Xu, Zhongzi

2017-11-01

Novel photonic crystal materials (PCMs) with adjustable fluorescence were fabricated by distributing organic fluorescent powders of Yb0.2Er0.4Tm0.4(TTA)3Phen into the opal structures of self-assembled silica photonic crystals (PCs). Via removing the silica solution in a constant speed, PCs with controllable thicknesses and different periodic sizes were obtained on glass slides. Yb0.2Er0.4Tm0.4(TTA)3Phen powders were subsequently distributed into the opal structures. The structures and optical properties of the prepared PCMs were investigated. Finite-difference-time-domain (FDTD) calculation was used to further analyze the electric field distributions in PCs with different periodic sizes while the relation between periodic sizes and fluorescent spectra of PCMs was discussed. The results showed that the emission color of the PCMs under irradiation of 980 nm laser can be easily adjusted from green to blue by increasing the periodic size from 250 to 450 nm.

Fluorescence properties of 6-aryl-2‧-deoxy-furanouridine and -pyrrolocytidine and their derivatives

NASA Astrophysics Data System (ADS)

Ro, Jong Jin; Go, Gui Han; Wilhelmsson, L. Marcus; Hyean Kim, Byeang

2018-01-01

2‧-deoxyfuranouridine derivatives presenting various aryl groups have been synthesized through Cu(I)-catalyzed intramolecular cyclizations. Moreover, corresponding pyrrolo-dC derivatives have been synthesized and both families of compounds thoroughly characterized using UV/vis and fluorescence spectroscopy as well as time-dependent density functional theory calculations. The photophysical characterization, show that our newly synthesized derivatives of the important pyrrolo-dC family have high fluorescence quantum yields (QYs) and brightness values. Pyrrolo-dC derivative, 3a, shows an environment sensitive QY of up to >60% and brightness of almost 3000, in low polarity solvents and excitation and emission maxima between 365-381 nm and 479-510 nm, respectively, in solvents of different polarities. Two other derivatives, 3b and 3c, show high QYs and brightness values of up to 3300 that are fairly insensitive to their microenvironment. These promising photophysical features suggest future applicability as fluorescent nucleobase analogs.

Distance-dependent plasmon-enhanced fluorescence of upconversion nanoparticles using polyelectrolyte multilayers as tunable spacers.

PubMed

Feng, Ai Ling; You, Min Li; Tian, Limei; Singamaneni, Srikanth; Liu, Ming; Duan, Zhenfeng; Lu, Tian Jian; Xu, Feng; Lin, Min

2015-01-14

Lanthanide-doped upconversion nanoparticles (UCNPs) have attracted widespread interests in bioapplications due to their unique optical properties by converting near infrared excitation to visible emission. However, relatively low quantum yield prompts a need for developing methods for fluorescence enhancement. Plasmon nanostructures are known to efficiently enhance fluorescence of the surrounding fluorophores by acting as nanoantennae to focus electric field into nano-volume. Here, we reported a novel plasmon-enhanced fluorescence system in which the distance between UCNPs and nanoantennae (gold nanorods, AuNRs) was precisely tuned by using layer-by-layer assembled polyelectrolyte multilayers as spacers. By modulating the aspect ratio of AuNRs, localized surface plasmon resonance (LSPR) wavelength at 980 nm was obtained, matching the native excitation of UCNPs resulting in maximum enhancement of 22.6-fold with 8 nm spacer thickness. These findings provide a unique platform for exploring hybrid nanostructures composed of UCNPs and plasmonic nanostructures in bioimaging applications.

Study of absorption and re-emission processes in a ternary liquid scintillation system

NASA Astrophysics Data System (ADS)

Xiao, Hua-Lin; Li, Xiao-Bo; Zheng, Dong; Cao, Jun; Wen, Liang-Jian; Wang, Nai-Yan

2010-11-01

Liquid scintillators are widely used as the neutrino target in neutrino experiments. The absorption and emission of different components of a ternary liquid scintillator (Linear Alkyl Benzene (LAB) as the solvent, 2,5-diphenyloxazole (PPO) as the fluor and p-bis-(o-methylstyryl)-benzene (bis-MSB) as wavelength shifter) are studied. It is shown that the absorption of this liquid scintillator is dominant by LAB and PPO at wavelengths less than 349 nm, and the absorption by bis-MSB becomes prevalent at the wavelength larger than 349 nm. The fluorescence quantum yields, which are the key parameters to model the absorption and re-emission processes in large liquid scintillation detectors, are measured.

Ratiometric Fluorescent Detection of Pb2+ by FRET-Based Phthalocyanine-Porphyrin Dyads.

PubMed

Zhang, Dongli; Zhu, Mengliang; Zhao, Luyang; Zhang, Jinghui; Wang, Kang; Qi, Dongdong; Zhou, Yang; Bian, Yongzhong; Jiang, Jianzhuang

2017-12-04

Sensitive and selective detection of Pb 2+ is a very worthwhile endeavor in terms of both human health and environmental protection, as the heavy metal is fairly ubiquitous and highly toxic. In this study, we designed phthalocyanine-porphyrin (Pc-Por) heterodyads, namely, H 2 Pc-α-ZnPor (1) and H 2 Pc-β-ZnPor (2), by connecting a zinc(II) porphyrin moiety to the nonperipheral (α) or peripheral (β) position of a metal-free phthalocyanine moiety. Upon excitation at the porphyrin Soret region (420 nm), both of the dyads exhibited not only a porphyrin emission (605 nm) but also a phthalocyanine emission (ca. 700 nm), indicating the occurrence of intramolecular fluorescence resonance energy transfer (FRET) processes from the porphyrin donor to the phthalocyanine acceptor. The dyads can selectively bind Pb 2+ in the phthalocyanine core leading to a red shift of the phthalocyanine absorption and thus a decrease of spectral overlap between the porphyrin emission and phthalocyanine absorption, which in turn suppresses the intramolecular FRET. In addition, the binding of Pb 2+ can highly quench the emission of phthalocyanine by heavy-metal ion effects. The synergistic coupled functions endow the dyads with remarkable ratiometric fluorescent responses at two distinct wavelengths (F 605 /F 703 for 1 and F 605 /F 700 for 2). The emission intensity ratio increased as a linear function to the concentration of Pb 2+ in the range of 0-4.0 μM, whereas the detection limits were determined to be 3.4 × 10 -9 and 2.2 × 10 -8 M for 1 and 2, respectively. Furthermore, by comparative study of 1 and 2, the effects of distance and relative orientation between Pc and ZnPor fluorophores on the FRET efficiency and sensing performance were highlighted, which is helpful for further optimizing such FRET systems.

Autofluorescence of human cells in vitro as a biomarker of their metabolic activity

NASA Astrophysics Data System (ADS)

Dobrzyńska, Monika; Stepińska, Małgorzata; Lewandowski, Rafał; Gietka, Andrzej; Łapiński, Mariusz P.; Trafny, ElŻbieta A.

2016-12-01

Autofluorescence (AF) is the natural emission of light by intrinsic fluorophores. Oxidized mitochondrial flavins, lipofuscin and reduced nicotinamideadenine dinucleotide phosphate (NAD(P)H) are the main sources of the autofluorescence in cells upon excitation with visible light. The aim of the study was to investigate changes in the metabolism of four cell lines by monitoring their autofluorescence with a microplate reader. Autofluorescence intensities of cells were collected at two wavelengths for the excitation and fluorescence emission: for endogenous NAD(P)H at 366/450 nm, for the oxidized flavoproteins and lipofuscin at 460/540 nm. Human mesenchymal stem cells (hMSC), epithelial cells from mammary gland (MCF 10A), breast ductal carcinoma (T-47D) prostate carcinoma (DU-145) were observed daily for 16 days. The level of NAD(P)H autofluorescence did not differ among the cell lines investigated. The significant increase in oxidized flavoproteins fluorescence intensity was recorded for hMSC and ranged from 140 to 175% of control. During 28 days differentiation process, the NAD(P)H, FAD and lipofuscin fluorescence intensities were recorded every day, and the redox ratio was then calculated. The redox ratio gradually decreased during the last eight days of osteogenesis and adipogenesis. Therefore, in our opinion the NAD(P)H, FAD, and lipofuscin fluorescence emission at the wavelengths selected are the optimal parameters to be collected during the differentiation process in order to monitor the metabolism of hMSC undergoing structural and morphological changes.

Photoluminescence Enhancement of Silole-Capped Silicon Quantum Dots Based on Förster Resonance Energy Transfer.

PubMed

Kim, Seongwoong; Kim, Sungsoo; Ko, Young Chun; Sohn, Honglae

2015-07-01

Photoluminescent porous silicon were prepared by an electrochemical etch of n-type silicon under the illumination with a 300 W tungsten filament bulb for the duration of etch. The red photoluminescence emitting at 650 nm with an excitation wavelength of 450 nm is due to the quantum confinement of silicon quantum dots in porous silicon. HO-terminated red luminescent PS was obtained by an electrochemical treatment of fresh PS with the current of 150 mA for 60 seconds in water and sodium chloride. As-prepared PS was sonicated, fractured, and centrifuged in toluene solution to obtain photoluminescence silicon quantum dots. Dichlorotetraphenylsilole exhibiting an emission band at 520 nm was reacted with HO-terminated silicon quantum dots to give a silole-capped silicon quantum dots. The optical characterization of silole-derivatized silicon quantum dots was investigated by UV-vis and fluorescence spectrometer. The fluorescence emission efficiency of silole-capped silicon quantum dots was increased by about 2.5 times due to F6rster resonance energy transfer from silole moiety to silicon quantum dots.

[Resolving excitation emission matrix spectroscopy of estuarine CDOM with parallel factor analysis and its application in organic pollution monitoring].

PubMed

Guo, Wei-Dong; Huang, Jian-Ping; Hong, Hua-Sheng; Xu, Jing; Deng, Xun

2010-06-01

The distribution and estuarine behavior of fluorescent components of chromophoric dissolved organic matter (CDOM) from Jiulong Estuary were determined by fluorescence excitation emission matrix spectroscopy (EEMs) combined with parallel factor analysis (PARAFAC). The feasibility of these components as tracers for organic pollution in estuarine environments was also evaluated. Four separate fluorescent components were identified by PARAFAC, including three humic-like components (C1: 240, 310/382 nm; C2: 230, 250, 340/422 nm; C4: 260, 390/482 nm) and one protein-like components (C3: 225, 275/342 nm). These results indicated that UV humic-like peak A area designated by traditional "peak-picking method" was not a single peak but actually a combination of several fluorescent components, and it also had inherent links to so-called marine humic-like peak M or terrestrial humic-like peak C. Component C2 which include peak M decreased with increase of salinity in Jiulong Estuary, demonstrating that peak M can not be thought as the specific indicator of the "marine" humic-like component. Two humic-like components C1 and C2 showed additional behavior in the turbidity maximum region (salinity < 6) and then conservative mixing behavior for the rest estuarine region, while humic-like components C4 showed conservative mixing behavior for the whole estuarine region. However, the protein-like component C3 showed nonconservative mixing behavior, suggesting it had autochthonous estuarine origin. EEMs-PARAFAC can provide fluorescent fingerprint to differentiate the DOM features for three tributaries of Jiulong River. The observed linear relationships between humic-like components and absorption coefficient a (280) with chemical oxygen demand (COD) and biological oxygen demand (BOD5) suggest that the optical properties of CDOM may provide a fast in-situ way to monitor the variation of the degree of organic pollution in estuarine environments.

Fluorescent cooling of objects exposed to sunlight – The ruby example

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

Berdahl, Paul; Chen, Sharon S.; Destaillats, Hugo

Various pigments are used to formulate desirable non-white colors that stay cooler in the sun than alternatives, which is particularly useful for hot climate areas. These cool pigments provide a high near-infrared (NIR) reflectance in the solar infrared range of 700-2500 nm, and also a color specified by a reflectance spectrum in the 400-700 nm visible range. Still cooler materials can be formulated by also utilizing the phenomenon of fluorescence (photoluminescence). Ruby, Al 2O 3 :Cr, is a prime example, with efficient emission in the deep red (~694 nm) and near infrared (700-800 nm). A layer of synthetic ruby crystalsmore » on a white surface having an attractive red color can remain cooler in the sun than conventional red materials. Ruby particles can also be used as a red/pink pigment. Increasing the Cr:Al ratio produces a stronger (darker) pigment but doping above ~3 wt% Cr 2O 3 causes concentration quenching of the fluorescence. The system quantum efficiency for lightly doped ruby-pigmented coatings over white is high, 0.83 ± 0.10.« less

A two-photon laser induced fluorescence diagnostic with improved sensitivity, localization, and measurement rate

NASA Astrophysics Data System (ADS)

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.

Enhancement of Single Molecule Fluorescence Signals by Colloidal Silver Nanoparticles in Studies of Protein Translation

PubMed Central

Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S.; Goldman, Yale E.

2011-01-01

Metal enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold respectively. Fluorescence intensity fluctuations above shot noise, at 0.1 – 5 Hz, were greater on silver particles. Overall signal to noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G. PMID:21158483

Enhancement of single-molecule fluorescence signals by colloidal silver nanoparticles in studies of protein translation.

PubMed

Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S; Goldman, Yale E

2011-01-25

Metal-enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold, respectively. Fluorescence intensity fluctuations above shot noise, at 0.1-5 Hz, were greater on silver particles. Overall signal-to-noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G.

PARAFAC Modeling of Irradiation- and Oxidation-Induced Changes in Fluorescent Dissolved Organic Matter Extracted from Poultry Litter.

PubMed

Mangalgiri, Kiranmayi P; Timko, Stephen A; Gonsior, Michael; Blaney, Lee

2017-07-18

Parallel factor analysis (PARAFAC) applied to fluorescence excitation emission matrices (EEMs) allows quantitative assessment of the composition of fluorescent dissolved organic matter (DOM). In this study, we fit a four-component EEM-PARAFAC model to characterize DOM extracted from poultry litter. The data set included fluorescence EEMs from 291 untreated, irradiated (253.7 nm, 310-410 nm), and oxidized (UV-H 2 O 2 , ozone) poultry litter extracts. The four components were identified as microbial humic-, terrestrial humic-, tyrosine-, and tryptophan-like fluorescent signatures. The Tucker's congruence coefficients for components from the global (i.e., aggregated sample set) model and local (i.e., single poultry litter source) models were greater than 0.99, suggesting that the global EEM-PARAFAC model may be suitable to study poultry litter DOM from individual sources. In general, the transformation trends of the four fluorescence components were comparable for all poultry litter sources tested. For irradiation at 253.7 nm, ozonation, and UV-H 2 O 2 advanced oxidation, transformation of the humic-like components was slower than that of the tryptophan-like component. The opposite trend was observed for irradiation at 310-410 nm, due to differences in UV absorbance properties of components. Compared to the other EEM-PARAFAC components, the tyrosine-like component was fairly recalcitrant in irradiation and oxidation processes. This novel application of EEM-PARAFAC modeling provides insight into the composition and fate of agricultural DOM in natural and engineered systems.

Bifunctional Diaminoterephthalate Fluorescent Dye as Probe for Cross-Linking Proteins.

PubMed

Wallisch, Melanie; Sulmann, Stefan; Koch, Karl-Wilhelm; Christoffers, Jens

2017-05-11

Diaminoterephthalates are fluorescent dyes and define scaffolds, which can be orthogonally functionalized at their two carboxylate residues with functional residues bearing task specific reactive groups. The synthesis of monofunctionalized dyes with thiol groups for surface binding, an azide for click chemistry, and a biotinoylated congener for streptavidin binding is reported. Two bifunctionalized dyes were prepared: One with an azide for click chemistry and a biotin for streptavidin binding, the other with a maleimide for reaction with thiol and a cyclooctyne moiety for ligation with copper-free click chemistry. In general, the compounds are red to orange, fluorescent materials with an absorption at about 450 nm and an emission at 560 nm with quantum yields between 2-41 %. Of particular interest is the maleimide-functionalized compound, which shows low fluorescence quantum yield (2 %) by itself. After addition of a thiol, the fluorescence is "turned on"; quantum yield 41 %. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

Spectroscopy characterization and quantum yield determination of quantum dots

NASA Astrophysics Data System (ADS)

Contreras Ortiz, S. N.; Mejía Ospino, E.; Cabanzo, R.

2016-02-01

In this paper we show the characterization of two kinds of quantum dots: hydrophilic and hydrophobic, with core and core/shell respectively, using spectroscopy techniques such as UV-Vis, fluorescence and Raman. We determined the quantum yield in the quantum dots using the quinine sulphate as standard. This salt is commonly used because of its quantum yield (56%) and stability. For the CdTe excitation, we used a wavelength of 549nm and for the CdSe/ZnS excitation a wavelength of 527nm. The results show that CdSe/ZnS (49%) has better fluorescence, better quantum dots, and confirm the fluorescence result. The quantum dots have shown a good fluorescence performance, so this property will be used to replace dyes, with the advantage that quantum dots are less toxic than some dyes like the rhodamine. In addition, in this work we show different techniques to find the quantum dots emission: fluorescence spectrum, synchronous spectrum and Raman spectrum.

Imaging intracellular pH in live cells with a genetically encoded red fluorescent protein sensor.

PubMed

Tantama, Mathew; Hung, Yin Pun; Yellen, Gary

2011-07-06

Intracellular pH affects protein structure and function, and proton gradients underlie the function of organelles such as lysosomes and mitochondria. We engineered a genetically encoded pH sensor by mutagenesis of the red fluorescent protein mKeima, providing a new tool to image intracellular pH in live cells. This sensor, named pHRed, is the first ratiometric, single-protein red fluorescent sensor of pH. Fluorescence emission of pHRed peaks at 610 nm while exhibiting dual excitation peaks at 440 and 585 nm that can be used for ratiometric imaging. The intensity ratio responds with an apparent pK(a) of 6.6 and a >10-fold dynamic range. Furthermore, pHRed has a pH-responsive fluorescence lifetime that changes by ~0.4 ns over physiological pH values and can be monitored with single-wavelength two-photon excitation. After characterizing the sensor, we tested pHRed's ability to monitor intracellular pH by imaging energy-dependent changes in cytosolic and mitochondrial pH.

A coumarin-derived Cu2 +-fluorescent chemosensor and its direct application in aqueous media

NASA Astrophysics Data System (ADS)

Mergu, Naveen; Kim, Myeongjin; Son, Young-A.

2018-01-01

A novel coumarin-based receptor bearing a benzohydrazide (FCBH) was developed as a fluorescent chemosensor with high selectivity toward Cu2 +. The sensor was successfully applied to the monitoring of Cu2 + in aqueous solution. After the addition of Cu2 + to FCBH, the color of the solution changed from greenish-yellow to red, and the absorption band at 457 nm red-shifted to 517 nm. The fluorescent green color of FCBH disappeared and the fluorescence emission was completely quenched in the presence of Cu2 +. Upon the addition of Cu2 +, deprotonation of FCBH occurred, and a 1:1 metal-ligand complex formed. DFT theoretical investigation was carried out to understand the behavior of the sensing probe toward Cu2 +. Additionally, the quenched fluorescence of the FCBH-Cu2 + complex was restored upon the addition of CN- ions. The possible sensing mechanism of FCBH toward Cu2 + was derived from experimental and theoretical examinations.

A fluorescent probe for the efficient discrimination of Cys, Hcy and GSH based on different cascade reactions.

PubMed

Li, Ying; Liu, Weimin; Zhang, Panpan; Zhang, Hongyan; Wu, Jiasheng; Ge, Jiechao; Wang, Pengfei

2017-04-15

A fluorescent probe (1) for distinguishing amongst biothiols, including cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), is developed based on different cascade reactions. The key design feature of fluorescent probe 1 is the integration of two potential reaction groups for the thiol and amino groups of biothiols in one molecule. By reacting with the halogen atom and α, β-unsaturated malonitrile in probe 1, Cys, Hcy and GSH can generate a total of three main products with distinct photophysical properties. Probe 1 shows a strong fluorescence turn-on response to Cys with blue-green emission by using an excitation wavelength of 390nm. At an excitation wavelength of 500nm, probe 1 responds to GSH over Cys and Hcy and emits strong orange fluorescence. The discrimination of biothiols can be demonstrated by cell imaging experiments, indicating that probe 1 can be a useful tool for the selective imaging of Cys and GSH in living cells. Copyright © 2016 Elsevier B.V. All rights reserved.

Effect of solvents on the fluorescence spectra of bacterial luciferase

NASA Astrophysics Data System (ADS)

Sukovataya, Irina E.; Tyulkova, Natalya A.; Kaykova, Elisaveta V.

2006-08-01

Bacteria luciferases catalyze the oxidation reaction of the long-chain aliphatic aldehyde and reduced flavinmononucleotide involving molecular oxygen to a respective fatty acid emitting light quanta in the visible spectrum. Fluorescence emission of luciferases from Photobacterium leiognathi dissolved in organic solvent-water mixtures was investigated. Methanol, acetone, dimethyl sulfoxide and formamide were used as organic solvents. As the methanol and acetone concentration is increased the emission maximum peak is decrease. In contrast, with dimethyl sulfoxide and formamide addition induced a increasing of the emission maximum intensity. The values of wavelength maximum (λ max) at the addition of this solvent can shows the spectra shifted to the red by about 12 nm. These increasing in the fluorescence intensity and in the λ max may be due to luciferase denaturation, resulting from the more intensive contact of chromospheres of luciferase with the solvent. At all used concentrations of methanol, acetone and formamide the shape of the fluorescence spectra was not changed. These studies demonstrate that the luciferase tryptophan fluorescence is sensitive to changes of physical-chemical property of enzyme environment. A comparison of activation/inactivation and fluorescence spectra of luciferase in methanol or acetone solutions shows that the extent of inactivation is larger than the extent of fluorescence changes at the same methanol or acetone concentration.

A highly selective colorimetric and ratiometric fluorescent chemodosimeter for detection of fluoride ions based on 1,8-naphthalimide derivatives.

PubMed

Kai, Yumei; Hu, Yonghong; Wang, Kai; Zhi, Wenbiao; Liang, Mengmeng; Yang, Wenge

2014-01-24

A high selective colorimetric and ratiometric fluorescent probe based on 4-hydroxy-1, 8-naphthalimide was designed and synthesized to detect fluoride ions (F(-)). The sensing behavior of this probe was studied by UV-visible and fluorescence spectroscopy. The probe displays an 110 nm red-shift of fluorescence emission and the color changes from colorless to yellow by virtue of the strong affinity of F(-) toward silicon which can act as a new visual sensor for F(-). Copyright © 2013 Elsevier B.V. All rights reserved.

Laser-induced fluorescence spectroscopy of the secondary cataract

NASA Astrophysics Data System (ADS)

Maslov, N. A.; Larionov, P. M.; Rozhin, I. A.; Druzhinin, I. B.; Chernykh, V. V.

2016-06-01

Excitation-emission matrices of laser-induced fluorescence of lens capsule epithelium, the lens nucleus, and the lens capsule are investigated. A solid-state laser in combination with an optical parametric generator tunable in the range from 210 to 350 nm was used for excitation of fluorescence. The spectra of fluorescence of all three types of tissues exhibit typical features that are specific to them and drastically differ from one another. This effect can be used for intrasurgical control of presence of residual lens capsule epithelium cells in the capsular bag after surgical treatment of a cataract.

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

Lin, C.T.

Linear and nonlinear photochemistries of 1,4-diazabicyclo(2.2.2)octane (DABCO) are investigated at room temperature by using ArF (193 nm) and KrF (248 nm) lasers. With an unfocused beam geometry, DABCO vapor displays a strong fluorescence when excited at 248 nm, but it shows no detectable emission with 193-nm excitation. The linear photochemistry quantum yield for DABCO is determined as phi/sub p/(248nm) approx. 0.1 and phi/sub p/(193 nm) approx. 0.3. The main stable photochemical products are analyzed as C/sub 2/H/sub 4/ and C/sub 2/H/sub 2/ for 248- and 193-nm excitation, respectively. When focused beam excitation is used, both ArF and KrF lasers dissociatemore » DABCO molecules and give three strong radical emissions of CN*(B vector /sup 2/..sigma.. ..-->.. X vector /sup 2/ ..sigma../sup +/), CH*(A vector /sup 2/..delta.. ..-->.. X vector /sup 2/II), and C/sub 2/*(D vector /sup 3/II/sub g/ ..-->.. a vector /sup 3/II/sub u/). The time behavior, the laser power dependence, and the sample pressure dependence of these emissive radicals are examined. The possible mechanisms for the Rydberg state photochemistry of DABCO are discussed.« less

Micelle-induced versatile performance of amphiphilic intramolecular charge-transfer fluorescent molecular sensors.

PubMed

Wang, Jiaobing; Qian, Xuhong; Qian, Junhong; Xu, Yufang

2007-01-01

A series of amphiphilic intramolecular charge-transfer fluorescent molecular sensors AS1-3, equipped with a rod-shaped hydrophobic 2-phenylbenzoxazole fluorophore and a hydrophilic tetraamide Hg(2+)-ion receptor, have been prepared. These sensor molecules could be incorporated into the hydrophobic sodium dodecyl sulfate (SDS) micelle, which is confirmed by the clear spectral blue shift and emission enhancement observed at the critical micelle concentration of SDS. Systematic examination of the sensor-Hg(2+) complexation, by using both UV/visible and fluorescence spectroscopy, indicates that SDS significantly modulates both the binding event and signal transformation of these sensor molecules. The potential advantages are fourfold: 1) SDS substantially increases the Hg(2+)-ion association constant and results in an amplified sensitivity. 2) SDS initiates spectral features which facilitate Hg(2+)-ion analysis, for example, in addition to the strengthened fluorescence of the free sensors AS1-3, the original "on-off" response of AS2 toward the Hg(2+) ion is transformed into a self-calibrated two-wavelength ratiometric signal, while for AS3, Hg(2+)-ion complexation in the presence of SDS results in a 180 nm blue shift, which is preferred to the 51 nm spectral shift obtained without SDS. 3) Thermoreversible tuning of the dynamic detection range is realized. 4) Highly specific Hg(2+)-ion identification could be achieved by using the SDS-induced fingerprint emission (358 nm) of the AS2-Hg(2+) complex. Altogether, this work demonstrates a convenient and powerful strategy that remarkably elevates the performance of a given fluorescent molecular sensor. It also implies that for a specific utilization, much attention should be paid to the microenvironment in which the sensor resides, as the behavior of the sensor might be different from that in the bulk solution.

Norcyanine dyes with benzo[c,d]indolium moiety: Spectral sensitivity with pH change for fluorescence pH imaging in living cells.

PubMed

Guan, Li; Liu, Qi; Zhang, Borui; Wang, Lanying

2017-01-01

Fluorescence pH imaging in living cells is a rapidly expanding research direction, however, it relies on the development of pH-sensitive fluorescent imaging agents. Here four norcyanine dyes with benzo[c,d]indolium moiety, exhibiting high spectral sensitivity with pH changes, were synthesized for fluorescence pH imaging in living cells, and characterized by 1 H NMR, 13 C NMR, IR, UV-Vis and HRMS. The investigation of their spectral properties in methanol and water showed that the absorption and emission maxima were in the region 488-618nm and 583-651nm, respectively, and four dyes exhibited high photostability. The pH spectral titrations showed that selective dye D1 had pH-dependent absorption spectral changes within the pH range of 2.4 to 9.4, and high fluorescent spectral sensitivity at pH5.0-8.0, with a pK a of 5.0. A cell association study indicated that dye D1 exhibited no or mild cytotoxicity at the application dose and duration, and could be accumulated in cells and mainly distributed in the cytoplasm, giving red fluorescence imaging. In particular, dye D1 could achieve pH-dependent fluorescence imaging in living cells with the increase of pH from 3.0 to 8.0, at excitation wavelength of 543nm and receiving wavelength of 655-755nm, which was valuable for studying the weak acidic, neutral and weak alkaline biological tissue compartments. Copyright © 2016 Elsevier B.V. All rights reserved.

Pharmacokinetics of endogenous porphyrins induced by 5-aminolevulinic acid as observed by means of laser-induced fluorescence from several organs of tumor-bearing mice

NASA Astrophysics Data System (ADS)

Sroka, Ronald; Baumgartner, Reinhold; Beyer, Wolfgang; Gossner, Liebwin; Sassy, T.; Stocker, Susanne

1995-04-01

Photodynamic therapy (PDT) and photodynamic diagnosis (PDD) add support to efficient treatment modalities of superficial and early stage cancer. Recently 5-aminolevulinic acid (5-ALA), a precursor of hemoglobin in the hem biosynthetic pathway, was used to stimulate endogenous porphyrin production. The time dependency of 5-ALA induced porphyrin fluorescence has been investigated on several normal tissues as well as on a tumor in an in-vivo tumor model (human gastrointestinal adenocarcinoma Grade II, UICC IIa). 5-ALA has been administered intravenously at a concentration of 50 mg/(kg bw). With respect to a certain time schedule the animals were sacrificed and 12 different organs as well as the tumor were removed. Using laser-induced fluorescence techniques the emission spectra in the range of (lambda) equals (550-750) nm were detected from the tissues after excitation with light of the wavelength (lambda) equals (411 +/- 4) nm. For quantitative evaluation the integral fluorescence intensity at (lambda) equals (635 +/- 2) nm of the porphyrin specific spectra has been determined. All tissues showed porphyrin fluorescence, while brightest fluorescence has been detected from the tumor. With respect to the other tissues the relative tumor selectivity showed a maximum ratio at 406 h post injection. The kinetics of the porphyrin fluorescence intensity of the organs follow different time dependencies. Simple mathematical pharmacokinetic models are developed and discussed.

Optical Band Gap Alteration of Graphene Oxide via Ozone Treatment.

PubMed

Hasan, Md Tanvir; Senger, Brian J; Ryan, Conor; Culp, Marais; Gonzalez-Rodriguez, Roberto; Coffer, Jeffery L; Naumov, Anton V

2017-07-25

Graphene oxide (GO) is a graphene derivative that emits fluorescence, which makes GO an attractive material for optoelectronics and biotechnology. In this work, we utilize ozone treatment to controllably tune the band gap of GO, which can significantly enhance its applications. Ozone treatment in aqueous GO suspensions yields the addition/rearrangement of oxygen-containing functional groups suggested by the increase in vibrational transitions of C-O and C=O moieties. Concomitantly it leads to an initial increase in GO fluorescence intensity and significant (100 nm) blue shifts in emission maxima. Based on the model of GO fluorescence originating from sp 2 graphitic islands confined by oxygenated addends, we propose that ozone-induced functionalization decreases the size of graphitic islands affecting the GO band gap and emission energies. TEM analyses of GO flakes confirm the size decrease of ordered sp 2 domains with ozone treatment, whereas semi-empirical PM3 calculations on model addend-confined graphitic clusters predict the inverse dependence of the band gap energies on sp 2 cluster size. This model explains ozone-induced increase in emission energies yielding fluorescence blue shifts and helps develop an understanding of the origins of GO fluorescence emission. Furthermore, ozone treatment provides a versatile approach to controllably alter GO band gap for optoelectronics and bio-sensing applications.

Concentration and size dependence of peak wavelength shift on quantum dots in colloidal suspension

NASA Astrophysics Data System (ADS)

Rinehart, Benjamin S.; Cao, Caroline G. L.

2016-08-01

Quantum dots (QDs) are semiconductor nanocrystals that have significant advantages over organic fluorophores, including their extremely narrow Gaussian emission bands and broad absorption bands. Thus, QDs have a wide range of potential applications, such as in quantum computing, photovoltaic cells, biological sensing, and electronics. For these applications, aliasing provides a detrimental effect on signal identification efficiency. This can be avoided through characterization of the QD fluorescence signals. Characterization of the emissivity of CdTe QDs as a function of concentration (1 to 10 mg/ml aqueous) was conducted on 12 commercially available CdTe QDs (emission peaks 550 to 730 nm). The samples were excited by a 50-mW 405-nm laser with emission collected via a free-space CCD spectrometer. All QDs showed a redshift effect as concentration increased. On average, the CdTe QDs exhibited a maximum shift of +35.6 nm at 10 mg/ml and a minimum shift of +27.24 nm at 1 mg/ml, indicating a concentration dependence for shift magnitude. The concentration-dependent redshift function can be used to predict emission response as QD concentration is changed in a complex system.

Bright optical centre in diamond with narrow, highly polarised and nearly phonon-free fluorescence at room temperature

NASA Astrophysics Data System (ADS)

John, Roger; Lehnert, Jan; Mensing, Michael; Spemann, Daniel; Pezzagna, Sébastien; Meijer, Jan

2017-05-01

Using shallow implantation of ions and molecules with masses centred at 27 atomic mass units (amu) in diamond, a new artificial optical centre with unique properties has been created. The centre shows a linearly polarised fluorescence with a main narrow emission line mostly found at 582 nm, together with a weak vibronic sideband at room temperature. The fluorescence lifetime is ∼2 ns and the brightest centres are more than three times brighter than the nitrogen-vacancy centres. A majority of the centres shows stable fluorescence whereas some others present a blinking behaviour, at faster or slower rates. Furthermore, a second kind of optical centre has been simultaneously created in the same diamond sample, within the same ion implantation run. This centre has a narrow zero-phonon line (ZPL) at ∼546 nm and a broad phonon sideband at room temperature. Interestingly, optically detected magnetic resonance (ODMR) has been measured on several single 546 nm centres and two resonance peaks are found at 0.99 and 1.27 GHz. In view of their very similar ODMR and optical spectra, the 546 nm centre is likely to coincide with the ST1 centre, reported once (with a ZPL at 550 nm), but of still unknown nature. These new kinds of centres are promising for quantum information processing, sub-diffraction optical imaging or use as single-photon sources.

Detection of microbial biofilms on food processing surfaces: hyperspectral fluorescence imaging study

NASA Astrophysics Data System (ADS)

Jun, Won; Kim, Moon S.; Chao, Kaunglin; Lefcourt, Alan M.; Roberts, Michael S.; McNaughton, James L.

2009-05-01

We used a portable hyperspectral fluorescence imaging system to evaluate biofilm formations on four types of food processing surface materials including stainless steel, polypropylene used for cutting boards, and household counter top materials such as formica and granite. The objective of this investigation was to determine a minimal number of spectral bands suitable to differentiate microbial biofilm formation from the four background materials typically used during food processing. Ultimately, the resultant spectral information will be used in development of handheld portable imaging devices that can be used as visual aid tools for sanitation and safety inspection (microbial contamination) of the food processing surfaces. Pathogenic E. coli O157:H7 and Salmonella cells were grown in low strength M9 minimal medium on various surfaces at 22 +/- 2 °C for 2 days for biofilm formation. Biofilm autofluorescence under UV excitation (320 to 400 nm) obtained by hyperspectral fluorescence imaging system showed broad emissions in the blue-green regions of the spectrum with emission maxima at approximately 480 nm for both E. coli O157:H7 and Salmonella biofilms. Fluorescence images at 480 nm revealed that for background materials with near-uniform fluorescence responses such as stainless steel and formica cutting board, regardless of the background intensity, biofilm formation can be distinguished. This suggested that a broad spectral band in the blue-green regions can be used for handheld imaging devices for sanitation inspection of stainless, cutting board, and formica surfaces. The non-uniform fluorescence responses of granite make distinctions between biofilm and background difficult. To further investigate potential detection of the biofilm formations on granite surfaces with multispectral approaches, principal component analysis (PCA) was performed using the hyperspectral fluorescence image data. The resultant PCA score images revealed distinct contrast between biofilms and granite surfaces. This investigation demonstrated that biofilm formations on food processing surfaces, even for background materials with heterogeneous fluorescence responses, can be detected. Furthermore, a multispectral approach in developing handheld inspection devices may be needed to inspect surface materials that exhibit non-uniform fluorescence.

Two-step excitation and blue fluorescence under continuous-wave pumping in Nd:YLF

NASA Technical Reports Server (NTRS)

Fan, T. Y.; Byer, Robert L.

1986-01-01

Near-UV and blue fluorescence from the 4D3/2 and 4D5/2 manifolds in Nd:YLF has been observed at room temperature under CW pumping by a rhodamine 590 dye laser. Excitation to these manifolds is attributed to two-step excitation involving excited-state absorption from the 4F3/2 metastable level. A similar phenomenon has also been observed in Nd:YAG and Nd:glass. The effective excited-state absorption cross section is measured to be (2 + or - 1) x 10 to the -20th sq cm at 587.4 nm in the pi polarization, and the peak effective stimulated emission cross section is measured to be 5 x 10 to the -20th sq cm at 411.7 nm, also in the pi polarization. Estimated laser threshold at 411.7 nm for two-step pumping at 587.4 nm is 70 mW.

Spectral analysis of tissues from patients with cancer using a portable spectroscopic diagnostic ratiometer unit

NASA Astrophysics Data System (ADS)

Sordillo, Laura A.; Pu, Yang; Sordillo, Peter P.; Budansky, Yury; Alfano, R. R.

2014-05-01

Spectral profiles of tissues from patients with breast carcinoma, malignant carcinoid and non-small cell lung carcinoma were acquired using native fluorescence spectroscopy. A novel spectroscopic ratiometer device (S3-LED) with selective excitation wavelengths at 280 nm and 335 nm was used to produce the emission spectra of the key biomolecules, tryptophan and NADH, in the tissue samples. In each of the samples, analysis of emission intensity peaks from biomolecules showed increased 340 nm/440 nm and 340 nm/460 nm ratios in the malignant samples compared to their paired normal samples. This most likely represented increased tryptophan to NADH ratios in the malignant tissue samples compared to their paired normal samples. Among the non-small cell lung carcinoma and breast carcinomas, it appeared that tumors of very large size or poor differentiation had an even greater increase in the 340 nm/440 nm and 340 nm/460 nm ratios. In the samples of malignant carcinoid, which is known to be a highly metabolically active tumor, a marked increase in these ratios was also seen.

High-Performance Blue-Excitable Yellow Phosphor Obtained from an Activated Solvochromic Bismuth-Fluorophore Metal–Organic Framework

DOE PAGES

Deibert, Benjamin J.; Velasco, Ever; Liu, Wei; ...

2016-06-23

Here, we report the synthesis, structure, and photoluminescence properties of a new bismuth based luminescent metal-organic framework (LMOF). The framework is comprised of a 9-coordinated Bi 3+ building unit and 4', 4''', 4''''', 4'''''''-(ethene-1,1,2,2-tetrayl)tetrakis([1,1'-biphenyl]-4-carboxylic acid) (H 4tcbpe) organic linker, which has strong yellow aggregation induced emission (AIE). The structure can be viewed as two interpenetrated 4,4-anionic nets that are stabilized by K + ions forming one-dimensional helical inorganic chains by connecting bismuth nodes through shared oxygen bonds. The as-made LMOF has a bluish emission centered at 459 nm with an internal quantum yield of 57% when excited at 360 nm.more » The emission properties of the LMOF were found to be highly solvochromic with respect to DMF. Upon partial solvent removal, the framework undergoes significant red-shifting to a greenish emission centered at 500 nm. Complete removal of DMF results in additional red-shifting fluorescence coupled with structural changes. The resulting material has strong blue-excitable (455 nm) yellow emission centered at 553 nm, with a quantum yield of 74%, which is maintained after heating in air for 5 days at 90°C. This is the second highest quantum yield value for blue-excited yellow emission among all reported LMOFs.« less

High-Performance Blue-Excitable Yellow Phosphor Obtained from an Activated Solvochromic Bismuth-Fluorophore Metal–Organic Framework

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

Deibert, Benjamin J.; Velasco, Ever; Liu, Wei

Here, we report the synthesis, structure, and photoluminescence properties of a new bismuth based luminescent metal-organic framework (LMOF). The framework is comprised of a 9-coordinated Bi 3+ building unit and 4', 4''', 4''''', 4'''''''-(ethene-1,1,2,2-tetrayl)tetrakis([1,1'-biphenyl]-4-carboxylic acid) (H 4tcbpe) organic linker, which has strong yellow aggregation induced emission (AIE). The structure can be viewed as two interpenetrated 4,4-anionic nets that are stabilized by K + ions forming one-dimensional helical inorganic chains by connecting bismuth nodes through shared oxygen bonds. The as-made LMOF has a bluish emission centered at 459 nm with an internal quantum yield of 57% when excited at 360 nm.more » The emission properties of the LMOF were found to be highly solvochromic with respect to DMF. Upon partial solvent removal, the framework undergoes significant red-shifting to a greenish emission centered at 500 nm. Complete removal of DMF results in additional red-shifting fluorescence coupled with structural changes. The resulting material has strong blue-excitable (455 nm) yellow emission centered at 553 nm, with a quantum yield of 74%, which is maintained after heating in air for 5 days at 90°C. This is the second highest quantum yield value for blue-excited yellow emission among all reported LMOFs.« less

Characterization of dehydration-induced luminescence of kaolinite

NASA Technical Reports Server (NTRS)

Lahav, N.; Coyne, L.; Lawless, J. G.

1985-01-01

The dehydration-induced luminescence of a colloidal kaolinite is investigated experimentally, with particular attention given to the effect of various treatments on the luminescence characteristics. It is found that the total photon count of the emitted light is linearly related to the film thickness up to a thickness of 30 microns; mechanical stress in the form of grinding increases the photon output and produces extensive changes in the emission kinetics. A direct check of the emission wavelength dependence (by using color filters) indicates that roughly 75 percent of the emission occurs in the wavelength range below 410 nm. It is also found that incorporation of fluorescent molecules into the kaolinite paste increases the photon output and may indicate the transfer of ultraviolet photons to the fluorescent probe.

Autofluorescence of atmospheric bioaerosols - fluorescent biomolecules and potential interferences

NASA Astrophysics Data System (ADS)

Pöhlker, C.; Huffman, J. A.; Pöschl, U.

2012-01-01

Primary biological aerosol particles (PBAP) are an important subset of air particulate matter with a substantial contribution to the organic aerosol fraction and potentially strong effects on public health and climate. Recent progress has been made in PBAP quantification by utilizing real-time bioaerosol detectors based on the principle that specific organic molecules of biological origin such as proteins, coenzymes, cell wall compounds and pigments exhibit intrinsic fluorescence. The properties of many fluorophores have been well documented, but it is unclear which are most relevant for detection of atmospheric PBAP. The present study provides a systematic synthesis of literature data on potentially relevant biological fluorophores. We analyze and discuss their relative importance for the detection of fluorescent biological aerosol particles (FBAP) by online instrumentation for atmospheric measurements such as the ultraviolet aerodynamic particle sizer (UV-APS) or the wide issue bioaerosol sensor (WIBS). In addition, we provide new laboratory measurement data for selected compounds using bench-top fluorescence spectroscopy. Relevant biological materials were chosen for comparison with existing literature data and to fill in gaps of understanding. The excitation-emission matrices (EEM) exhibit pronounced peaks at excitation wavelengths of ~280 nm and ~360 nm, confirming the suitability of light sources used for online detection of FBAP. They also show, however, that valuable information is missed by instruments that do not record full emission spectra at multiple wavelengths of excitation, and co-occurrence of multiple fluorophores within a detected sample will likely confound detailed molecular analysis. Selected non-biological materials were also analyzed to assess their possible influence on FBAP detection and generally exhibit only low levels of background-corrected fluorescent emission. This study strengthens the hypothesis that ambient supermicron particle fluorescence in wavelength ranges used for most FBAP instruments is likely to be dominated by biological material and that such instrumentation is able to discriminate between FBAP and non-biological material in many situations. More detailed follow-up studies on single particle fluorescence are still required to reduce these uncertainties further, however.

Autofluorescence of atmospheric bioaerosols - fluorescent biomolecules and potential interferences

NASA Astrophysics Data System (ADS)

Pöhlker, C.; Huffman, J. A.; Pöschl, U.

2011-09-01

Primary biological aerosol particles (PBAP) are an important subset of air particulate matter with a substantial contribution to the organic aerosol fraction and potentially strong effects on public health and climate. Recent progress has been made in PBAP quantification by utilizing real-time bioaerosol detectors based on the principle that specific organic molecules of biological origin such as proteins, coenzymes, cell wall compounds and pigments exhibit intrinsic fluorescence. The properties of many fluorophores have been well documented, but it is unclear which are most relevant for detection of atmospheric PBAP. The present study provides a systematic synthesis of literature data on potentially relevant biological fluorophores. We analyze and discuss their relative importance for the detection of fluorescent biological aerosol particles (FBAP) by online instrumentation for atmospheric measurements such as the ultraviolet aerodynamic particle sizer (UV-APS) or the wide issue bioaerosol sensor (WIBS). In addition, we provide new laboratory measurement data for selected compounds using bench-top fluorescence spectroscopy. Relevant biological materials were chosen for comparison with existing literature data and to fill in gaps of understanding. The excitation-emission matrices (EEM) exhibit pronounced peaks at excitation wavelengths of ~280 nm and ~360 nm, confirming the suitability of light sources used for online detection of FBAP. They also show, however, that valuable information is missed by instruments that do not record full emission spectra at multiple wavelengths of excitation, and co-occurrence of multiple fluorophores within a detected sample will likely confound detailed molecular analysis. Selected non-biological materials were also analyzed to assess their possible influence on FBAP detection and generally exhibit only low levels of background-corrected fluorescent emission. This study strengthens the hypothesis that ambient supermicron particle fluorescence in wavelength ranges used for most FBAP instruments is likely to be dominated by biological material and that such instrumentation is able to discriminate between FBAP and non-biological material in many situations. More detailed follow-up studies on single particle fluorescence are still required to reduce these uncertainties further, however.

Rapid Optical Detection and Classification of Microbes in Suspicious Powders

DTIC Science & Technology

2018-06-01

per kilogram (J kg–1) [sievert (Sv)] * Specific details regarding the implementation of SI units may be viewed at http://www.bipm.org/en/si...amino acids, peptides, and proteins. Above about 340 nm fluorescence occurs from multi -ring organic materials and their variants. Therefore, to...emission wavelength, and fluorescence intensity, represented as iso-intensity contours in the EEM diagram. The two different fluorophors shown with

The photophysics of singlet, triplet, and degradation trap states in 4,4-N,N'-dicarbazolyl-1,1'-biphenyl

NASA Astrophysics Data System (ADS)

Jankus, Vygintas; Winscom, Chris; Monkman, Andrew P.

2009-02-01

In this paper we report the results of optical characterization of 4,4-N,N'-dicarbazolyl-1,1'-biphenyl (CBP), known as a host material for phosphorescent light emitting devices. Using absorption, steady state, and time-resolved spectroscopy, we explore the singlet and triplet states in solid and solution samples of CBP. In solutions we observe two distinct short-lived states with well-resolved emission originating from individual molecule singlet states (at 365 and 380 nm) and "quenching" low energy (LE) states (at 404 and 424 nm). The latter are seen only in saturated solutions and solid samples. Both of those species have different lifetimes. After UV exposure of very concentrated degassed solution the intensities of the LE bands starts to decrease. The longer the solution is exposed to UV, the less emission is seen at 404 and 424 nm, until it is totally gone. The spectrum of the highly concentrated solution is then the same as the spectrum of dilute solution, i.e., only emission at 365 and 380 nm is present. An increase in intensities of the singlet emission peaks correlates with an increase in UV exposure time. Similar behavior is observed in evaporated CBP film. We propose that this behavior is due to chemical instability of the weak N-C bonding of carbazolyl moiety—this creates new degradational species over time which dissociate after exposure to UV. We believe this to be the reason for variation in CBP fluorescence and delayed fluorescence spectra recorded by various research groups. Further, we detected two types of very long-lived states. One of these states (higher energy) is ascribed to molecular phosphorescence emission, the other to emission from low energy triplet trap states which we relate to degradational species. We propose that triplets are more easily caught by these latter sites when their hopping rate increases, and they emit inefficiently from these lower energy sites.

A single-photon fluorescence and multi-photon spectroscopic study of atherosclerotic lesions

NASA Astrophysics Data System (ADS)

Smith, Michael S. D.; Ko, Alex C. T.; Ridsdale, Andrew; Schattka, Bernie; Pegoraro, Adrian; Hewko, Mark D.; Shiomi, Masashi; Stolow, Albert; Sowa, Michael G.

2009-06-01

In this study we compare the single-photon autofluorescence and multi-photon emission spectra obtained from the luminal surface of healthy segments of artery with segments where there are early atherosclerotic lesions. Arterial tissue was harvested from atherosclerosis-prone WHHL-MI rabbits (Watanabe heritable hyperlipidemic rabbit-myocardial infarction), an animal model which mimics spontaneous myocardial infarction in humans. Single photon fluorescence emission spectra of samples were acquired using a simple spectrofluorometer set-up with 400 nm excitation. Samples were also investigated using a home built multi-photon microscope based on a Ti:sapphire femto-second oscillator. The excitation wavelength was set at 800 nm with a ~100 femto-second pulse width. Epi-multi-photon spectroscopic signals were collected through a fibre-optics coupled spectrometer. While the single-photon fluorescence spectra of atherosclerotic lesions show minimal spectroscopic difference from those of healthy arterial tissue, the multi-photon spectra collected from atherosclerotic lesions show marked changes in the relative intensity of two-photon excited fluorescence (TPEF) and second-harmonic generation (SHG) signals when compared with those from healthy arterial tissue. The observed sharp increase of the relative SHG signal intensity in a plaque is in agreement with the known pathology of early lesions which have increased collagen content.

Absorption and Fluorescence Properties of Chromophoric Dissolved Organic Matter Produced by Algae.

PubMed

Peng, Tong; Lu, Xiao-lan; Su, Rong-guo; Zhang, Dong-mei

2015-09-01

Four kinds of diatom (Chaetoceros curvisetus, Phaeodactylum tricornutum, Nitzschia closterium f. minutissima and Navicula halophile) and two kinds of dinoflagellates (Prorocentrum donghaiense and Gymnodinium) were cultured under laboratory conditions. Variations of optical properties of chromophoric dissolved organic matter (CDOM) were studied with absorption and fluorescence excitation-emission matrix spectroscopy(EEM) during growth of marine microalgae in incubation experiment. Absorption spectrum revealed absorption coefficient a(355) (CDOM absorption coefficients at 355 nm) of 6 kinds of marine microalgae above increased by 64.8%, 242.3%, 535.1%, 903.2%, 836% and 196.4%, respectively. Simultaneously, the absorption spectral slope (Sg), determined between 270 and 350 nm, representing the size of molecular weight of CDOM and humic-like composition, decreased by 8.7%, 34.6%, 39.4%, 53.1%, 46.7%, and 35.7%, respectively. Applying parallel factor analysis (PARAFAC) together with EEM got four components of CDOM: C1(Ex/Em=350(260) nm/450 nm), C2 (Ex/Em=260(430) nm/525 nm), C3 (Ex/Em=325 nm/400 nm) and C4(Ex/Em=275 nm/325 nm), which were relative to three humic-like and one protein-like fluorescent components of Nitzschia closterium f. minutissima and Navicula halophile. In incubation experiment, fluorescence intensity of these four components during growth of Nitzschia closterium f. minutissima increased by, respectively, 8.68, 24.9, 7.19 and 39.8 times, and those of Navicula halophile increased by 2.64, 0.07, 4.39 and 12.4 times, respectively. Significant relationships were found between the fluorescence intensity of four components of CDOM, a(355) and Sg. All results demonstrated that both content and molecular weight of CDOM produced by diatom and dinoflagellate studied in incubation experiment increased, but these two parameters changed more obviously of the diatom than those of dinoflagellate; the proportion of humic-like components in the composition of CDOM also increased clearly with the growth of marine microalgae, but protein-like fluorescent component had only a slow growth. Furthermore, the absorption spectrum of CDOM produced by different species of algae changed obviously and the relative composition fluorescence intensity of CDOM produced by different microalgae were found to vary among different composition from EEM, which suggested CDOM produced by different microalgae make quite different contributions to CDOM in natural seawater.

Crystallization-induced emission enhancement: A novel fluorescent Au-Ag bimetallic nanocluster with precise atomic structure

PubMed Central

Chen, Tao; Yang, Sha; Chai, Jinsong; Song, Yongbo; Fan, Jiqiang; Rao, Bo; Sheng, Hongting; Yu, Haizhu; Zhu, Manzhou

2017-01-01

We report the first noble metal nanocluster with a formula of Au4Ag13(DPPM)3(SR)9 exhibiting crystallization-induced emission enhancement (CIEE), where DPPM denotes bis(diphenylphosphino)methane and HSR denotes 2,5-dimethylbenzenethiol. The precise atomic structure is determined by x-ray crystallography. The crystalline state of Au4Ag13 shows strong luminescence at 695 nm, in striking contrast to the weak emission of the amorphous state and hardly any emission in solution phase. The structural analysis and the density functional theory calculations imply that the compact C–H⋯π interactions significantly restrict the intramolecular rotations and vibrations and thus considerably enhance the radiative transitions in the crystalline state. Because the noncovalent interactions can be easily modulated via varying the chemical environments, the CIEE phenomenon might represent a general strategy to amplify the fluorescence from weakly (or even non-) emissive nanoclusters. PMID:28835926

Preparation of carbon quantum dots from cigarette filters and its application for fluorescence detection of Sudan I.

PubMed

Anmei, Su; Qingmei, Zhong; Yuye, Chen; Yilin, Wang

2018-09-06

Carbon quantum dots (CQDs) with quantum yield of 14% were successfully synthesized via a simple, low-cost, and green hydrothermal treatment using cigarette filters as carbon source for the first time. The obtained CQDs showed a strong emission at the wavelength of 465 nm, with an optimum excitation of 365 nm.Sudan I with maximum absorption wavelength at 477 nm could selectively quench the fluorescence of CQDs. Based on this principle, a fluorescence probe was developed for Sudan I determination. Furthermore, the quenching mechanism of the CQDs was elucidated. A linear relationship was found in the range of 2.40-104.0 μmol/L Sudan I with the detection limit (3σ/k) of 0.95 μmol/L. Satisfactory results were achieved when the method was submitted to the determination of Sudan I in food samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Influence of functional groups on the photophysical properties of dimethylamino chalcones as laser dyes

NASA Astrophysics Data System (ADS)

Ibnaouf, K. H.; Elzupir, A. O.; AlSalhi, M. S.; Alaamer, Abdulaziz S.

2018-02-01

In this report, a series of 3-(4-(Dimethylamino) phenyl)-1-(4,3 di-substituted phenyl)-(2E) - propen -1-one chalcones was synthesised and examined as optical materials. The influence of functional groups (FG) and solvents on their photophysical properties was investigated. These include absorption, fluorescence, Stokes shift, and amplified spontaneous emission (ASE). The absorption spectra of all compounds showed a wavelength band in the range 404-429 nm, whereas the fluorescence spectra exhibited a band at 470-535 nm. We found that the fluorescence intensity was inversely proportional to the concentration of chalcones. The FGs and solvents had an amazing effect on the photophysical properties of the synthesised materials. Unexpectedly, the electron withdrawing group showed a highly red shift, whereas the electron donating group exhibited a blue shift. Further, these compounds showed large Stokes shifts (up to 5800 cm-1). ASE was observed under pump pulse laser excitation, and the wavelengths were tuned from 509 to 566 nm.

Dye-Enhanced Multimodal Confocal Imaging of Brain Cancers

NASA Astrophysics Data System (ADS)

Wirth, Dennis; Snuderl, Matija; Sheth, Sameer; Curry, William; Yaroslavsky, Anna

2011-04-01

Background and Significance: Accurate high resolution intraoperative detection of brain tumors may result in improved patient survival and better quality of life. The goal of this study was to evaluate dye enhanced multimodal confocal imaging for discriminating normal and cancerous brain tissue. Materials and Methods: Fresh thick brain specimens were obtained from the surgeries. Normal and cancer tissues were investigated. Samples were stained in methylene blue and imaged. Reflectance and fluorescence signals were excited at 658nm. Fluorescence emission and polarization were registered from 670 nm to 710 nm. The system provided lateral resolution of 0.6 μm and axial resolution of 7 μm. Normal and cancer specimens exhibited distinctively different characteristics. H&E histopathology was processed from each imaged sample. Results and Conclusions: The analysis of normal and cancerous tissues indicated clear differences in appearance in both the reflectance and fluorescence responses. These results confirm the feasibility of multimodal confocal imaging for intraoperative detection of small cancer nests and cells.

White organic light-emitting diodes based on doped and ultrathin Rubrene layer

NASA Astrophysics Data System (ADS)

Li, Yi; Jiang, Yadong; Wen, Wen; Yu, Junsheng

2010-10-01

Based on a yellow fluorescent dye of 5, 6, 11, 12-tetraphenylnaphthacene (Rubrene), WOLEDs were fabricated, with doping structure and ultrathin layer structure utilized in the devices. By doping Rubrene into blue-emitting N,N'-bis-(1- naphthyl)-N,N'-biphenyl-1,1'-biphenyl-4,4'-diamine (NPB), the device with a structure of indium-tin-oxide (ITO)/NPB (40 nm)/NPB:Rubrene (0.25 wt%, 7 nm)/2,9-dimethyl-4,7-diphenyl-1,10-phenanthroline (BCP) (30 nm)/Mg:Ag exhibited a warm white light with Commissions Internationale De L'Eclairage (CIE) coordinates of (0.38, 0.41) at 12 V. The electroluminescent spectrum of the OLED consisted of blue and yellow fluorescent emissions, the intensity of blue emission increased gradually relative to the orange emission with increasing voltage. This is mainly due to the recombination zone shifted towards the anode side as the transmission rate of electrons grows faster than that of holes under higher bias voltage. A maximum luminance of 7300 cd/m2 and a maximum power efficiency of 0.57 lm/W were achieved. Comparatively, by utilizing ultrathin dopant layer, the device with a structure of ITO/NPB (40 nm)/Rubrene (0.3 nm)/NPB (7 nm)/BCP (30 nm)/Mg:Ag achieved a low turn-on voltage of 3 V and a more stable white light. The peaks of EL spectra located at 430 and 560 nm corresponding to the CIE coordinates of (0.32, 0.32) under bias voltage ranging from 5 to 15 V. A maximum luminance of 5630 cd/m2 and a maximum power efficiency of 0.6 lm/W were achieved. The balanced spectra were attributed to the stable confining of charge carriers and exciton by the thin emitting layers. Hence, with simple device structure and fabricating process, the device with ultrathin layer achieved low turn-on voltage, stable white light emitting and higher power efficiency.

Spatial variability of oceanic phycoerythrin spectral types derived from airborne laser-induced fluorescence emissions

NASA Astrophysics Data System (ADS)

Hoge, Frank E.; Wright, C. Wayne; Kana, Todd M.; Swift, Robert N.; Yungel, James K.

1998-07-01

We report spatial variability of oceanic phycoerythrin spectral types detected by means of a blue spectral shift in airborne laser-induced fluorescence emission. The blue shift of the phycoerythrobilin fluorescence is known from laboratory studies to be induced by phycourobilin chromophore substitution at phycoerythrobilin chromophore sites in some strains of phycoerythrin-containing marine cyanobacteria. The airborne 532-nm laser-induced phycoerythrin fluorescence of the upper oceanic volume showed distinct segregation of cyanobacterial chromophore types in a flight transect from coastal water to the Sargasso Sea in the western North Atlantic. High phycourobilin levels were restricted to the oceanic (oligotrophic) end of the flight transect, in agreement with historical ship findings. These remotely observed phycoerythrin spectral fluorescence shifts have the potential to permit rapid, wide-area studies of the spatial variability of spectrally distinct cyanobacteria, especially across interfacial regions of coastal and oceanic water masses. Airborne laser-induced phytoplankton spectral fluorescence observations also further the development of satellite algorithms for passive detection of phytoplankton pigments. Optical modifications to the NASA Airborne Oceanographic Lidar are briefly described that permitted observation of the fluorescence spectral shifts.

Long wavelength fluorescence based biosensors for in vivo continuous monitoring of metabolites

NASA Astrophysics Data System (ADS)

Thomas, Joseph; Ambroise, Arounaguiry; Birchfield, Kara; Cai, Wensheng; Sandmann, Christian; Singh, Sarabjit; Weidemaier, Kristin; Pitner, J. Bruce

2006-02-01

The early stage development studies of novel implantable continuous metabolite sensor systems for glucose, lactate and fatty acids are discussed. These sensors utilize non-enzymatic "reagentless" sensor systems based on NIR fluorophore-labeled binding proteins. For in vivo applications, NIR fluorescence based systems (beyond 600 nm) have the added benefit of reduced interference from background scattering, tissue and serum absorption and cell auto-fluorescence. The long wavelength emission facilitates implanted sensor disks to transmit fluorescence to an external reader through wireless connections and the resulting fluorescence signals can be correlated to metabolite concentrations. We have developed a prototype optical system that uses a bifurcated optical fiber to transmit excitation and read emission at the surface of the skin. With this system, fluorescence signals were read over time through animal skin. The changes in glucose concentration were studied using immobilized sensor proteins and were compared to non-immobilized sensors in solution. For sensors in solution, no response delay was observed. For immobilized systems, the fluorescence response showed a delay corresponding to the diffusion time for the metabolite to equilibrate within the sensor.

Copper nanoclusters as probes for turn-on fluorescence sensing of L-lysine.

PubMed

Zhang, Mingming; Qiao, Juan; Zhang, Shufeng; Qi, Li

2018-05-15

Herein, a unique protocol based on copper nanoclusters (CuNCs) probe for turn-on fluorescence sensing of L-lysine was developed. The fluorescent CuNCs with ovalbumin as the stabilizer was prepared by a simple, one-step and green method. When 370 nm was used as the excitation wavelength, the resultant CuNCs exhibited a pale blue fluorescence with the maximum emission at 440 nm. Interestingly, existence of L-lysine evoked the obvious fluorescence intensity increase of CuNCs. The detection limit of the proposed method for L-lysine was 5.5 μM, with a good linear range from 10.0 μM to 1.0 mM (r 2 = 0.999). Moreover, the possible mechanism for enhanced fluorescence intensity of CuNCs by addition of L-lysine was explored and discussed briefly. Further, the as-prepared fluorescent CuNCs was successfully applied in detection of L-lysine in urine. Our results demonstrated that L-lysine could be monitored by the probe, providing new path for construction of CuNCs as fluorescent probes and showing great potential in quantification of L-lysine in real samples. Copyright © 2018 Elsevier B.V. All rights reserved.

Benzofurazan Sulfides for Thiol Imaging and Quantification in Live Cells through Fluorescence Microscopy

PubMed Central

Li, Yinghong; Yang, Yang; Guan, Xiangming

2012-01-01

Thiol groups play a significant role in various cellular functions. Cellular thiol concentrations can be affected by various physiological or pathological factors. A fluorescence imaging agent that can effectively and specifically image thiols in live cells through fluorescence microscopy is desirable for live cell thiol monitoring. Benzofurazan sulfides 1a–e were synthesized and found to be thiol specific fluorogenic agents except 1d. They are not fluorescent but form strong fluorescent thiol adducts after reacting with thiols through a sulfide-thiol exchange reaction. On the other hand, they exhibit no reaction with other biologically relevant nucleophilic functional groups such as -NH2, -OH, or -COOH revealing the specificity for the detection of thiols. Sulfide 1a was selected to confirm its ability to image cellular thiols through fluorescence microscopy. The compound was demonstrated to effectively image and quantify thiol changes in live cells through fluorescence microscopy using 430 nm and 520 nm as the excitation and emission wavelengths respectively. The quantification results of total thiol in live cells obtained from fluorescence microscopy were validated by an HPLC/UV total thiol assay method. The reagents and method will be of a great value to thiol redox-related research. PMID:22794193

Photo diagnosis of early pre cancer (LSIL) in genital tissue

NASA Astrophysics Data System (ADS)

Vaitkuviene, A.; Andersen-Engels, S.; Auksorius, E.; Bendsoe, N.; Gavriushin, V.; Gustafsson, U.; Oyama, J.; Palsson, S.; Soto Thompson, M.; Stenram, U.; Svanberg, K.; Viliunas, V.; De Weert, M. J.

2005-11-01

Permanent infections recognized as oncogenic factor. STD is common concomitant diseases in early precancerous genital tract lesions. Simple optical detection of early regressive pre cancer in cervix is the aim of this study. Hereditary immunosupression most likely is risk factor for cervical cancer development. Light induced fluorescence point monitoring fitted to live cervical tissue diagnostics in 42 patients. Human papilloma virus DNR in cervix tested by means of Hybrid Capture II method. Ultraviolet (337 nm) laser excited fluorescence spectra in the live cervical tissue analyzed by Principal Component (PrC) regression method and spectra decomposition method. PCr method best discriminated pathology group "CIN I and inflammation"(AUC=75%) related to fluorescence emission in short wave region. Spectra decomposition method suggested a few possible fluorophores in a long wave region. Ultraviolet (398 nm) light excitation of live cervix proved sharp selective spectra intensity enhancement in region above 600nm for High-grade cervical lesion. Conclusion: PC analysis of UV (337 nm) light excitation fluorescence spectra gives opportunity to obtain local immunity and Low-grade cervical lesion related information. Addition of shorter and longer wavelengths is promising for multi wave LIF point monitoring method progress in cervical pre-cancer diagnostics and utility for cancer prevention especially in developing countries.

Visible-to-NIR Electrochromic Device Prepared from a Thermally Polymerizable Electroactive Organic Monomer.

PubMed

Wałęsa-Chorab, Monika; Skene, W G

2017-06-28

A monomer (1) consisting of a benzothiadiazole core flanked by two triphenylamines and two styrene pendant moieties was prepared. The monomer was fluorescent with its emission spanning 145 nm in the visible, contingent on the organic solvent used for the measurement. In addition to its positive solvatochromism, the absolute fluorescence quantum yield (Φ fl ) was consistently >20% with values >80% being measured in hexane, toluene, diethyl ether, and toluene. 1 could be reversibly oxidized with an oxidation potential of 880 mV vs SCE. The monomer could be immobilized on ITO-coated glass substrates. The resulting 425 nm thick immobilized film (poly-1) was 15% thinner than the monomer coating deposited by spray- and spin-coating. The electroactive film did not delaminate from the electrode upon either washing or cycling electrochemically between its oxidized and neutral states. Its absorption at 460 nm bleached upon electrochemical oxidation with the formation of a strong absorption at 880 nm and in the NIR, similar to 1. The perceived reversible color change with applied potential switched between yellow and gray. The fluorescence intensity of poly-1 could be switched with applied potentials. A passive transmissive device prepared from poly-1 was both electrochromic and fluorochromic, exhibiting reversible color change and fluorescence quenching.

Two-photon excited fluorescence spectroscopy and imaging of melanin in vitro and in vivo

NASA Astrophysics Data System (ADS)

Krasieva, Tatiana B.; Liu, Feng; Sun, Chung-Ho; Kong, Yu; Balu, Mihaela; Meyskens, Frank L.; Tromberg, Bruce J.

2012-03-01

The ability to detect early melanoma non-invasively would improve clinical outcome and reduce mortality. Recent advances in two-photon excited fluorescence (TPEF) in vivo microscopy offer a powerful tool in early malignant melanoma diagnostics. The goal of this work was to develop a TPEF optical index for measuring relative concentrations of eumelanin and pheomelanin since ex vivo studies show that changes in this ratio have been associated with malignant transformation. We acquired TPEF emission spectra (λex=1000 nm) of melanin from several specimens, including human hair, malignant melanoma cell lines, and normal melanocytes and keratinocytes in different skin layers (epidermis, papillary dermis) in five healthy volunteers in vivo. We found that the pheomelanin emission peaks at around 620 nm and is blue-shifted from the eumelanin with broad maximum at 640-680nm. We defined "optical melanin index" (OMI) as a ratio of fluorescence signal intensities measured at 645 nm and 615nm. The measured OMI for a melanoma cell line MNT-1 was 1.6+/-0.2. The MNT-46 and MNT-62 lines (Mc1R gene knockdown) showed an anticipated change in melanins production ratio and had OMI of 0.55+/-0.05 and 0.17+/-0.02, respectively, which strongly correlated with HPLC data obtained for these lines. Average OMI measured for basal cells layers (melanocytes and keratinocytes) in normal human skin type I, II-III (not tanned and tanned) in vivo was 0.5, 1.05 and 1.16 respectively. We could not dependably detect the presence of pheomelanin in highly pigmented skin type V-VI. These data suggest that a non-invasive TPEF index could potentially be used for rapid melanin ratio characterization both in vitro and in vivo, including pigmented lesions.

Enhanced emission of nile red fluorescent nanoparticles embedded in hybrid sol-gel glasses.

PubMed

Ferrer, Maria L; del Monte, Francisco

2005-01-13

Highly fluorescent Nile Red (NR) nanoparticles embedded in a hybrid sol-gel glass are reported. The crystallite growth within the confined system created by the porous hybrid matrix results in NR nanoparticles of averaged dimensions below 36 nm. The preparation process allows for the control of both the conformation adopted by single NR molecules prior to aggregation (e.g., near planar) and the configuration of the aggregates (e.g., oblique with phi < 54.7 degrees) prior to their assembly in the supramolecular architecture which ultimately forms the nanoparticles. The full preservation of the fluorescent configuration of the aggregates in the nanoparticles is confirmed through the application of the exciton theory, and it is responsible for the significant increase of the fluorescence emission intensity (e.g., up to 525- and 70-fold as compared to that obtained for single NR molecules embedded in pure and hybrid silica glasses, respectively).

Potential applications of near infrared auto-fluorescence spectral polarized imaging for assessment of food quality

NASA Astrophysics Data System (ADS)

Zhou, Kenneth J.; Chen, Jun

2016-03-01

The current growing of food industry for low production costs and high efficiency needs for maintenance of high-quality standards and assurance of food safety while avoiding liability issues. Quality and safety of food depend on physical (texture, color, tenderness etc.), chemical (fat content, moisture, protein content, pH, etc.), and biological (total bacterial count etc.) features. There is a need for a rapid (less than a few minutes) and accurate detection system in order to optimize quality and assure safety of food. However, the fluorescence ranges for known fluorophores are limited to ultraviolet emission bands, which are not in the tissue near infrared (NIR) "optical window". Biological tissues excited by far-red or NIR light would exhibit strong emission in spectral range of 650-1,100 nm although no characteristic peaks show the emission from which known fluorophores. The characteristics of the auto-fluorescence emission of different types of tissues were found to be different between different tissue components such as fat, high quality muscle food. In this paper, NIR auto-fluorescence emission from different types of muscle food and fat was measured. The differences of fluorescence intensities of the different types of muscle food and fat emissions were observed. These can be explained by the change of the microscopic structure of physical, chemical, and biological features in meat. The difference of emission intensities of fat and lean meat tissues was applied to monitor food quality and safety using spectral polarized imaging, which can be detect deep depth fat under the muscle food up to several centimeter.

A long-wavelength fluorescent squarylium cyanine dye possessing boronic acid for sensing monosaccharides and glycoproteins with high enhancement in aqueous solution.

PubMed

Saito, Shingo; Massie, Tara L; Maeda, Takeshi; Nakazumi, Hiroyuki; Colyer, Christa L

2012-01-01

Fluorescence sensing of saccharides and glycoproteins using a boronic acid functionalized squarylium cyanine dye ("SQ-BA") is characterized in terms of synthetic, fluorometric, thermodynamic and kinetic parameters. In our previous work, this newly synthesized dye was successfully applied to the separation and quantification of Gram-positive bacteria by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF); however, the fundamental properties of the dye and its saccharide complexes still required elucidation, as presented in this paper. The dye itself forms nonemissive, soluble aggregates in aqueous solution. With the addition of a monosaccharide, the dye aggregate dissociates to form an emissive monomer accompanied by the formation of a cyclic cis-diol ester with long-wavelength emission (λ(ex) = 630 nm, λ(em) = 660 nm). A very large fluorescence enhancement factor of 18× was observed for the sensing dye as a fructose complex at pH 10, yielding a limit of detection of 10 μM fructose. The relative order of fluorescence enhancement of SQ-BA with other monosaccharides was found to be: fructose > ribose > arabinose ≈ galactose > xylose > mannose > rhamnose > fucose ≈ glucose; and apparent affinity constants of 10(2.80), 10(2.08) and 10(0.86) M(-1) were determined for fructose, ribose and glucose, respectively. Formation of the emissive complexes occurred within minutes, proving the kinetics of the sugar-dye interactions to be suitable for on-column labeling methods in CE-LIF. Furthermore, the sensing dye was successfully applied to glycoproteins, mucin type I-S and type III, which were detected with high sensitivity in batch aqueous solution as a result of the sugar-selective boronic acid-diol esterification as well as hydrophobic interactions.

A Long-Wavelength Fluorescent Squarylium Cyanine Dye Possessing Boronic Acid for Sensing Monosaccharides and Glycoproteins with High Enhancement in Aqueous Solution

PubMed Central

Saito, Shingo; Massie, Tara L.; Maeda, Takeshi; Nakazumi, Hiroyuki; Colyer, Christa L.

2012-01-01

Fluorescence sensing of saccharides and glycoproteins using a boronic acid functionalized squarylium cyanine dye (“SQ-BA”) is characterized in terms of synthetic, fluorometric, thermodynamic and kinetic parameters. In our previous work, this newly synthesized dye was successfully applied to the separation and quantification of Gram-positive bacteria by capillary electrophoresis with laser-induced fluorescence detection (CE-LIF); however, the fundamental properties of the dye and its saccharide complexes still required elucidation, as presented in this paper. The dye itself forms nonemissive, soluble aggregates in aqueous solution. With the addition of a monosaccharide, the dye aggregate dissociates to form an emissive monomer accompanied by the formation of a cyclic cis-diol ester with long-wavelength emission (λex = 630 nm, λem = 660 nm). A very large fluorescence enhancement factor of 18× was observed for the sensing dye as a fructose complex at pH 10, yielding a limit of detection of 10 μM fructose. The relative order of fluorescence enhancement of SQ-BA with other monosaccharides was found to be: fructose > ribose > arabinose ≈ galactose > xylose > mannose > rhamnose > fucose ≈ glucose; and apparent affinity constants of 102.80, 102.08 and 100.86 M−1 were determined for fructose, ribose and glucose, respectively. Formation of the emissive complexes occurred within minutes, proving the kinetics of the sugar-dye interactions to be suitable for on-column labeling methods in CE-LIF. Furthermore, the sensing dye was successfully applied to glycoproteins, mucin type I–S and type III, which were detected with high sensitivity in batch aqueous solution as a result of the sugar-selective boronic acid-diol esterification as well as hydrophobic interactions. PMID:22778592