Velocity landscape correlation resolves multiple flowing protein populations from fluorescence image time series.

Science.gov (United States)

Pandžić, Elvis; Abu-Arish, Asmahan; Whan, Renee M; Hanrahan, John W; Wiseman, Paul W

2018-02-16

Molecular, vesicular and organellar flows are of fundamental importance for the delivery of nutrients and essential components used in cellular functions such as motility and division. With recent advances in fluorescence/super-resolution microscopy modalities we can resolve the movements of these objects at higher spatio-temporal resolutions and with better sensitivity. Previously, spatio-temporal image correlation spectroscopy has been applied to map molecular flows by correlation analysis of fluorescence fluctuations in image series. However, an underlying assumption of this approach is that the sampled time windows contain one dominant flowing component. Although this was true for most of the cases analyzed earlier, in some situations two or more different flowing populations can be present in the same spatio-temporal window. We introduce an approach, termed velocity landscape correlation (VLC), which detects and extracts multiple flow components present in a sampled image region via an extension of the correlation analysis of fluorescence intensity fluctuations. First we demonstrate theoretically how this approach works, test the performance of the method with a range of computer simulated image series with varying flow dynamics. Finally we apply VLC to study variable fluxing of STIM1 proteins on microtubules connected to the plasma membrane of Cystic Fibrosis Bronchial Epithelial (CFBE) cells. Copyright © 2018 Elsevier Inc. All rights reserved.

Variation in U.V. primary fluorescence-intensity of vital cells depending on 60Co γ-radiation dose

International Nuclear Information System (INIS)

Merkle, K.

1978-01-01

Using impulse-cytofluorophotometry in the ultra-violet spectral region it has been shown on vital, unstained Ehrlich ascites tumour cells that the primary fluorescence intensity of this tumour was on day 11 after transplantation 20 per cent higher than on day 8. Storage of the vital cells for 25 min at 20 0 C had no effect on this result. When the cells were exposed to 60 Co γ-radiation on day 6, a new stable fluorescence level was established after 20 hours. Measurements of the primary fluorescence intensity depending on dose have shown a significant rise starting from 75 rad at 48 hours after irradiation. The fluorescence intensity rose by 42.5 per cent of the control value at 3000 rad, but only by 31.5 per cent on exposure to 4000 rad. (author)

Study on silicon oxide coated on silver nanocrystal to enhance fluorescence intensity of rare earth complexes

Energy Technology Data Exchange (ETDEWEB)

Qu, Yan-rong; Lin, Xue-mei; Wang, Ai-ling; Wang, Zhong-xia; Kang, Jie; Chu, Hai-bin, E-mail: binghai99@gmail.com; Zhao, Yong-liang, E-mail: hxzhaoyl@163.com

2014-10-15

Twelve kinds of rare earth complexes were synthesized using halo-benzoic acid as anion ligand and Sm{sup 3+} and Dy{sup 3+} as central ions, respectively. The complexes were characterized by elemental analysis, rare earth coordination titration and electrospray ionization mass spectra, from which the compositions of the complexes were confirmed to be RE(p-FBA){sub 3}·H{sub 2}O, RE(p-ClBA){sub 3}·2H{sub 2}O, RE(p-BrBA){sub 3}·H{sub 2}O, RE(o-FBA){sub 3}·2H{sub 2}O, RE(o-ClBA){sub 3}·H{sub 2}O, RE(o-BrBA){sub 3}·H{sub 2}O (RE=Sm{sup 3+}, Dy{sup 3+}). Besides, IR spectra and UV–visible absorption spectroscopy indicated that the carboxyl oxygen atoms of ligands coordinated to the rare earth ions. Moreover, Ag@SiO{sub 2} core–shell nanoparticles (NPs) were prepared via a modified Stöber method. The average diameters of silver cores were typically between 60 nm and 70 nm, and the thicknesses of the SiO{sub 2} shells were around 10 nm, 15 nm and 25 nm, respectively. The influence of Ag@SiO{sub 2} NPs on the luminescence properties of the rare earth complexes showed that the luminescence intensities of rare earth complexes were enhanced remarkably. As the thickness of SiO{sub 2} shell increases in the range of 10–25 nm, the effect of metal-enhanced fluorescence become obvious. The mechanism of the changes of the fluorescence intensity is also discussed. - Highlights: • Among 10–25 nm, the thicker the shell thickness, the better the fluorescence effect. • The strong the intensity of the pure complexes, the smaller the multiple enhanced. • The intensity of Sm(p-BrBA){sub 3}·H{sub 2}O is the strongest among Sm(p-XBA){sub 3}·nH{sub 2}O complexes. • The intensity of Dy(p-ClBA){sub 3}·2H{sub 2}O is the strongest among Dy(p-XBA){sub 3}·nH{sub 2}O complexes. • When halogen is in o-position, the intensity of RE(o-ClBA){sub 3}·H{sub 2}O is the strongest.

Sub-annual paleoenvironmental information evaluated from intensity variations of fluorescent annual layers in a stalagmite from Ryuo-do Cave, Nagasaki Prefecture, western Japan

Science.gov (United States)

Sasaki, Hana; Onishi, Yuri; Ishihara, Yoshiro; Yoshimura, Kazuhisa

2017-04-01

vary stratigraphically, and multiple types of fluorescence intensity pattern are observed in the stalagmite. When the co-precipitation of FA is governed by the hiatus model, it is suggested that a gradual increase in the annual layers will result from a large accumulation of calcite after the annual peak in the FA concentration, whereas there will be a gradual decrease if the main growth occurs before the annual peak in FA concentration. However, in the case of the PC model, a gradually increasing type of pattern is formed if the main growth occurs before the annual peak in FA concentration, and a gradually decreasing type is formed if the main growth occurs afterwards. If the annual peak of FA concentration occurs several months after high summer, it is suggested that intervals showing a gradually increasing type were formed in winter, and intervals showing a gradually decreasing type were formed in the early summer, in the case of the hiatus model. In the case of PC model, the seasons are reversed. In the climatic environment around the Ryuo-do Cave, the growth rates of stalagmites are affected by cave air circulation in winter and by rainfall (rainy season) in early summer.

Enhancing fluorescence intensity of Ellagic acid in Borax-HCl-CTAB micelles

Science.gov (United States)

Wang, Feng; Huang, Wei; Zhang, Shuai; Liu, Guokui; Li, Kexiang; Tang, Bo

2011-03-01

Ellagic acid (C 14H 6O 8), a naturally occurring phytochemical, found mainly in berries and some nuts, has anticarcinogenic and antioxidant properties. It is found that fluorescence of Ellagic acid (EA) is greatly enhanced by micelle of cetyltrimethylammonium bromide (CTAB) surfactant. Based on this effect, a sensitive proposed fluorimetric method was applied for the determination of Ellagic acid in aqueous solution. In the Borax-HCl buffer, the fluorescence intensity of Ellagic acid in the presence of CTAB is proportional to the concentration of Ellagic acid in range from 8.0 × 10 -10 to 4.0 × 10 -5 mol L -1; and the detection limits are 3.2 × 10 -10 mol L -1 and 5.9 × 10 -10 mol L -1 excited at 266 nm and 388 nm, respectively. The actual samples of pomegranate rinds are simply manipulated and satisfactorily determined. The interaction mechanism studies argue that the negative EA-Borax complex is formed and solubilized in the cationic surfactant CTAB micelle in this system. The fluorescence intensity of EA enhances because the CTAB micelle provides a hydrophobic microenvironment for EA-Borax complex, which can prevent collision with water molecules and decrease the energy loss of EA-Borax complex.

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

International Nuclear Information System (INIS)

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

2004-01-01

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

Multiple X-ray tomography using transmitted, scattered and fluorescent radiation

International Nuclear Information System (INIS)

Cesareo, R.; Brunetti, A.; Golosio, B.; Lopes, R.T.; Barroso, R.C.; Donativi, M.; Castellano, A.; Quarta, S.

2003-01-01

A multiple CT-scanner is described, which contemporaneously uses transmitted, scattered and fluorescent X-rays for Imaging. The scanner is characterized by a small size X-ray tube and by four detectors: a ''pencil'' X-ray NaI(Tl) for transmitted tomography, a larger size NaI(Tl) for 90 C o Compton tomography, a thermoelectrically cooled Si-PIN or CdZnTe for fluorescent imaging and a CdZnTe for Rayleigh (or diffraction) tomography. Examples of applications are shown

Molecular Viscosity Sensors with Two Rotators for Optimizing the Fluorescence Intensity-Contrast Trade-Off.

Science.gov (United States)

Lee, Seung-Chul; Lee, Chang-Lyoul; Heo, Jeongyun; Jeong, Chan-Uk; Lee, Gyeong-Hui; Kim, Sehoon; Yoon, Woojin; Yun, Hoseop; Park, Sung O; Kwak, Sang Kyu; Park, Sung-Ha; Kwon, O-Pil

2018-02-26

A series of fluorescent molecular rotors obtained by introducing two rotational groups ("rotators"), which exhibit different rotational and electron-donating abilities, are discussed. Whereas the control molecular rotor, PH, includes a single rotator (the widely used phenyl group), the PO molecular rotors consist of two rotators (a phenyl group and an alkoxy group), which exhibit simultaneous strongly electron-donating and easy rotational abilities. Compared with the control rotor PH, PO molecular rotors exhibited one order of magnitude higher quantum yield (fluorescence intensity) and simultaneously exhibited significantly higher fluorescence contrast. These properties are directly related to the strong electron-donating ability and low energy barrier of rotation of the alkoxy group, as confirmed by dynamic fluorescence experiments and quantum chemical calculations. The PO molecular rotors exhibited two fluorescence relaxation pathways, whereas the PH molecular rotor exhibited a single fluorescence relaxation pathway. Cellular fluorescence imaging with PO molecular rotors for mapping cellular viscosity was successfully demonstrated. © 2018 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

High Intensity Exercise in Multiple Sclerosis

DEFF Research Database (Denmark)

Wens, Inez; Dalgas, Ulrik; Vandenabeele, Frank

2015-01-01

Introduction Low-to-moderate intensity exercise improves muscle contractile properties and endurance capacity in multiple sclerosis (MS). The impact of high intensity exercise remains unknown. Methods Thirty-four MS patients were randomized into a sedentary control group (SED, n = 11) and 2...... exercise groups that performed 12 weeks of a high intensity interval (HITR, n = 12) or high intensity continuous cardiovascular training (HCTR, n = 11), both in combination with resistance training. M.vastus lateralis fiber cross sectional area (CSA) and proportion, knee-flexor/extensor strength, body...... composition, maximal endurance capacity and self-reported physical activity levels were assessed before and after 12 weeks. Results Compared to SED, 12 weeks of high intensity exercise increased mean fiber CSA (HITR: +21±7%, HCTR: +23±5%). Furthermore, fiber type I CSA increased in HCTR (+29±6%), whereas type...

Chair-side detection of Prevotella Intermedia in mature dental plaque by its fluorescence.

Science.gov (United States)

Nomura, Yoshiaki; Takeuchi, Hiroaki; Okamoto, Masaaki; Sogabe, Kaoru; Okada, Ayako; Hanada, Nobuhiro

2017-06-01

Prevotella intermedia/nigrescens is one of the well-known pathogens causing periodontal diseases, and the red florescence excited by the visible blue light caused by the protoporphyrin IX in the bacterial cells could be useful for the chair-side detection. The aim of this study was to evaluated levels of periodontal pathogen, especially P. intermedia in clinical samples of red fluorescent dental plaque. Thirty two supra gingival plaque samples from six individuals were measured its fluorescence at 640nm wavelength excited by 409nm. Periodontopathic bacteria were counted by the Invader PLUS PCR assay. Co-relations the fluorescence intensity and bacterial counts were analyzed by Person's correlation coefficient and simple and multiple regression analysis. Positive and negative predictive values of the fluorescence intensities for with or without P. intermedia in supragingival plaque was calculated. When relative fluorescence unit (RFU) were logarithmic transformed, statistically significant linear relations between RFU and bacterial counts were obtained for P. intermedia, Porphyromonas gingivalis and Tannerella forsythia. By the multiple regression analysis, only P. intermedia had statistically significant co-relation with fluorescence intensities. All of the fluorescent dental plaque contained P. intermedia m. In contrast, 28% of non-fluorescent plaques contained P. intermedia. To check the fluorescence dental plaque in the oral cavity could be the simple chair-side screening of the mature dental plaque before examining the periodontal pathogens especially P. intermedia by the PCR method. Copyright © 2017 Elsevier B.V. All rights reserved.

Intense particle beam and multiple applications

International Nuclear Information System (INIS)

Ueda, M.; Machida, M.

1988-01-01

The Multiple Application Intense Particle Beam project is an experiment in which an injector of high energy neutral or ionized particles will be used to diagnose high density and high temperature plasmas. The acceleration of the particles will be carried out feeding a diode with a high voltage pulse produced by a Marx generator. Other apllications of intense particle beam generated by this injector that could be explored in the future include: heating and stabilization of compact toroids, treatment of metallic surfaces and ion implantation. (author) [pt

Analysis of Cell Movement by Simultaneous Quantification of Local Membrane Displacement and Fluorescent Intensities Using Quimp2

NARCIS (Netherlands)

Bosgraaf, Leonard; van Haastert, Peter J. M.; Bretschneider, Till

The use of fluorescent markers in living cells has increased dramatically in the recent years. The quantitative analysis of the images requires specific analysis software. Previously, the program Quimp was launched for quantitating fluorescent intensities at the membrane or the cortex of the cell.

Numerical evaluation of droplet sizing based on the ratio of fluorescent and scattered light intensities (LIF/Mie technique)

International Nuclear Information System (INIS)

Charalampous, Georgios; Hardalupas, Yannis

2011-01-01

The dependence of fluorescent and scattered light intensities from spherical droplets on droplet diameter was evaluated using Mie theory. The emphasis is on the evaluation of droplet sizing, based on the ratio of laser-induced fluorescence and scattered light intensities (LIF/Mie technique). A parametric study is presented, which includes the effects of scattering angle, the real part of the refractive index and the dye concentration in the liquid (determining the imaginary part of the refractive index). The assumption that the fluorescent and scattered light intensities are proportional to the volume and surface area of the droplets for accurate sizing measurements is not generally valid. More accurate sizing measurements can be performed with minimal dye concentration in the liquid and by collecting light at a scattering angle of 60 deg. rather than the commonly used angle of 90 deg. Unfavorable to the sizing accuracy are oscillations of the scattered light intensity with droplet diameter that are profound at the sidescatter direction (90 deg.) and for droplets with refractive indices around 1.4.

Segmentation-based retrospective shading correction in fluorescence microscopy E. coli images for quantitative analysis

Science.gov (United States)

Mai, Fei; Chang, Chunqi; Liu, Wenqing; Xu, Weichao; Hung, Yeung S.

2009-10-01

Due to the inherent imperfections in the imaging process, fluorescence microscopy images often suffer from spurious intensity variations, which is usually referred to as intensity inhomogeneity, intensity non uniformity, shading or bias field. In this paper, a retrospective shading correction method for fluorescence microscopy Escherichia coli (E. Coli) images is proposed based on segmentation result. Segmentation and shading correction are coupled together, so we iteratively correct the shading effects based on segmentation result and refine the segmentation by segmenting the image after shading correction. A fluorescence microscopy E. Coli image can be segmented (based on its intensity value) into two classes: the background and the cells, where the intensity variation within each class is close to zero if there is no shading. Therefore, we make use of this characteristics to correct the shading in each iteration. Shading is mathematically modeled as a multiplicative component and an additive noise component. The additive component is removed by a denoising process, and the multiplicative component is estimated using a fast algorithm to minimize the intra-class intensity variation. We tested our method on synthetic images and real fluorescence E.coli images. It works well not only for visual inspection, but also for numerical evaluation. Our proposed method should be useful for further quantitative analysis especially for protein expression value comparison.

Intensity ratio to improve black hole assessment in multiple sclerosis.

Science.gov (United States)

Adusumilli, Gautam; Trinkaus, Kathryn; Sun, Peng; Lancia, Samantha; Viox, Jeffrey D; Wen, Jie; Naismith, Robert T; Cross, Anne H

2018-01-01

Improved imaging methods are critical to assess neurodegeneration and remyelination in multiple sclerosis. Chronic hypointensities observed on T1-weighted brain MRI, "persistent black holes," reflect severe focal tissue damage. Present measures consist of determining persistent black holes numbers and volumes, but do not quantitate severity of individual lesions. Develop a method to differentiate black and gray holes and estimate the severity of individual multiple sclerosis lesions using standard magnetic resonance imaging. 38 multiple sclerosis patients contributed images. Intensities of lesions on T1-weighted scans were assessed relative to cerebrospinal fluid intensity using commercial software. Magnetization transfer imaging, diffusion tensor imaging and clinical testing were performed to assess associations with T1w intensity-based measures. Intensity-based assessments of T1w hypointensities were reproducible and achieved > 90% concordance with expert rater determinations of "black" and "gray" holes. Intensity ratio values correlated with magnetization transfer ratios (R = 0.473) and diffusion tensor imaging metrics (R values ranging from 0.283 to -0.531) that have been associated with demyelination and axon loss. Intensity ratio values incorporated into T1w hypointensity volumes correlated with clinical measures of cognition. This method of determining the degree of hypointensity within multiple sclerosis lesions can add information to conventional imaging. Copyright © 2017 Elsevier B.V. All rights reserved.

Amplification of the Signal Intensity of Fluorescence-Based Fiber-Optic Biosensors Using a Fabry-Perot Resonator Structure

Directory of Open Access Journals (Sweden)

Meng-Chang Hsieh

2015-02-01

Full Text Available Fluorescent biosensors have been widely used in biomedical applications. To amplify the intensity of fluorescence signals, this study developed a novel structure for an evanescent wave fiber-optic biosensor by using a Fabry-Perot resonator structure. An excitation light was coupled into the optical fiber through a laser-drilled hole on the proximal end of the resonator. After entering the resonator, the excitation light was reflected back and forth inside the resonator, thereby amplifying the intensity of the light in the fiber. Subsequently, the light was used to excite the fluorescent molecules in the reactive region of the sensor. The experimental results showed that the biosensor signal was amplified eight-fold when the resonator reflector was formed using a 92% reflective coating. Furthermore, in a simulation, the biosensor signal could be amplified 20-fold by using a 99% reflector.

Three-dimensional fluorescent microscopy via simultaneous illumination and detection at multiple planes.

Science.gov (United States)

Ma, Qian; Khademhosseinieh, Bahar; Huang, Eric; Qian, Haoliang; Bakowski, Malina A; Troemel, Emily R; Liu, Zhaowei

2016-08-16

The conventional optical microscope is an inherently two-dimensional (2D) imaging tool. The objective lens, eyepiece and image sensor are all designed to capture light emitted from a 2D 'object plane'. Existing technologies, such as confocal or light sheet fluorescence microscopy have to utilize mechanical scanning, a time-multiplexing process, to capture a 3D image. In this paper, we present a 3D optical microscopy method based upon simultaneously illuminating and detecting multiple focal planes. This is implemented by adding two diffractive optical elements to modify the illumination and detection optics. We demonstrate that the image quality of this technique is comparable to conventional light sheet fluorescent microscopy with the advantage of the simultaneous imaging of multiple axial planes and reduced number of scans required to image the whole sample volume.

Fluorescence of the gamma, epsilon, and delta systems of nitric oxide - Polarization and use of calculated intensities for spectrometer calibration.

Science.gov (United States)

Poland, H. M.; Broida, H. P.

1971-01-01

Results of a study in which fluorescence of the gamma system of nitric oxide was obtained by excitation from both the 2144 A line of ionized cadmium and a continuum source. Individual rotational lines of the 2144 A excited fluorescence spectrum were found to be partially polarized and to have polarizations of differ ing sign. Measured relative vibrational band intensities from line and continuum excitation were compared to calculated Franck-Condon factors. Those Franck-Condon factors based on a single potential for the two spin states of the X super pi state agreed better with measured values than those based on separate potentials for the two spin states. Calculated intensities of the v prime = 3 progression were used to calibrate the instrument response in the wavelength region from 2000 to 2500 A and were checked with measured intensities of the v prime = 0.1, and 2 progressions. Fluorescence of the epsilon and delta bands obtained with continuum lamp excitation also were compared to calculated intensities.

Fluorescent optical position sensor

Science.gov (United States)

Weiss, Jonathan D.

2005-11-15

A fluorescent optical position sensor and method of operation. A small excitation source side-pumps a localized region of fluorescence at an unknown position along a fluorescent waveguide. As the fluorescent light travels down the waveguide, the intensity of fluorescent light decreases due to absorption. By measuring with one (or two) photodetectors the attenuated intensity of fluorescent light emitted from one (or both) ends of the waveguide, the position of the excitation source relative to the waveguide can be determined by comparing the measured light intensity to a calibrated response curve or mathematical model. Alternatively, excitation light can be pumped into an end of the waveguide, which generates an exponentially-decaying continuous source of fluorescent light along the length of the waveguide. The position of a photodetector oriented to view the side of the waveguide can be uniquely determined by measuring the intensity of the fluorescent light emitted radially at that location.

Comparative study of the fluorescence intensity of dental composites and human teeth submitted to artificial aging.

Science.gov (United States)

Jablonski, Tatiana; Takahashi, Marcos Kenzo; Brum, Rafeal Torres; Rached, Rodrigo Nunes; Souza, Evelise M

2014-01-01

The aim of this study was to evaluate quantitatively the fluorescence of resin composites and human teeth, and to determine the stability of fluorescence after aging. Ten specimens were built using a 1 mm thick increment of dentin composite overlapped by a 0.5 mm thick increment of enamel composite. Ten sound human molars were sectioned and silicon carbide-polished to obtain enamel and dentin slabs 1.5 mm in thickness. Fluorescence measurements were carried out by a fluorescence spectrophotometer before and after thermocycling (2000 cycles, 5°C and 55°C). One-way analysis of variance (ANOVA) with repeated measures and Tukey's test were performed at a significance level of 5%. Most of the tested composites showed significant differences in fluorescence both before and after aging (P 0.05), and was the only composite that showed comparable results of fluorescence to the tooth structure before and after thermocycling. With the exception of Filtek Supreme, there were significant reductions in fluorescence intensity for all the tested composites (P < 0.05).

Human cytomegaloviruses expressing yellow fluorescent fusion proteins--characterization and use in antiviral screening.

Directory of Open Access Journals (Sweden)

Sarah Straschewski

Full Text Available Recombinant viruses labelled with fluorescent proteins are useful tools in molecular virology with multiple applications (e.g., studies on intracellular trafficking, protein localization, or gene activity. We generated by homologous recombination three recombinant cytomegaloviruses carrying the enhanced yellow fluorescent protein (EYFP fused with the viral proteins IE-2, ppUL32 (pp150, and ppUL83 (pp65. In growth kinetics, the three viruses behaved all like wild type, even at low multiplicity of infection (MOI. The expression of all three fusion proteins was detected, and their respective localizations were the same as for the unmodified proteins in wild-type virus-infected cells. We established the in vivo measurement of fluorescence intensity and used the recombinant viruses to measure inhibition of viral replication by neutralizing antibodies or antiviral substances. The use of these viruses in a pilot screen based on fluorescence intensity and high-content analysis identified cellular kinase inhibitors that block viral replication. In summary, these viruses with individually EYFP-tagged proteins will be useful to study antiviral substances and the dynamics of viral infection in cell culture.

Statistical model of intensity noise in con focal fluorescence microscopy images

International Nuclear Information System (INIS)

Montereali, R.M.; Almaviva, S.; Franzini, I.; Somma, F.

2008-01-01

The visible photoluminescence of aggregate F2 and F3+ color centers in Lithium Fluoride (LiF) thin layers, grown by thermal evaporation on various substrates (either crystalline or not) with different thicknesses, can be efficiently observed by using an optical con focal fluorescence microscope and a laser pump with emission wavelength tuned at about 450 nm. Starting from con focal fluorescence images of uniformly colored LiF samples, an automatic routine for the estimation of photoluminescence intensity noise has been developed at the Solid State Laser Laboratory and Spectroscopy of the ENEA Research Center in Frascati. We reported experimental results about application of that routine to the photoluminescence of LiF thin films, uniformly irradiated with an X-ray tube with energy spectrum centered on the Cu K? emission line (8,03 keV), at the CNR-IFN in Rome, that allow to identify a suitable statistical model for his description [it

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

Science.gov (United States)

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

2017-02-01

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

Handheld Fluorescence Microscopy based Flow Analyzer.

Science.gov (United States)

Saxena, Manish; Jayakumar, Nitin; Gorthi, Sai Siva

2016-03-01

Fluorescence microscopy has the intrinsic advantages of favourable contrast characteristics and high degree of specificity. Consequently, it has been a mainstay in modern biological inquiry and clinical diagnostics. Despite its reliable nature, fluorescence based clinical microscopy and diagnostics is a manual, labour intensive and time consuming procedure. The article outlines a cost-effective, high throughput alternative to conventional fluorescence imaging techniques. With system level integration of custom-designed microfluidics and optics, we demonstrate fluorescence microscopy based imaging flow analyzer. Using this system we have imaged more than 2900 FITC labeled fluorescent beads per minute. This demonstrates high-throughput characteristics of our flow analyzer in comparison to conventional fluorescence microscopy. The issue of motion blur at high flow rates limits the achievable throughput in image based flow analyzers. Here we address the issue by computationally deblurring the images and show that this restores the morphological features otherwise affected by motion blur. By further optimizing concentration of the sample solution and flow speeds, along with imaging multiple channels simultaneously, the system is capable of providing throughput of about 480 beads per second.

Comparative study of the fatty acid binding process of a new FABP from Cherax quadricarinatus by fluorescence intensity, lifetime and anisotropy.

Directory of Open Access Journals (Sweden)

Jiayao Li

Full Text Available Fatty acid-binding proteins (FABPs are small cytosolic proteins, largely distributed in invertebrates and vertebrates, which accomplish uptake and intracellular transport of hydrophobic ligands such as fatty acids. Although long chain fatty acids play multiple crucial roles in cellular functions (structural, energy metabolism, regulation of gene expression, the precise functions of FABPs, especially those of invertebrate species, remain elusive. Here, we have identified and characterized a novel FABP family member, Cq-FABP, from the hepatopancreas of red claw crayfish Cherax quadricarinatus. We report the characterization of fatty acid-binding affinity of Cq-FABP by four different competitive fluorescence-based assays. In the two first approaches, the fluorescent probe 8-Anilino-1-naphthalenesulfonate (ANS, a binder of internal cavities of protein, was used either by directly monitoring its fluorescence emission or by monitoring the fluorescence resonance energy transfer occurring between the single tryptophan residue of Cq-FABP and ANS. The third and the fourth approaches were based on the measurement of the fluorescence emission intensity of the naturally fluorescent cis-parinaric acid probe or the steady-state fluorescence anisotropy measurements of a fluorescently labeled fatty acid (BODIPY-C16, respectively. The four methodologies displayed consistent equilibrium constants for a given fatty acid but were not equivalent in terms of analysis. Indeed, the two first methods were complicated by the existence of non specific binding modes of ANS while BODIPY-C16 and cis-parinaric acid specifically targeted the fatty acid binding site. We found a relationship between the affinity and the length of the carbon chain, with the highest affinity obtained for the shortest fatty acid, suggesting that steric effects primarily influence the interaction of fatty acids in the binding cavity of Cq-FABP. Moreover, our results show that the binding affinities

Accurate thermometry based on the red and green fluorescence intensity ratio in NaYF4: Yb, Er nanocrystals for bioapplication.

Science.gov (United States)

Liu, Lixin; Qin, Feng; Lv, Tianquan; Zhang, Zhiguo; Cao, Wenwu

2016-10-15

A biological temperature measurement method based on the fluorescence intensity ratio (FIR) was developed to reduce uncertainty. The upconversion luminescence of NaYF4:Yb, Er nanocrystals was studied as a function of temperature around the physiologically relevant range of 300-330 K. We found that the green-green FIR Fe and red-green FIR (I660/I540) varied linearly as temperature increased. The thermometric uncertainties using the two FIRs were discussed and were determined to be almost constant at 0.6 and 0.09 K for green-green and red-green, respectively. The lower thermometric uncertainty comes from the intense signal-to-noise ratio of the measured FIRs owing to their comparable fluorescence intensities.

Biomolecule-to-fluorescent-color encoder: modulation of fluorescence emission via DNA structural changes

Science.gov (United States)

Nishimura, Takahiro; Ogura, Yusuke; Yamada, Kenji; Ohno, Yuko; Tanida, Jun

2014-01-01

A biomolecule-to-fluorescent-color (B/F) encoder for optical readout of biomolecular information is proposed. In the B/F encoder, a set of fluorescence wavelengths and their intensity levels are used for coding of a biomolecular signal. A hybridization chain reaction of hairpin DNAs labeled with fluorescent reporters was performed to generate the fluorescence color codes. The fluorescence is modulated via fluorescence resonance energy transfer, which is controlled by DNA structural changes. The results demonstrate that fluorescent color codes can be configured based on two wavelengths and five intensities using the B/F encoder, and the assigned codes can be retrieved via fluorescence measurements. PMID:25071950

A simple and sensitive method for L-cysteine detection based on the fluorescence intensity increment of quantum dots

International Nuclear Information System (INIS)

Huang Shan; Xiao Qi; Li Ran; Guan Hongliang; Liu Jing; Liu Xiaorong; He Zhike; Liu Yi

2009-01-01

In this contribution, a simple and sensitive method for L-cysteine detection was established based on the increment of the fluorescence intensity of mercaptoacetic acid-capped CdSe/ZnS quantum dots (QDs) in aqueous solution. Meanwhile, the fluorescence characteristics and the optimal conditions were investigated in detail. Under the optimized conditions, the linear range of QDs fluorescence intensity versus the concentration of L-cysteine was 10-800 nmol L -1 , with a correlation coefficient (R) of 0.9969 and a limit of detection (3σ black) of 3.8 nmol L -1 . The relative standard deviation (R.S.D.) for 0.5 μmol L -1 L-cysteine was 1.1% (n = 5). There was no interference to coexisting foreign substances including common ions, carbohydrates, nucleotide acids and other 19 amino acids. The proposed method possessed the advantages of simplicity, rapidity and sensitivity. Synthetic amino acid samples, medicine sample together with human urine samples were analyzed by the methodology and the results were satisfying.

Effects of water stress and light intensity on chlorophyll fluorescence parameters and pigments of Aloe vera L.

Science.gov (United States)

Hazrati, Saeid; Tahmasebi-Sarvestani, Zeinolabedin; Modarres-Sanavy, Seyed Ali Mohammad; Mokhtassi-Bidgoli, Ali; Nicola, Silvana

2016-09-01

Aloe vera L. is one of the most important medicinal plants in the world. In order to determine the effects of light intensity and water deficit stress on chlorophyll (Chl) fluorescence and pigments of A. vera, a split-plot in time experiment was laid out in a randomized complete block design with four replications in a research greenhouse. The factorial combination of three light intensities (50, 75 and 100% of sunlight) and four irrigation regimes (irrigation after depleting 20, 40, 60 and 80% of soil water content) were considered as main factors. Sampling time was considered as sub factor. The first, second and third samplings were performed 90, 180 and 270 days after imposing the treatments, respectively. The results demonstrated that the highest light intensity and the severe water stress decreased maximum fluorescence (Fm), variable fluorescence (Fv)/Fm, quantum yield of PSII photochemistry (ФPSII), Chl and photochemical quenching (qP) but increased non-photochemical quenching (NPQ), minimum fluorescence (F0) and Anthocyanin (Anth). Additionally, the highest Fm, Fv/Fm, ФPSII and qP and the lowest NPQ and F0 were observed when 50% of sunlight was blocked and irrigation was done after 40% soil water depletion. Irradiance of full sunlight and water deficit stress let to the photoinhibition of photosynthesis, as indicated by a reduced quantum yield of PSII, ФPSII, and qP, as well as higher NPQ. Thus, chlorophyll florescence measurements provide valuable physiological data. Close to half of total solar radiation and irrigation after depleting 40% of soil water content were selected as the most efficient treatments. Copyright © 2016 Elsevier Masson SAS. All rights reserved.

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

Directory of Open Access Journals (Sweden)

Guitao Zhong

2017-06-01

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

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

Science.gov (United States)

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

2009-01-01

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

Stage-specific fluorescence intensity of GFP and mCherry during sporulation In Bacillus Subtilis

Directory of Open Access Journals (Sweden)

Bailey Kirra

2010-11-01

Full Text Available Abstract Background Fluorescent proteins are powerful molecular biology tools that have been used to study the subcellular dynamics of proteins within live cells for well over a decade. Two fluorescent proteins commonly used to enable dual protein labelling are GFP (green and mCherry (red. Sporulation in the Gram positive bacterium Bacillus subtilis has been studied for many years as a paradigm for understanding the molecular basis for differential gene expression. As sporulation initiates, cells undergo an asymmetric division leading to differential gene expression in the small prespore and large mother cell compartments. Use of two fluorescent protein reporters permits time resolved examination of differential gene expression either in the same compartments or between compartments. Due to the spectral properties of GFP and mCherry, they are considered an ideal combination for co-localisation and co-expression experiments. They can also be used in combination with fluorescent DNA stains such as DAPI to correlate protein localisation patterns with the developmental stage of sporulation which can be linked to well characterised changes in DNA staining patterns. Findings While observing the recruitment of the transcription machinery into the forespore of sporulating Bacillus subtilis, we noticed the occurrence of stage-specific fluorescence intensity differences between GFP and mCherry. During vegetative growth and the initial stages of sporulation, fluorescence from both GFP and mCherry fusions behaved similarly. During stage II-III of sporulation we found that mCherry fluorescence was considerably diminished, whilst GFP signals remained clearly visible. This fluorescence pattern reversed during the final stage of sporulation with strong mCherry and low GFP fluorescence. These trends were observed in reciprocal tagging experiments indicating a direct effect of sporulation on fluorescent protein fluorophores. Conclusions Great care should be taken

A Solar-Pumped Fluorescence Model for Line-By-Line Emission Intensities in the B-X, A-X, and X-X Band Systems of 12C14N

Science.gov (United States)

Paganini, L.; Mumma, M. J.

2016-01-01

We present a new quantitative model for detailed solar-pumped fluorescent emission of the main isotopologue of CN. The derived fluorescence efficiencies permit estimation and interpretation of ro-vibrational infrared line intensities of CN in exospheres exposed to solar (or stellar) radiation. Our g-factors are applicable to astronomical observations of CN extending from infrared to optical wavelengths, and we compare them with previous calculations in the literature. The new model enables extraction of rotational temperature, column abundance, and production rate from astronomical observations of CN in the inner coma of comets. Our model accounts for excitation and de-excitation of rotational levels in the ground vibrational state by collisions, solar excitation to the A(sup 2)Pi(sub I) and B(sup 2)Sum(sup +) electronically excited states followed by cascade to ro-vibrational levels of X(sup 2)Sum(sup +), and direct solar infrared pumping of ro-vibrational levels in the X(sup 2)Sum(sup +) state. The model uses advanced solar spectra acquired at high spectral resolution at the relevant infrared and optical wavelengths and considers the heliocentric radial velocity of the comet (the Swings effect) when assessing the exciting solar flux for a given transition. We present model predictions for the variation of fluorescence rates with rotational temperature and heliocentric radial velocity. Furthermore, we test our fluorescence model by comparing predicted and measured line-by-line intensities for X(sup 2)Sum(sup +) (1-0) in comet C/2014 Q2 (Lovejoy), thereby identifying multiple emission lines observed at IR wavelengths.

Polar plot representation of time-resolved fluorescence.

Science.gov (United States)

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

2014-01-01

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

The relative-intensity method of X-ray fluorescence analysis and its application to soils and rocks

International Nuclear Information System (INIS)

Childs, C.W.; Furkert, R.J.

1974-01-01

The relative-intensity X-ray fluorescence method of analysis of rock and soil samples has been investigated and compared with the net-intensity method. Strong, coherently scattered radiation originating from the X-ray tube is shown to be preferable to background radiation as an internal standard, and scattered radiation measured at one wavelength can usefully be applied in the determination of several elements. When the concentrations of an element in two soil samples of different composition (for example concretions and the soil adjacent to them) are compared, the ratio of the relative intensities may be different from the ratio of net intensities by a factor of about two. The concentrations of manganese in thirteen standard rock samples determined by the relative-intensity method are within or very close to the ranges of values reported previously

Fluorescence intensity dependence on the propagation plane inclination

International Nuclear Information System (INIS)

Fernandez, J.E.; Rubio, Marcelo; Sanchez, H.J.

1987-01-01

A Monte Carlo simulation of the primary and secondary X-ray fluorescent emission from an homogeneous and infinite thickness sample, irradiated under different inclination of the propagation plane, is carried out. An agreement with the predictions based on Sherman equations depending on the inclination angle α was found. The invariance of the primary fluorescence with respect to α and the decrease until evanescence of the secondary fluorescence for a α → π/2 are confirmed. A discussion about the physical basis of this dependence is carried out. Similar results are expected for tertiary fluorescence. (Author) [es

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

Directory of Open Access Journals (Sweden)

Hairong Yin

2017-06-01

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

Fluorescence Behavior and Dural Infiltration of Meningioma Analyzed by 5-Aminolevulinic Acid-Based Fluorescence: Operating Microscope Versus Mini-Spectrometer.

Science.gov (United States)

Knipps, Johannes; Beseoglu, Kerim; Kamp, Marcel; Fischer, Igor; Felsberg, Joerg; Neumann, Lisa M; Steiger, Hans-Jakob; Cornelius, Jan F

2017-12-01

To compare fluorescence intensity of tumor specimens, as measured by a fluorescence-guided surgery microscope and a spectrometer, to evaluate tumor infiltration of dura mater around meningiomas with help of these 2 different 5-aminolevulinic acid (5-ALA)-based fluorescence tools, and to correlate fluorescence intensity with histopathologic data. In a clinical series, meningiomas were resected by 5-ALA fluorescence-guided surgery. Fluorescence intensity was semiquantitatively rated by the surgeon at predefined points. Biopsies were harvested and fluorescence intensity measured by a spectrometer and histopathologically analyzed. Sampling was realized at the level of the dura in a centrifugal direction. A total of 104 biopsies (n = 13 tumors) were analyzed. Specificity and sensitivity of the microscope were 0.96 and 0.53 and of the spectrometer 0.95 and 0.93, respectively. Fluorescence intensity as measured by the spectrometer was correlated to histologically confirmed tumor burden. In a centrifugal direction, tumor burden and fluorescence intensity continuously decreased (along the dural tail). Below a threshold value of 639 arbitrary units no tumor was histologically detectable. At the level of the dura the spectrometer was highly sensitive for detection of meningioma cells. The surgical microscope showed false negative results and missed residual tumor cells in more than one half of the cases. The complementary use of both fluorescence tools may improve resection quality. Copyright © 2017 Elsevier Inc. All rights reserved.

Synthesis and Fluorescence Spectra of Triazolylcoumarin Fluorescent Dyes

Institute of Scientific and Technical Information of China (English)

PENG Xian-fu; LI Hong-qi

2009-01-01

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

X-ray fluorescence holography and multiple-energy x-ray holography: A critical comparison of atomic images

International Nuclear Information System (INIS)

Len, P.M.; Gog, T.; Fadley, C.S.; Materlik, G.

1997-01-01

We compare x-ray fluorescence holography (XFH) and multiple-energy x-ray holography (MEXH), two techniques that have recently been used to obtain experimental three-dimensional atomic images. For single-energy holograms, these methods are equivalent by virtue of the optical reciprocity theorem. However, XFH can only record holographic information at the characteristic fluorescence energies of the emitting species, while MEXH can record holographic information at any energy above the fluorescent edge of the emitter, thus enabling the suppression of real-twin overlaps and other aberrations and artifacts in atomic images. copyright 1997 The American Physical Society

Noninvasive imaging of multiple myeloma using near infrared fluorescent molecular probe

Science.gov (United States)

Hathi, Deep; Zhou, Haiying; Bollerman-Nowlis, Alex; Shokeen, Monica; Akers, Walter J.

2016-03-01

Multiple myeloma is a plasma cell malignancy characterized by monoclonal gammopathy and osteolytic bone lesions. Multiple myeloma is most commonly diagnosed in late disease stages, presenting with pathologic fracture. Early diagnosis and monitoring of disease status may improve quality of life and long-term survival for multiple myeloma patients from what is now a devastating and fatal disease. We have developed a near-infrared targeted fluorescent molecular probe with high affinity to the α4β1 integrin receptor (VLA-4)overexpressed by a majority of multiple myeloma cells as a non-radioactive analog to PET/CT tracer currently being developed for human diagnostics. A near-infrared dye that emits about 700 nm was conjugated to a high affinity peptidomimmetic. Binding affinity and specificity for multiple myeloma cells was investigated in vitro by tissue staining and flow cytometry. After demonstration of sensitivity and specificity, preclinical optical imaging studies were performed to evaluate tumor specificity in murine subcutaneous and metastatic multiple myeloma models. The VLA-4-targeted molecular probe showed high affinity for subcutaneous MM tumor xenografts. Importantly, tumor cells specific accumulation in the bone marrow of metastatic multiple myeloma correlated with GFP signal from transfected cells. Ex vivo flow cytometry of tumor tissue and bone marrow further corroborated in vivo imaging data, demonstrating the specificity of the novel agent and potential for quantitative imaging of multiple myeloma burden in these models.

Fluorescence intensity dependence on the propagation plane inclination

International Nuclear Information System (INIS)

Fernandez, J.E.; Rubio, Marcelo; Sanchez, H.J.

1987-01-01

A theoretical study of the emission from layers of primary and secondary fluorescent components for a sample of infinite thickness was made, finding out that this emission depends mainly on the α angle as a maximum emission selector of a certain layer, which means 'tuning' the fluorescent radiation that comes primarily from a certain depth. These results can be applied to the study of both selective emission by layers and to the selection of superficial fluorescence. The analytical results have been confirmed by a Monte Carlo simulation. (Author) [es

Self-interference fluorescence microscopy with three-phase detection for depth-resolved confocal epi-fluorescence imaging.

Science.gov (United States)

Braaf, Boy; de Boer, Johannes F

2017-03-20

Three-dimensional confocal fluorescence imaging of in vivo tissues is challenging due to sample motion and limited imaging speeds. In this paper a novel method is therefore presented for scanning confocal epi-fluorescence microscopy with instantaneous depth-sensing based on self-interference fluorescence microscopy (SIFM). A tabletop epi-fluorescence SIFM setup was constructed with an annular phase plate in the emission path to create a spectral self-interference signal that is phase-dependent on the axial position of a fluorescent sample. A Mach-Zehnder interferometer based on a 3 × 3 fiber-coupler was developed for a sensitive phase analysis of the SIFM signal with three photon-counter detectors instead of a spectrometer. The Mach-Zehnder interferometer created three intensity signals that alternately oscillated as a function of the SIFM spectral phase and therefore encoded directly for the axial sample position. Controlled axial translation of fluorescent microsphere layers showed a linear dependence of the SIFM spectral phase with sample depth over axial image ranges of 500 µm and 80 µm (3.9 × Rayleigh range) for 4 × and 10 × microscope objectives respectively. In addition, SIFM was in good agreement with optical coherence tomography depth measurements on a sample with indocyanine green dye filled capillaries placed at multiple depths. High-resolution SIFM imaging applications are demonstrated for fluorescence angiography on a dye-filled capillary blood vessel phantom and for autofluorescence imaging on an ex vivo fly eye.

Potential rainfall-intensity and pH-driven shifts in the apparent fluorescent composition of dissolved organic matter in rainwater.

Science.gov (United States)

Zhou, Yongqiang; Yao, Xiaolong; Zhang, Yibo; Shi, Kun; Zhang, Yunlin; Jeppesen, Erik; Gao, Guang; Zhu, Guangwei; Qin, Boqiang

2017-05-01

Perturbations of rainwater chromophoric dissolved organic matter (CDOM) fluorescence induced by changes in rainfall intensity and pH were investigated by field observations and laboratory pH titrations. Microbial humic-like fluorophores dominated the rainwater CDOM pool, followed by tryptophan-like and tyrosine-like substances. Increased rainfall intensity had notable dilution effects on all six fluorescent components (C1-C6) identified using parallel factor (PARAFAC) analysis, the effect being especially pronounced for the microbial humic-like C1, tryptophan-like C3, and tyrosine-like C5. The results also indicated that increasing pH from 7 to 9 led to decreased fluorescence intensity (F max ) of all the six components, while a pH increase from 5 to 7, resulted in increasing F max of terrestrial humic-like C2, tyrosine-like C5, and tryptophan-like C6 and decreasing microbial humic-like C1, tryptophan-like C3, and fulvic-like C4. Two-dimensional correlation spectroscopy (2D-COS) demonstrated that synchronous fluorescence responded first to pH modifications at fulvic-like wavelength (λ Ex /λ Em  = ∼316/416 nm), followed by tyrosine-like wavelength (λ Ex /λ Em  = ∼204/304 nm), tryptophan-like wavelength (λ Ex /λ Em  = ∼226/326 nm), microbial humic-like wavelength (∼295/395 nm), and finally terrestrial humic-like wavelength (∼360/460 nm). Our results suggest that a decrease in areas affected by acid rain in South China occurring at present may possibly result in apparent compositional changes of CDOM fluorescence. The decreased rainfall in South-West China and increased rainfall in North-West China during the past five decades may possibly accordingly result in increased and decreased F max of all the six components identified in South-West and North-West China, respectively. Copyright © 2017 Elsevier Ltd. All rights reserved.

Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO₄(2-) Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System.

Science.gov (United States)

Chen, Jing; Ye, Wangquan; Guo, Jinjia; Luo, Zhao; Li, Ying

2016-07-13

A newly developed integrated fluorescence-Raman spectral system (λex = 532 nm) for detecting Chlorophyll-a (chl-a), Chromophoric Dissolved Organic Matter (CDOM), carotenoids and SO₄(2-) in situ was used to successfully investigate the diurnal variability of all above. Simultaneously using the integration of fluorescence spectroscopy and Raman spectroscopy techniques provided comprehensive marine information due to the complementarity between the different excitation mechanisms and different selection rules. The investigation took place in offshore seawater of the Yellow Sea (36°05'40'' N, 120°31'32'' E) in October 2014. To detect chl-a, CDOM, carotenoids and SO₄(2-), the fluorescence-Raman spectral system was deployed. It was found that troughs of chl-a and CDOM fluorescence signal intensity were observed during high tides, while the signal intensity showed high values with larger fluctuations during ebb-tide. Chl-a and carotenoids were influenced by solar radiation within a day cycle by different detection techniques, as well as displaying similar and synchronous tendency. CDOM fluorescence cause interference to the measurement of SO₄(2-). To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO₄(2-) concentration on the following day. The results demonstrated that the fluorescence-Raman spectral system has great potential in detection of chl-a, carotenoids, CDOM and SO₄(2-) in the ocean.

Fluorescence lifetime imaging of skin cancer

Science.gov (United States)

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

2011-03-01

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

Estimation of cerium and lanthanum content in core material of high intensity carbon arc electrodes by x-ray fluorescence method

International Nuclear Information System (INIS)

Nagpal, K.C.; Bhavalkar, R.H.

1977-01-01

The X-ray fluorescence method has been used to determine the weight percentages of cerium and lanthanum in the core material of high intensity carbon arc electrodes from the calibration curves plotted between the weight percentages of these elements and the peak-intensity ratios of CeLsub(α1), and LaLsub(α1) peaks to the neighbouring peak SnLsub(α1) due to an internal standard element. (author)

Fluorescent multiplex cell flow systems and methods

KAUST Repository

Merzaban, Jasmeen; Abuelela, Ayman F.; Mohammad, Amal Jehad

2017-01-01

scanning system emits multiple electromagnetic wavelengths simultaneously it cause multiple fluorescent labels having different excitation wavelength maximums to fluoresce. The system can simultaneously capture real-time fluorescence images from at least

Development of ultrasound-assisted fluorescence imaging of indocyanine green.

Science.gov (United States)

Morikawa, Hiroyasu; Toyota, Shin; Wada, Kenji; Uchida-Kobayashi, Sawako; Kawada, Norifumi; Horinaka, Hiromichi

2017-01-01

Indocyanine green (ICG) accumulation in hepatocellular carcinoma means tumors can be located by fluorescence. However, because of light scattering, it is difficult to detect ICG fluorescence from outside the body. We propose a new fluorescence imaging method that detects changes in the intensity of ICG fluorescence by ultrasound-induced temperature changes. ICG fluorescence intensity decreases as the temperature rises. Therefore, it should theoretically be possible to detect tissue distribution of ICG using ultrasound to heat tissue, moving the point of ultrasound transmission, and monitoring changes in fluorescence intensity. A new probe was adapted for clinical application. It consisted of excitation light from a laser, fluorescence sensing through a light pipe, and heating by ultrasound. We applied the probe to bovine liver to image the accumulation of ICG. ICG emits fluorescence (820 nm) upon light irradiation (783 nm). With a rise in temperature, the fluorescence intensity of ICG decreased by 0.85 %/°C. The distribution of fluorescent ICG was detected using an ultrasonic warming method in a new integrated probe. Modulating fluorescence by changing the temperature using ultrasound can determine where ICG accumulates at a depth, highlighting its potential as a means to locate hepatocellular carcinoma.

An expression for the atomic fluorescence and thermal-emission intensity under conditions of near saturation and arbitrary self-absorption

NARCIS (Netherlands)

Omenetto, N.; Winefordner, J.D.; Alkemade, C.T.J.

An expression for the effect of self-absorption on the fluorescence and thermal emission intensities is derived by taking into account stimulated emission. A simple, idealized case is considered, consisting of a two level atomic system, in a flame, homogeneous with respect to temperature and

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

Energy Technology Data Exchange (ETDEWEB)

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

1995-03-14

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

A fluorescence switch based on a controllable photochromic naphthopyran group

International Nuclear Information System (INIS)

Chen Lizhen; Wang Guang; Zhao Xiancai

2011-01-01

A fluorescence switch based on photoisomerization of naphthopyran (NP) has been designed by employing 2-(pyridin-2-yl)-benzimidazole (BPI) and the naphthopyran containing two pyran rings (NP) as fluorescent dye and photochromic compound, respectively. The fluorescence switch of benzimidazole derivative can be modulated either by controlling the irradiation time of UV light or by adjusting the amount ratio of fluorescent benzimidazole derivative to photochromic naphthopyran in both solution and polymethyl methacrylate (PMMA) film. The experimental results indicated that the decrease of fluorescence intensity of benzimidazole derivative is attributed to the interaction of benzimidazole with naphthopyran. - Highlights: → Naphthopyran was first used to fabricate fluorescence switch with benzimidazole derivative. → Fluorescence intensity can be modulated by controlling the UV irradiation time. → Fluorescence intensity can be adjusted by changing the ratio of benzimidazole derivative to naphthopyran. → Decrease of fluorescence intensity is attributed to the interaction of benzimidazole derivative and naphthopyran.

Time course of the rise in UV fluorescence intensity following irradiation and experimental conditions for the determination of this parameter in vital Ehrlich-ascitic tumour cells

International Nuclear Information System (INIS)

Merkle, K.

1977-01-01

The use of the impulse cytophotometric method of measurement in the UV region of the spectrum has allowed to determine the change of primary fluorescence intensity on vital unstained cells without the influence of UV exposure or other factors affecting the measuring result in an uncontrollable way. The studies on the influence of storage time and -temperature have shown that a storage time of up to 25 min at 20 0 C has no effect on the fluorescence intensity of Ehrlich ascites cells. On day 11 after inoculation the fluorescence level is about 20% higher than on day 8, but is much better reproducible at the latter date. After 600 rads gamma irradiation applied on the 6th day, this parameter starts oscillating around normal values and is stabilized within 20 hrs after irradiation; these values remain constant in the next 40 hrs. Even after exposure to 100, 200 and 300 rads gamma radiation a constant UV fluorescence intensity is registered in this time interval. Measurements of this parameter in the phase of relative constancy allow a clear-cut interpretation of the results of further radiobiological investigations. (author)

Active Appearance Segmentation for Intensity Inhomogeneity in Light Sheet Fluorescence Microscopy

DEFF Research Database (Denmark)

Jensen, Casper Bo; Lyksborg, Mark; Hecksher-Sørensen, J.

2016-01-01

inhomogeneities which are often seen in Light Sheet Fluorescence Microscopy (LSFM) images. This robustness is achieved by modelling the appearance of an image as a regularized Normalized Gradient Field (rNGF). We perform two experiments to challenge the model. First it is tested using a repeated leave......Active Appearance Models (AAM) are used for annotating or segmenting shapes in biomedical images. Performance relies heavily on the image data used to train the AAM. In this paper we improve the generalization properties of the model by making it robust to slowly varying spatial intensity......-one-out approach on images with minimal imperfections where the left out images are corrupted by a simulated bias field and segmented using the AAM. Secondly we test the model on LSFM images with common acquisition problems. In both experiments the proposed approach outperforms the often used AAM implementation...

Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO42− Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System

Science.gov (United States)

Chen, Jing; Ye, Wangquan; Guo, Jinjia; Luo, Zhao; Li, Ying

2016-01-01

A newly developed integrated fluorescence-Raman spectral system (λex = 532 nm) for detecting Chlorophyll-a (chl-a), Chromophoric Dissolved Organic Matter (CDOM), carotenoids and SO42− in situ was used to successfully investigate the diurnal variability of all above. Simultaneously using the integration of fluorescence spectroscopy and Raman spectroscopy techniques provided comprehensive marine information due to the complementarity between the different excitation mechanisms and different selection rules. The investigation took place in offshore seawater of the Yellow Sea (36°05′40′′ N, 120°31′32′′ E) in October 2014. To detect chl-a, CDOM, carotenoids and SO42−, the fluorescence-Raman spectral system was deployed. It was found that troughs of chl-a and CDOM fluorescence signal intensity were observed during high tides, while the signal intensity showed high values with larger fluctuations during ebb-tide. Chl-a and carotenoids were influenced by solar radiation within a day cycle by different detection techniques, as well as displaying similar and synchronous tendency. CDOM fluorescence cause interference to the measurement of SO42−. To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO42− concentration on the following day. The results demonstrated that the fluorescence-Raman spectral system has great potential in detection of chl-a, carotenoids, CDOM and SO42− in the ocean. PMID:27420071

Diurnal Variability in Chlorophyll-a, Carotenoids, CDOM and SO42− Intensity of Offshore Seawater Detected by an Underwater Fluorescence-Raman Spectral System

Directory of Open Access Journals (Sweden)

Jing Chen

2016-07-01

Full Text Available A newly developed integrated fluorescence-Raman spectral system (λex = 532 nm for detecting Chlorophyll-a (chl-a, Chromophoric Dissolved Organic Matter (CDOM, carotenoids and SO42− in situ was used to successfully investigate the diurnal variability of all above. Simultaneously using the integration of fluorescence spectroscopy and Raman spectroscopy techniques provided comprehensive marine information due to the complementarity between the different excitation mechanisms and different selection rules. The investigation took place in offshore seawater of the Yellow Sea (36°05′40′′ N, 120°31′32′′ E in October 2014. To detect chl-a, CDOM, carotenoids and SO42−, the fluorescence-Raman spectral system was deployed. It was found that troughs of chl-a and CDOM fluorescence signal intensity were observed during high tides, while the signal intensity showed high values with larger fluctuations during ebb-tide. Chl-a and carotenoids were influenced by solar radiation within a day cycle by different detection techniques, as well as displaying similar and synchronous tendency. CDOM fluorescence cause interference to the measurement of SO42−. To avoid such interference, the backup Raman spectroscopy system with λex = 785 nm was employed to detect SO42− concentration on the following day. The results demonstrated that the fluorescence-Raman spectral system has great potential in detection of chl-a, carotenoids, CDOM and SO42− in the ocean.

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

Science.gov (United States)

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

2006-10-01

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

Total Internal Reflection Fluorescence Microscopy Imaging-Guided Confocal Single-Molecule Fluorescence Spectroscopy

OpenAIRE

Zheng, Desheng; Kaldaras, Leonora; Lu, H. Peter

2013-01-01

We have developed an integrated spectroscopy system combining total internal reflection fluorescence microscopy imaging with confocal single-molecule fluorescence spectroscopy for two-dimensional interfaces. This spectroscopy approach is capable of both multiple molecules simultaneously sampling and in situ confocal fluorescence dynamics analyses of individual molecules of interest. We have demonstrated the calibration with fluorescent microspheres, and carried out single-molecule spectroscop...

Fluorescence Imaging Reveals Surface Contamination

Science.gov (United States)

Schirato, Richard; Polichar, Raulf

1992-01-01

In technique to detect surface contamination, object inspected illuminated by ultraviolet light to make contaminants fluoresce; low-light-level video camera views fluorescence. Image-processing techniques quantify distribution of contaminants. If fluorescence of material expected to contaminate surface is not intense, tagged with low concentration of dye.

Multiple temperature effects on up-conversion fluorescences of Er3+-Y b3+-Mo6+ codoped TiO2 and high thermal sensitivity

Directory of Open Access Journals (Sweden)

B. S. Cao

2015-08-01

Full Text Available We report multiple temperature effects on green and red up-conversion emissions in Er3+-Y b3+-Mo6+ codoped TiO2 phosphors. With increasing temperature, the decrease of the red emission from 4F9/2→4I15/2, the increase of green emission from 2H11/2→4I15/2 and another unchanged green emission from 4S3/2→4I15/2 were simultaneously observed, which are explained by steady-state rate equations analysis. Due to different evolution with temperature of the two green emissions, higher thermal sensitivity of optical thermal sensor was obtained based on the transitions with the largest fluorescence intensity ratio. Two parameters, maximum theoretical sensitivity (Smax and optimum operating temperature (Tmax are given to describe thermal sensing properties of the produced sensors. The intensity ratio and energy difference ΔE of a pair of energy levels are two main factors for the sensitivity and accuracy of sensors, which should be referred to design sensors with optimized sensing properties.

Multimodal fluorescence imaging spectroscopy

NARCIS (Netherlands)

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

2014-01-01

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

Impact of high-intensity concurrent training on cardiovascular risk factors in persons with multiple sclerosis - pilot study.

Science.gov (United States)

Keytsman, Charly; Hansen, Dominique; Wens, Inez; O Eijnde, Bert

2017-10-27

High-intensity concurrent training positively affects cardiovascular risk factors. Because this was never investigated in multiple sclerosis, the present pilot study explored the impact of this training on cardiovascular risk factors in this population. Before and after 12 weeks of high-intense concurrent training (interval and strength training, 5 sessions per 2 weeks, n = 16) body composition, resting blood pressure and heart rate, 2-h oral glucose tolerance (insulin sensitivity, glycosylated hemoglobin, blood glucose and insulin concentrations), blood lipids (high- and low-density lipoprotein, total cholesterol, triglyceride levels) and C-reactive protein were analyzed. Twelve weeks of high-intense concurrent training significantly improved resting heart rate (-6%), 2-h blood glucose concentrations (-13%) and insulin sensitivity (-24%). Blood pressure, body composition, blood lipids and C-reactive protein did not seem to be affected. Under the conditions of this pilot study, 12 weeks of concurrent high-intense interval and strength training improved resting heart rate, 2-h glucose and insulin sensitivity in multiple sclerosis but did not affect blood C-reactive protein levels, blood pressure, body composition and blood lipid profiles. Further, larger and controlled research investigating the effects of high-intense concurrent training on cardiovascular risk factors in multiple sclerosis is warranted. Implications for rehabilitation High-intensity concurrent training improves cardiovascular fitness. This pilot study explores the impact of this training on cardiovascular risk factors in multiple sclerosis. Despite the lack of a control group, high-intense concurrent training does not seem to improve cardiovascular risk factors in multiple sclerosis.

Wireless implantable electronic platform for chronic fluorescent-based biosensors.

Science.gov (United States)

Valdastri, Pietro; Susilo, Ekawahyu; Förster, Thilo; Strohhöfer, Christof; Menciassi, Arianna; Dario, Paolo

2011-06-01

The development of a long-term wireless implantable biosensor based on fluorescence intensity measurement poses a number of technical challenges, ranging from biocompatibility to sensor stability over time. One of these challenges is the design of a power efficient and miniaturized electronics, enabling the biosensor to move from bench testing to long term validation, up to its final application in human beings. In this spirit, we present a wireless programmable electronic platform for implantable chronic monitoring of fluorescent-based autonomous biosensors. This system is able to achieve extremely low power operation with bidirectional telemetry, based on the IEEE802.15.4-2003 protocol, thus enabling over three-year battery lifetime and wireless networking of multiple sensors. During the performance of single fluorescent-based sensor measurements, the circuit drives a laser diode, for sensor excitation, and acquires the amplified signals from four different photodetectors. In vitro functionality was preliminarily tested for both glucose and calcium monitoring, simply by changing the analyte-binding protein of the biosensor. Electronics performance was assessed in terms of timing, power consumption, tissue exposure to electromagnetic fields, and in vivo wireless connectivity. The final goal of the presented platform is to be integrated in a complete system for blood glucose level monitoring that may be implanted for at least one year under the skin of diabetic patients. Results reported in this paper may be applied to a wide variety of biosensors based on fluorescence intensity measurement.

Single Molecule Spectroscopy of Fluorescent Proteins

NARCIS (Netherlands)

Blum, Christian; Subramaniam, Vinod

2009-01-01

The discovery and use of fluorescent proteins has revolutionized cellular biology. Despite the widespread use of visible fluorescent proteins as reporters and sensors in cellular environments the versatile photophysics of fluorescent proteins is still subject to intense research. Understanding the

Fission products determination in high activity waste solution by wavelength dispersive X-ray fluorescence spectral interference correction by intensity ratio

International Nuclear Information System (INIS)

Sato, I.M.

1988-01-01

Fission products Se, Rb, Y, Zr, Mo, Ru, Rh, Pd, Te, Cd, Cs, Ba, La, Ce, Pr, Nd, Sm, Eu and Gd were determined in simulated high activity radioactive waste solution by wavelength dispersive X-ray fluorescence spectrometry without chemical separation. Thin layer technique was employed for the sample preparation. For the L spectral lines, the absorption effect was verified by Rasberry-Heinrich, Lucas Tooth-Pyne and Lachance-Trail relations. This effect was quantified and corrected accordingly. The spectral interferences of Kα and/or Lα lines of Y, Zr, Mo, La, Ce, Pr, Nd, Sm, Eu and Gd elements were eliminated by the intensity ratio method. The overlapping of up to three analytical lines was corrected by applying this method. The concentration influence of the interfering element on the intensity ratio values as well the efficiency of the correction method were investigated in order to assure that no systematic or residual error, resulting from the correction, affect the actual fluorescent intensity determination. The results is compared with the data obtained from measurements of free lines of spectral interference and also with those obtained by the linear equation system. Fission products determination presented a precision in the range of 0.1 to 5.0% and an accuracy of up to ± 7.0% the results are compared with those obtained by neutron activation analysis and inductively coupled plasma - atomic emission spectrometry. Leaching data, when radioactive waste is incorporated in cement matrix, were attempted by X-ray fluorescence technique. For two years leaching period, leaching rate and diffusion coefficient data of cesium were determined. The results obtained agree with those obtained by γ-spectromety. (author) [pt

Fluorescent sensors based on bacterial fusion proteins

International Nuclear Information System (INIS)

Mateu, Batirtze Prats; Pum, Dietmar; Sleytr, Uwe B; Toca-Herrera, José L; Kainz, Birgit

2014-01-01

Fluorescence proteins are widely used as markers for biomedical and technological purposes. Therefore, the aim of this project was to create a fluorescent sensor, based in the green and cyan fluorescent protein, using bacterial S-layers proteins as scaffold for the fluorescent tag. We report the cloning, expression and purification of three S-layer fluorescent proteins: SgsE-EGFP, SgsE-ECFP and SgsE-13aa-ECFP, this last containing a 13-amino acid rigid linker. The pH dependence of the fluorescence intensity of the S-layer fusion proteins, monitored by fluorescence spectroscopy, showed that the ECFP tag was more stable than EGFP. Furthermore, the fluorescent fusion proteins were reassembled on silica particles modified with cationic and anionic polyelectrolytes. Zeta potential measurements confirmed the particle coatings and indicated their colloidal stability. Flow cytometry and fluorescence microscopy showed that the fluorescence of the fusion proteins was pH dependent and sensitive to the underlying polyelectrolyte coating. This might suggest that the fluorescent tag is not completely exposed to the bulk media as an independent moiety. Finally, it was found out that viscosity enhanced the fluorescence intensity of the three fluorescent S-layer proteins. (paper)

Kα resonance fluorescence in Al, Ti, Cu and potential applications for X-ray sources

Science.gov (United States)

Nahar, Sultana N.; Pradhan, Anil K.

2015-04-01

The Kα resonance fluorescence (RFL) effect via photoabsorptions of inner shell electrons as the element goes through multiple ionization states is studied. We demonstrate that the resonances observed recently in Kα (1s-2p) fluorescence in aluminum plasmas by using a high-intensity X-ray free-electron laser [1] are basically K-shell resonances in hollow atoms going through multiple ionization states at resonant energies as predicted earlier for gold and iron ions [2]. These resonances are formed below the K-shell ionization edge and shift toward higher energies with ionization states, as observed. Fluorescence emission intensities depend on transition probabilities for each ionization stage of the given element for all possible Kα (1 s → 2 p) transition arrays. The present calculations for resonant photoabsorptions of Kα photons in Al have reproduced experimentally observed features. Resonant cross sections and absorption coefficients are presented for possible observation of Kα RFL in the resonant energy ranges of 4.5-5.0 keV for Ti ions and 8.0-8.7 keV for Cu ions respectively. We suggest that theoretically the Kα RFL process may be driven to enhance the Auger cycle by a twin-beam monochromatic X-ray source, tuned to the K-edge and Kα energies, with potential applications such as the development of narrow-band biomedical X-ray devices.

Synthesis of novel fluorescent probe Tb(III)-7-carboxymethoxy-4-methylcoumarin complex for sensing of DNA

Energy Technology Data Exchange (ETDEWEB)

Hussein, Belal H.M., E-mail: belalhussein102@yahoo.com [Department of Chemistry, Faculty of Science, Suez Canal University, Ismailia (Egypt); Azab, Hassan A. [Department of Chemistry, Faculty of Science, Suez Canal University, Ismailia (Egypt); Fathalla, Walid [Department of Mathematical and Physical Sciences, Faculty of Engineering, Port-Said University, Port-Said (Egypt); Ali, Sherin A.M. [Department of Mathematical and Physical Sciences, Faculty of Engineering, Suez Canal University, Ismailia (Egypt)

2013-02-15

New fluorescent probe Tb(III) (7-carboxymethoxy-4-methylcoumarin)2(SCN) (C2H5OH)(H2O) was synthesized and characterized by spectroscopy and thermal analysis. The absorption and fluorescence spectra of 7-carboxymethoxy-4-methylcoumarin (CMMC) and Tb(III)-CMMC complex have been measured in different solvents. The interactions of Tb(III)-CMMC complex with calf thymus nucleic acid (CT-DNA) have been investigated using steady state fluorescence measurements. The changes in the fluorescence intensity have been used for the quantitative determination of DNA with LOD of 3.45 ng in methanol-water (9:1, v/v). The association constants of DNA with Tb(III)-CMMC complex was found to be 2.62 Multiplication-Sign 1010 M{sup -1}. - Highlights: Black-Right-Pointing-Pointer New fluorescent probe Terbium (III)-7-carboxy methoxy-4-methylcoumarin complex has been synthesized and characterized. Black-Right-Pointing-Pointer FTIR spectrum of Tb(III)-complex shows a characteristic band for thiocyanate group. Black-Right-Pointing-Pointer DNA interaction with Terbium (III)-7-carboxy methoxy-4-methylcoumarin has been studied by fluorescence techniques. Black-Right-Pointing-Pointer The change in the fluorescence intensity has been used for the quantitative determination of DNA. Black-Right-Pointing-Pointer The result was better than most of the well-known methods including the ethidium bromide method.

Study of influencing factors to chromophoric dissolved organic matter absorption properties from fluorescence features in Taihu lake in autumn

Directory of Open Access Journals (Sweden)

Chuang-Chun Huang

2013-04-01

Full Text Available In order to identify the components of chromophoric dissolved organic matter (CDOM, confirm the influence of components to the absorption coefficient of CDOM (aCDOM, and estimate aCDOM from fluorescence spectra, fluorescence and optical measurements of CDOM were carried out in November 2008. The results indicate that, the primary component of CDOM is humic-like. The secondary component is tryptophan-like, which is the product of phytoplankton and aquatic debris rather than the wastewater treatment drainaged from city. In this study, six fluorophores with multiple excitation-emission matrices (EEMs peaks (A, B, C, N, M, T were identified according to the parallel factor analysis (PARAFAC. The average contribution of each component to the CDOM is 19.93, 18.82, 16.88, 16.39, 12.26, and 15.72%, respectively. Red Shifted phenomenon will happen with the increase of fluorescence intensity for ultraviolet and terrestrially humic-like. Conversely, marine humic-like will appear Reverse Red Shifted with the increase of fluorescence intensity. The primary contributor to the shoulder value of CDOM’s absorption coefficient at 275 nm is phytoplankton productivity, followed by marine humic-like. The main contributors to the shoulder shape are UV humic-like and phytoplankton productivity, followed by marine humic-like and tryptophan-like. A strong correlation between CDOM absorption and fluorescence intensity at emission wavelength of 424 nm and excitation wavelength ranging from 280 to 360 nm was found. The absorption coefficient can be retrieved successfully from the same excitation wavelength’s fluorescence intensity by an exponential model.

Reproducibility and Reliability of Repeated Quantitative Fluorescence Angiography

DEFF Research Database (Denmark)

Nerup, Nikolaj; Knudsen, Kristine Bach Korsholm; Ambrus, Rikard

2017-01-01

INTRODUCTION: When using fluorescence angiography (FA) in perioperative perfusion assessment, repeated measures with re-injections of fluorescent dye (ICG) may be required. However, repeated injections may cause saturation of dye in the tissue, exceeding the limit of fluorescence intensity...... that the camera can detect. As the emission of fluorescence is dependent of the excitatory light intensity, reduction of this may solve the problem. The aim of the present study was to investigate the reproducibility and reliability of repeated quantitative FA during a reduction of excitatory light....

Dual-detection confocal fluorescence microscopy: fluorescence axial imaging without axial scanning.

Science.gov (United States)

Lee, Dong-Ryoung; Kim, Young-Duk; Gweon, Dae-Gab; Yoo, Hongki

2013-07-29

We propose a new method for high-speed, three-dimensional (3-D) fluorescence imaging, which we refer to as dual-detection confocal fluorescence microscopy (DDCFM). In contrast to conventional beam-scanning confocal fluorescence microscopy, where the focal spot must be scanned either optically or mechanically over a sample volume to reconstruct a 3-D image, DDCFM can obtain the depth of a fluorescent emitter without depth scanning. DDCFM comprises two photodetectors, each with a pinhole of different size, in the confocal detection system. Axial information on fluorescent emitters can be measured by the axial response curve through the ratio of intensity signals. DDCFM can rapidly acquire a 3-D fluorescent image from a single two-dimensional scan with less phototoxicity and photobleaching than confocal fluorescence microscopy because no mechanical depth scans are needed. We demonstrated the feasibility of the proposed method by phantom studies.

Boundary segmentation for fluorescence microscopy using steerable filters

Science.gov (United States)

Ho, David Joon; Salama, Paul; Dunn, Kenneth W.; Delp, Edward J.

2017-02-01

Fluorescence microscopy is used to image multiple subcellular structures in living cells which are not readily observed using conventional optical microscopy. Moreover, two-photon microscopy is widely used to image structures deeper in tissue. Recent advancement in fluorescence microscopy has enabled the generation of large data sets of images at different depths, times, and spectral channels. Thus, automatic object segmentation is necessary since manual segmentation would be inefficient and biased. However, automatic segmentation is still a challenging problem as regions of interest may not have well defined boundaries as well as non-uniform pixel intensities. This paper describes a method for segmenting tubular structures in fluorescence microscopy images of rat kidney and liver samples using adaptive histogram equalization, foreground/background segmentation, steerable filters to capture directional tendencies, and connected-component analysis. The results from several data sets demonstrate that our method can segment tubular boundaries successfully. Moreover, our method has better performance when compared to other popular image segmentation methods when using ground truth data obtained via manual segmentation.

Impact of laser excitation intensity on deep UV fluorescence detection in microchip electrophoresis.

Science.gov (United States)

Schulze, Philipp; Ludwig, Martin; Belder, Detlev

2008-12-01

A high intensity 266 nm continuous wave (cw-) laser developed for material processing was utilised as an excitation source for sensitive native fluorescence detection of unlabelled compounds in MCE. This 120 mW laser was attached via an optical fibre into a commercial epifluorescence microscope. With this MCE set-up we evaluated the impact of laser power on the S/N of aromatic compounds as well as of proteins. Compared with a previous work which used a 4 mW pulsed laser for excitation, improved S/N for small aromatics and to a lesser extent for proteins could be attained. The LOD of the system was determined down to 24 ng/mL for serotonin (113 nM), 24 ng/mL for propranolol (81 nM), 80 ng/mL for tryptophan (392 nM) and 80 ng/mL for an aromatic diol (475 nM). Sensitive protein detection was obtained at concentrations of 5 microg/mL for lysocyme, trypsinogen and chymotrypsinogen (340, 208 and 195 nM, respectively). Finally, a comparison of the cw- with a pulsed 266 nm laser, operating at the same average power, showed a higher attainable sensitivity of the cw-laser. This can be attributed to fluorescence saturation and photobleaching effects of the pulsed laser at high pulse energies.

Fluorescent standards for photodynamic therapy

Science.gov (United States)

Belko, N.; Kavalenka, S.; Samtsov, M.

2016-08-01

Photodynamic therapy is an evolving technique for treatment of various oncological diseases. This method employs photosensitizers - species that lead to death of tumor cells after the photoactivation. For further development and novel applications of photodynamic therapy new photosensitizers are required. After synthesis of a new photosensitizer it is important to know its concentration in different biological tissues after its administration and distribution. The concentration is frequently measured by the extraction method, which has some disadvantages, e.g. it requires many biological test subjects that are euthanized during the measurement. We propose to measure the photosensitizer concentration in tissue by its fluorescence. For this purpose fluorescent standards were developed. The standards are robust and simple to produce; their fluorescence signal does not change with time. The fluorescence intensity of fluorescent standards seems to depend linearly on the dye concentration. A set of standards thus allow the calibration of a spectrometer. Finally, the photosensitizer concentration can be determined by the fluorescence intensity after comparing the corresponding spectrum with spectra of the set of fluorescent standards. A biological test subject is not euthanized during this kind of experiment. We hope this more humane technique can be used in future instead of the extraction method.

Measurement of protein-like fluorescence in river and waste water using a handheld spectrophotometer.

Science.gov (United States)

Baker, Andy; Ward, David; Lieten, Shakti H; Periera, Ryan; Simpson, Ellie C; Slater, Malcolm

2004-07-01

Protein-like fluorescence intensity in rivers increases with increasing anthropogenic DOM inputs from sewerage and farm wastes. Here, a portable luminescence spectrophotometer was used to investigate if this technology could be used to provide both field scientists with a rapid pollution monitoring tool and process control engineers with a portable waste water monitoring device, through the measurement of river and waste water tryptophan-like fluorescence from a range of rivers in NE England and from effluents from within two waste water treatment plants. The portable spectrophotometer determined that waste waters and sewerage effluents had the highest tryptophan-like fluorescence intensity, urban streams had an intermediate tryptophan-like fluorescence intensity, and the upstream river samples of good water quality the lowest tryptophan-like fluorescence intensity. Replicate samples demonstrated that fluorescence intensity is reproducible to +/- 20% for low fluorescence, 'clean' river water samples and +/- 5% for urban water and waste waters. Correlations between fluorescence measured by the portable spectrophotometer with a conventional bench machine were 0.91; (Spearman's rho, n = 143), demonstrating that the portable spectrophotometer does correlate with tryptophan-like fluorescence intensity measured using the bench spectrophotometer.

Parallel scan hyperspectral fluorescence imaging system and biomedical application for microarrays

International Nuclear Information System (INIS)

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

2011-01-01

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

Fluorescence Spectra Studies on the Interaction between Lanthanides and Calmodulin

Institute of Scientific and Technical Information of China (English)

无

1999-01-01

The conformation of Calmodulin(CaM) induced by lanthanides has been examined using fluorescence methods.With the addition of lanthanide (Ln3+), the intrinsic fluorescence intensity of CaM without calcium ions (Apo-CaM) first increases and then decreases.Ln3+ causes the decrease of intrinsic fluorescence intensity of calcium saturated CaM (Ca2+4-CaM) only at high concentrations.At low concentrations, Ln3+ results not only in the enhancement of fluorescence intensity of Apo-CaM, but also in a blue shift of the maximum emission wavelengh of dansyl labeled calmodulin(Apo-D-CaM).The molecular mechanism of the interaction between Ln3+ and CaM has been discussed in the light of the fluorescence spectra.

Simultaneous fluorescent detection of multiple metal ions based on the DNAzymes and graphene oxide.

Science.gov (United States)

Yun, Wen; Wu, Hong; Liu, Xingyan; Fu, Min; Jiang, Jiaolai; Du, Yunfeng; Yang, Lizhu; Huang, Yu

2017-09-15

A novel fluorescent detection strategy for simultaneous detection of Cu 2+ , Pb 2+ and Mg 2+ based on DNAzyme branched junction structure with three kinds of DNAzymes and graphene oxide (GO) was presented. Three fluorophores labeled DNA sequences consisted with enzyme-strand (E-DNA) and substrate strand (S-DNA) were annealed to form DNAzyme branched junction structure. In the presence of target metal ion, the DNAzyme was activated to cleave the fluorophore labeled S-DNA. The S-DNA fragments were released and adsorbed onto GO surface to quench the fluorescent signal. The detection limit was calculated to be 1 nM for Cu 2+ , 200 nM for Mg 2+ , and 0.3 nM for Pb 2+ , respectively. This strategy was successfully used for simultaneous detection of Cu 2+ , Mg 2+ and Pb 2+ in human serum. Moreover, it had potential application for simultaneous detection of multiple metal ions in environmental and biological samples. Copyright © 2017 Elsevier B.V. All rights reserved.

Fluorescent and high intensity discharge lamp use in chambers and greenhouses

Energy Technology Data Exchange (ETDEWEB)

Langhans, R.W. [Cornell Univ., Ithaca, NY (United States)

1994-12-31

Fluorescent and High Intensity Discharge lamps have opened up great opportunities for researchers to study plant growth under controlled environment conditions and for commercial growers to increase plant production during low/light periods. This report describes the advantages and disadvantages of using each lamp in growth chambers, growth rooms and greenhouses. Growth Chambers are small (3m x 4/m and smaller) walk-in or reach-in enclosures with programmable, accurate temperature, relative humidity (RH) and irradiance control over wide ranges. The intent of growth chambers was to replicate sunlight conditions and transfer research results directly to the greenhouse or outside. It was realized that sunlight and outside conditions could not be mimicked. Growth chambers are also used to study irradiance and spectral fluxes. Growth Rooms are usually large rooms (larger than 3m x 4m) with only lamp irradiance, but providing relatively limited ranges of environmental control (i.e., 10 to 30 C temperature, 50 to 90% RH and ambient to 1000 ppm CO{sub 2}), and commonly independent of outside conditions. Irradiance requirements for growth rooms are similar to those of growth chambers. Growth rooms are also used for growing a large number of plants in a uniform standard environment condition and in commercial horticulture for tissue culture, seed germination (plugs) and seedling growth. Greenhouses are designed to allow maximum sunlight penetration through the structure. Initially greenhouses were used to extend the growing season. Then as heating systems, and cooling systems improved, they were used year round. Low light during the winter months reduced plant growth, but with the advent of efficient lamps (HID and fluorescent) it became possible to increase growth to rates close to that in summer months. Supplementary lighting is used during low light periods of the year and anytime to ensure consistent total daily irradiance for research plants.

High intensity region segmentation in MR imaging of multiple sclerosis

International Nuclear Information System (INIS)

Rodrigo, F; Filipuzzi, M; Graffigna, J P; Isoardi, R; Noceti, M

2013-01-01

Numerous pathologies are often manifest in Magnetic Resonance Imaging (MRI) as hyperintense or bright regions as compared to normal tissue. It is of particular interest to develop an algorithm to detect, identify and define those Regions of Interest (ROI) when analyzing MRI studies, particularly for lesions of Multiple Sclerosis (MS). The objective of this study is to analyze those parameters which optimize segmentation of the areas of interest. To establish which areas should be considered as hyperintense regions, we developed a database (DB), with studies of patients diagnosed with MS. This disease causes axonal demyelination and it is expressed as bright regions in PD, T2 and FLAIR MRI sequences. Thus, with more than 4300 hyperintense regions validated by an expert physician, an algorithm was developed to detect such spots, approximating the results the expert obtained. Alongside these hyperintense lesion regions, it also detected bone regions with high intensity levels, similar to the intensity of the lesions, but with other features that allow a good differentiation.The algorithm will then detect ROIs with similar intensity levels and performs classification through data mining techniques

A fluorescence spectroscopy study of traditional Chinese medicine Angelica

Science.gov (United States)

Zhao, Hongyan; Song, Feng; Liu, Shujing; Chen, Guiyang; Wei, Chen; Liu, Yanling; Liu, Jiadong

2013-10-01

By measuring the fluorescence spectra of Chinese medicine (CM) Angelica water solutions with different concentrations from 0.025 to 2.5 mg/mL, results showed that the fluorescence intensity was proportional to the concentration. Through fluorescence spectra of Angelica solution under different pH values, results indicated coumarin compounds were the active ingredients of Angelica. We also observed fluorescence quenching of the Angelica solution in the presence of spherical silver nanoparticles with radius of 12 nm. Keeping a certain value for the volume of the silver nanoparticles, the fluorescence intensity at 402 nm was linearly proportional to the Angelica in the range of 1-3 mg/mL.

Saturated excitation of Fluorescence to quantify excitation enhancement in aperture antennas

KAUST Repository

Aouani, Heykel

2012-07-23

Fluorescence spectroscopy is widely used to probe the electromagnetic intensity amplification on optical antennas, yet measuring the excitation intensity amplification is a challenge, as the detected fluorescence signal is an intricate combination of excitation and emission. Here, we describe a novel approach to quantify the electromagnetic amplification in aperture antennas by taking advantage of the intrinsic non linear properties of the fluorescence process. Experimental measurements of the fundamental f and second harmonic 2f amplitudes of the fluorescence signal upon excitation modulation are used to quantify the electromagnetic intensity amplification with plasmonic aperture antennas. © 2012 Optical Society of America.

Saturated excitation of Fluorescence to quantify excitation enhancement in aperture antennas

KAUST Repository

Aouani, Heykel; Hostein, Richard; Mahboub, Oussama; Devaux, Eloï se; Rigneault, Hervé ; Ebbesen, Thomas W.; Wenger, Jé rô me

2012-01-01

Fluorescence spectroscopy is widely used to probe the electromagnetic intensity amplification on optical antennas, yet measuring the excitation intensity amplification is a challenge, as the detected fluorescence signal is an intricate combination of excitation and emission. Here, we describe a novel approach to quantify the electromagnetic amplification in aperture antennas by taking advantage of the intrinsic non linear properties of the fluorescence process. Experimental measurements of the fundamental f and second harmonic 2f amplitudes of the fluorescence signal upon excitation modulation are used to quantify the electromagnetic intensity amplification with plasmonic aperture antennas. © 2012 Optical Society of America.

Quantitative Brightness Analysis of Fluorescence Intensity Fluctuations in E. Coli.

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Kwang-Ho Hur

Full Text Available The brightness measured by fluorescence fluctuation spectroscopy specifies the average stoichiometry of a labeled protein in a sample. Here we extended brightness analysis, which has been mainly applied in eukaryotic cells, to prokaryotic cells with E. coli serving as a model system. The small size of the E. coli cell introduces unique challenges for applying brightness analysis that are addressed in this work. Photobleaching leads to a depletion of fluorophores and a reduction of the brightness of protein complexes. In addition, the E. coli cell and the point spread function of the instrument only partially overlap, which influences intensity fluctuations. To address these challenges we developed MSQ analysis, which is based on the mean Q-value of segmented photon count data, and combined it with the analysis of axial scans through the E. coli cell. The MSQ method recovers brightness, concentration, and diffusion time of soluble proteins in E. coli. We applied MSQ to measure the brightness of EGFP in E. coli and compared it to solution measurements. We further used MSQ analysis to determine the oligomeric state of nuclear transport factor 2 labeled with EGFP expressed in E. coli cells. The results obtained demonstrate the feasibility of quantifying the stoichiometry of proteins by brightness analysis in a prokaryotic cell.

Fluorescence detection system for microfluidic droplets

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

2018-05-01

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

Multiple high-intensity focused ultrasound probes for kidney-tissue ablation.

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Häcker, Axel; Chauhan, Sunita; Peters, Kristina; Hildenbrand, Ralf; Marlinghaus, Ernst; Alken, Peter; Michel, Maurice Stephan

2005-10-01

To investigate kidney-tissue ablation by high-intensity focused ultrasound (HIFU) using multiple and single probes. Ultrasound beams (1.75 MHz) produced by a piezoceramic element (focal distance 80 mm) were focused at the center of renal parenchyma. One of the three probes (mounted on a jig) could also be used for comparison with a single probe at comparable power ratings. Lesion dimensions were examined in perfused and unperfused ex vivo porcine kidneys at different power levels (40, 60, and 80 W) and treatment times (4, 6, and 8 seconds). At identical power levels, the lesions induced by multiple probes were larger than those induced by a single probe. Lesion size increased with increasing pulse duration and generator power. The sizes and shapes of the lesions were predictably repeatable in all samples. Lesions in perfused kidneys were smaller than those in unperfused kidneys. Ex vivo, kidney-tissue ablation by means of multiple HIFU probes offers significant advantages over single HIFU probes in respect of lesion size and formation. These advantages need to be confirmed by tests in vivo at higher energy levels.

Spectroscopic quantification of 5-hydroxymethylcytosine in genomic DNA using boric acid-functionalized nano-microsphere fluorescent probes.

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Chen, Hua-Yan; Wei, Jing-Ru; Pan, Jiong-Xiu; Zhang, Wei; Dang, Fu-Quan; Zhang, Zhi-Qi; Zhang, Jing

2017-05-15

5-hydroxymethylcytosine (5hmC) is the sixth base of DNA. It is involved in active DNA demethylation and can be a marker of diseases such as cancer. In this study, we developed a simple and sensitive 2-(4-boronophenyl)quinoline-4-carboxylic acid modified poly (glycidyl methacrylate (PBAQA-PGMA) fluorescent probe to detect the 5hmC content of genomic DNA based on T4 β-glucosyltransferase-catalyzed glucosylation of 5hmC. The fluorescence-enhanced intensity recorded from the DNA sample was proportional to its 5-hydroxymethylcytosine content and could be quantified by fluorescence spectrophotometry. The developed probe showed good detection sensitivity and selectivity and a good linear relationship between the fluorescence intensity and the concentration of 5 hmC within a 0-100nM range. Compared with other fluorescence detection methods, this method not only could determine trace amounts of 5 hmC from genomic DNA but also could eliminate the interference of fluorescent dyes and the need for purification. It also could avoid multiple labeling. Because the PBAQA-PGMA probe could enrich the content of glycosyl-5-hydroxymethyl-2-deoxycytidine from a complex ground substance, it will broaden the linear detection range and improve sensitivity. The limit of detection was calculated to be 0.167nM after enrichment. Furthermore, the method was successfully used to detect 5-hydroxymethylcytosine from mouse tissues. Copyright © 2016 Elsevier B.V. All rights reserved.

Fluorescent S-layer fusion proteins

International Nuclear Information System (INIS)

Kainz, B.

2010-01-01

This work describes the construction and characterisation of fluorescent S-layer fusion proteins used as building blocks for the fabrication of nanostructured monomolecular biocoatings on silica particles with defined fluorescence properties. The S-layer protein SgsE of Geobacillus stearothermophilus NRS 2004/3a was fused with the pH-dependant cyan, green and yellow variant of the green fluorescent protein (GFP) and the red fluorescent protein mRFP1. These fluorescent S-layer fusion proteins, acting as scaffold and optical sensing element simultaneously, were able to reassemble in solution and on silica particles forming 2D nanostructures with p2 lattice symmetry (a=11 ±0.5 nm, b=14 ±0.4 nm, g=80 ±1 o ). The pH-dependant fluorescence behaviour was studied with fluorimetry, confocal microscopy and flow cytometry. These fluorescent S-layer fusion proteins can be used as pH-sensor. 50% of the fluorescence intensity decreases at their calculated pKa values (pH6 - pH5). The fluorescence intensity of the GFP variants vanished completely between pH4 and pH3 whereas the chromophore of the red protein mRFP1 was only slightly affected in acidic conditions. At the isoelectric point of the S-layer coated silica particles (pH4.6 ±0.2) an increase in particle aggregation was detected by flow cytometry. The cyan and yellow fluorescent proteins were chosen to create a bi-fluorescent S-layer tandem fusion protein with the possibility for resonance energy transfer (FRET). A transfer efficiency of 20% and a molecular distance between the donor (ECFP) and acceptor (YFP) chromophores of around 6.2 nm could be shown. This bi-fluorescent ECFP-SgsE-YFP tandem fusion protein was able to reassemble on solid surfaces. The remarkable combination of fluorescence and self-assembly and the design of bi-functional S-layer tandem fusion protein matrices makes them to a promising tool in nanobiotechnology. (author) [de

Fluorescent property of 3-hydroxymethyl imidazo[1,2-a]pyridine and pyrimidine derivatives

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Velázquez-Olvera Stephania

2012-08-01

Full Text Available Abstract Background Imidazo[1,2-a]pyridines and pyrimidines are important organic fluorophores which have been investigated as biomarkers and photochemical sensors. The effect on the luminescent property by substituents in the heterocycle and phenyl rings, have been studied as well. In this investigation, series of 3-hydroxymethyl imidazo[1,2-a]pyridines and pyrimidines were synthesized and evaluated in relation to fluorescence emission, based upon the hypothesis that the hydroxymethyl group may act as an enhancer of fluorescence intensity. Results Compounds of both series emitted light in organic solvents dilutions as well as in acidic and alkaline media. Quantitative fluorescence spectroscopy determined that both fused heterocycles fluoresced more intensely than the parent unsubstituted imidazo[1,2-a]azine fluorophore. In particular, 3-hydroxymethyl imidazo[1,2-a]pyridines fluoresced more intensely than 3-hydroxymethyl imidazo[1,2-a]pyrimidines, the latter emitting blue light at longer wavelengths, whereas the former emitted purple light. Conclusion It was concluded that in most cases the hydroxymethyl moiety did act as an enhancer of the fluorescence intensity, however, a comparison made with the fluorescence emitted by 2-aryl imidazo[1,2-a]azines revealed that in some cases the hydroxymethyl substituent decreased the fluorescence intensity.

A fluorescent cassette-based strategy for engineering multiple domain fusion proteins

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Khorchid Ahmad

2003-07-01

Full Text Available Abstract Background The engineering of fusion proteins has become increasingly important and most recently has formed the basis of many biosensors, protein purification systems, and classes of new drugs. Currently, most fusion proteins consist of three or fewer domains, however, more sophisticated designs could easily involve three or more domains. Using traditional subcloning strategies, this requires micromanagement of restriction enzymes sites that results in complex workaround solutions, if any at all. Results Therefore, to aid in the efficient construction of fusion proteins involving multiple domains, we have created a new expression vector that allows us to rapidly generate a library of cassettes. Cassettes have a standard vector structure based on four specific restriction endonuclease sites and using a subtle property of blunt or compatible cohesive end restriction enzymes, they can be fused in any order and number of times. Furthermore, the insertion of PCR products into our expression vector or the recombination of cassettes can be dramatically simplified by screening for the presence or absence of fluorescence. Conclusions Finally, the utility of this new strategy was demonstrated by the creation of basic cassettes for protein targeting to subcellular organelles and for protein purification using multiple affinity tags.

Quantification of epithelial cells in coculture with fibroblasts by fluorescence image analysis.

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Krtolica, Ana; Ortiz de Solorzano, Carlos; Lockett, Stephen; Campisi, Judith

2002-10-01

To demonstrate that senescent fibroblasts stimulate the proliferation and neoplastic transformation of premalignant epithelial cells (Krtolica et al.: Proc Natl Acad Sci USA 98:12072-12077, 2001), we developed methods to quantify the proliferation of epithelial cells cocultured with fibroblasts. We stained epithelial-fibroblast cocultures with the fluorescent DNA-intercalating dye 4,6-diamidino-2-phenylindole (DAPI), or expressed green fluorescent protein (GFP) in the epithelial cells, and then cultured them with fibroblasts. The cocultures were photographed under an inverted microscope with appropriate filters, and the fluorescent images were captured with a digital camera. We modified an image analysis program to selectively recognize the smaller, more intensely fluorescent epithelial cell nuclei in DAPI-stained cultures and used the program to quantify areas with DAPI fluorescence generated by epithelial nuclei or GFP fluorescence generated by epithelial cells in each field. Analysis of the image areas with DAPI and GFP fluorescences produced nearly identical quantification of epithelial cells in coculture with fibroblasts. We confirmed these results by manual counting. In addition, GFP labeling permitted kinetic studies of the same coculture over multiple time points. The image analysis-based quantification method we describe here is an easy and reliable way to monitor cells in coculture and should be useful for a variety of cell biological studies. Copyright 2002 Wiley-Liss, Inc.

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

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

2011-07-10

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

Imaging a Large Sample with Selective Plane Illumination Microscopy Based on Multiple Fluorescent Microsphere Tracking

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Ryu, Inkeon; Kim, Daekeun

2018-04-01

A typical selective plane illumination microscopy (SPIM) image size is basically limited by the field of view, which is a characteristic of the objective lens. If an image larger than the imaging area of the sample is to be obtained, image stitching, which combines step-scanned images into a single panoramic image, is required. However, accurately registering the step-scanned images is very difficult because the SPIM system uses a customized sample mount where uncertainties for the translational and the rotational motions exist. In this paper, an image registration technique based on multiple fluorescent microsphere tracking is proposed in the view of quantifying the constellations and measuring the distances between at least two fluorescent microspheres embedded in the sample. Image stitching results are demonstrated for optically cleared large tissue with various staining methods. Compensation for the effect of the sample rotation that occurs during the translational motion in the sample mount is also discussed.

Morphological spot counting from stacked images for automated analysis of gene copy numbers by fluorescence in situ hybridization.

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Grigoryan, Artyom M; Dougherty, Edward R; Kononen, Juha; Bubendorf, Lukas; Hostetter, Galen; Kallioniemi, Olli

2002-01-01

Fluorescence in situ hybridization (FISH) is a molecular diagnostic technique in which a fluorescent labeled probe hybridizes to a target nucleotide sequence of deoxyribose nucleic acid. Upon excitation, each chromosome containing the target sequence produces a fluorescent signal (spot). Because fluorescent spot counting is tedious and often subjective, automated digital algorithms to count spots are desirable. New technology provides a stack of images on multiple focal planes throughout a tissue sample. Multiple-focal-plane imaging helps overcome the biases and imprecision inherent in single-focal-plane methods. This paper proposes an algorithm for global spot counting in stacked three-dimensional slice FISH images without the necessity of nuclei segmentation. It is designed to work in complex backgrounds, when there are agglomerated nuclei, and in the presence of illumination gradients. It is based on the morphological top-hat transform, which locates intensity spikes on irregular backgrounds. After finding signals in the slice images, the algorithm groups these together to form three-dimensional spots. Filters are employed to separate legitimate spots from fluorescent noise. The algorithm is set in a comprehensive toolbox that provides visualization and analytic facilities. It includes simulation software that allows examination of algorithm performance for various image and algorithm parameter settings, including signal size, signal density, and the number of slices.

Photobleaching and Fluorescence Recovery of RPE Bisretinoids.

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Zhao Liu

Full Text Available The autofluorescence of the retina that originates primarily from lipofuscin fluorophores in retinal pigment epithelial cells, is observed to undergo photobleaching during the acquisition of fundus autofluorescence images. Bisretinoid fluorophores isolated from retinal pigment epithelial cells have the spectral characteristics consistent with their being the source of fundus autofluorescence. Clinically relevant experiments were designed to better understand conditions in the micromilieu of bisretinoid fluorophores that can influence fluorescence efficiencies, photobleaching, and subsequent fluorescence recovery of this fluorophore. The consumption of the bisretinoid A2E due to photooxidation-induced degradation was quantified in solvent systems of variable relative permittivity (formerly called dielectric constant, in micelles, and in phospholipid vesicles of varying composition. Reorganization within biphasic systems was also examined. A2E content was measured by high performance liquid chromatography (HPLC and fluorescence intensity was quantified spectroscopically. As solvent polarity was increased, A2E fluorescent spectra exhibited red-shifted maxima and reduced intensity. A2E was depleted by light irradiation and the loss was more pronounced in less polar solvents, lower concentrations of anionic surfactant, and in gel- versus fluid-ordered phospholipid liposomes. Conditions that permit A2E aggregation promoted photooxidation/photodegradation, while movement of A2E between bisphasic systems was associated with fluorescence recovery after photobleaching. The fluorescence characteristics of A2E are subject to environmental modulation. Photooxidation and photodegradation of bisretinoid can account for fundus autofluorescence photobleaching. Return of fluorescence intensity after photobleaching likely occurs due to redistribution of A2E fractions amongst co-existing heterogeneous microdomains of the lysosomal compartment.

A fluorescence detection of D-penicillamine based on Cu(2+)-induced fluorescence quenching system of protein-stabilized gold nanoclusters.

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Wang, Peng; Li, Bang Lin; Li, Nian Bing; Luo, Hong Qun

2015-01-25

In this contribution, a luminescent gold nanoclusters which were synthesized by bovine serum albumin as novel fluorescent probes were successfully utilized for the determination of D-penicillamine for the first time. Cupric ion was employed to quench the strong fluorescence of the gold nanoclusters, whereas the addition of D-penicillamine caused obvious restoration of fluorescence intensity of the Cu(2+)-gold nanoclusters system. Under optimum conditions, the increment in fluorescence intensity of Cu(2+)-gold nanoclusters system caused by D-penicillamine was linearly proportional to the concentration of D-penicillamine in the range of 2.0×10(-5)-2.39×10(-4) M. The detection limit for D-penicillamine was 5.4×10(-6) M. With the off-on fluorescence signal at 650 nm approaching the near-infrared region, the present sensor for D-penicillamine detection had high sensitivity and low spectral interference. Furthermore, the novel gold nanoclusters-based fluorescent sensor has been applied to the determination of D-penicillamine in real biological samples with satisfactory results. Copyright © 2014 Elsevier B.V. All rights reserved.

Validation of interphase fluorescence in situ hybridization (iFISH for multiple myeloma using CD138 positive cells

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Renata Kiyomi Kishimoto

2016-06-01

Full Text Available ABSTRACT BACKGROUND: Multiple myeloma is a plasma cell neoplasm with acquired genetic abnormalities of clinical and prognostic importance. Multiple myeloma differs from other hematologic malignancies due to a high fraction of low proliferating malignant plasma cells and the paucity of plasma cells in bone marrow aspiration samples, making cytogenetic analysis a challenge. An abnormal karyotype is found in only one-third of patients with multiple myeloma and interphase fluorescence in situ hybridization is the most useful test for studying the chromosomal abnormalities present in almost 90% of cases. However, it is necessary to study the genetic abnormalities in plasma cells after their identification or selection by morphology, immunophenotyping or sorting. Other challenges are the selection of the most informative FISH panel and determining cut-off levels for FISH probes. This study reports the validation of interphase fluorescence in situ hybridization using CD138 positive cells, according to proposed guidelines published by the European Myeloma Network (EMN in 2012. METHOD: Bone marrow samples from patients with multiple myeloma were used to standardize a panel of five probes [1q amplification, 13q14 deletion, 17p deletion, t(4;14, and t(14;16] in CD138+ cells purified by magnetic cell sorting. RESULTS: This test was validated with a low turnaround time and good reproducibility. Five of six samples showed genetic abnormalities. Monosomy/deletion 13 plus t(4;14 were found in two cases. CONCLUSION: This technique together with magnetic cell sorting is effective and can be used in the routine laboratory practice. In addition, magnetic cell sorting provides a pure plasma cell population that allows other molecular and genomic studies.

Helix-coil transition of a four-way DNA junction observed by multiple fluorescence parameters.

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Vámosi, György; Clegg, Robert M

2008-10-16

The thermal denaturation of immobile four-way DNA ("Holliday-") junctions with 17 base pair arms was studied via fluorescence spectroscopic measurements. Two arms of the molecule were labeled at the 5'-end with fluorescein and tetramethylrhodamine, respectively. Melting was monitored by the fluorescence intensity of the dyes, the fluorescence anisotropy of tetramethylrhodamine, and Forster resonance energy transfer (FRET) between fluorescein and rhodamine. To fit the thermal denaturation curves of the four-way junctions, two basic thermodynamic models were tested: (1) all-or-none transitions assuming a molecularity of one, two, or four and (2) a statistical "zipper" model. The all-or-none models correspond to reaction mechanisms assuming that the cooperative melting unit (that is, the structure changing from complete helix to complete coil) consists of (1) one arm, (2) two neighboring arms (which have one continuous strand common to the two arms), or (3) all four arms. In each case, the melting of the cooperative unit takes place in a single step. The tetramolecular reaction model (four-arm melting) yielded unrealistically low van't Hoff enthalpy and entropy values, whereas the monomolecular model (one-arm melting) resulted in a poor fit to the experimental data. The all-or-none bimolecular (two neighboring arm model) fit gave intermediate standard enthalpy change (Delta H) values between those expected for the melting of a duplex with a total length between the helix lengths of one and two arms (17 and 34 base pairs). Simulations according to the zipper model fit the experimental curves best when the length of the simulated duplex was assumed to be 34 base pairs, the length of a single strand. This suggests that the most important parameter determining the melting behavior of the molecule is the end-to-end distance of the strands (34 bases) rather than the length of the individual arms (17 base pairs) and that the equilibrium concentration of partially denatured

Fluorescence uranium determination

International Nuclear Information System (INIS)

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

1960-01-01

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

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

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Zhong, Xin; Wang, Xinwei; Zhou, Yan

2018-01-01

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

A label-free, fluorescence based assay for microarray

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Niu, Sanjun

intensity of excitation light. The fluorescence contrast is used to quantify the amount of probe-target hybridization. A mathematical model that considers multiple reflections and scattering is developed to explain the mechanism of the fluorescence contrast which depends on the thickness of the PS film. Scattering is the dominant factor that contributes to the contrast. The potential of this assay to detect single nucleotide polymorphism is also tested.

Enhanced speed in fluorescence imaging using beat frequency multiplexing

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Mikami, Hideharu; Kobayashi, Hirofumi; Wang, Yisen; Hamad, Syed; Ozeki, Yasuyuki; Goda, Keisuke

2016-03-01

Fluorescence imaging using radiofrequency-tagged emission (FIRE) is an emerging technique that enables higher imaging speed (namely, temporal resolution) in fluorescence microscopy compared to conventional fluorescence imaging techniques such as confocal microscopy and wide-field microscopy. It works based on the principle that it uses multiple intensity-modulated fields in an interferometric setup as excitation fields and applies frequency-division multiplexing to fluorescence signals. Unfortunately, despite its high potential, FIRE has limited imaging speed due to two practical limitations: signal bandwidth and signal detection efficiency. The signal bandwidth is limited by that of an acousto-optic deflector (AOD) employed in the setup, which is typically 100-200 MHz for the spectral range of fluorescence excitation (400-600 nm). The signal detection efficiency is limited by poor spatial mode-matching between two interfering fields to produce a modulated excitation field. Here we present a method to overcome these limitations and thus to achieve higher imaging speed than the prior version of FIRE. Our method achieves an increase in signal bandwidth by a factor of two and nearly optimal mode matching, which enables the imaging speed limited by the lifetime of the target fluorophore rather than the imaging system itself. The higher bandwidth and better signal detection efficiency work synergistically because higher bandwidth requires higher signal levels to avoid the contribution of shot noise and amplifier noise to the fluorescence signal. Due to its unprecedentedly high-speed performance, our method has a wide variety of applications in cancer detection, drug discovery, and regenerative medicine.

Laser-excited fluorescence for measuring atmospheric pollution

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Menzies, R. T.

1975-01-01

System measures amount of given pollutant at specific location. Infrared laser aimed at location has wavelength that will cause molecules of pollutant to fluoresce. Detector separates fluorescence from other radiation and measures its intensity to indicate concentration of pollutant.

Angular shaping of fluorescence from synthetic opal-based photonic crystal.

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Boiko, Vitalii; Dovbeshko, Galyna; Dolgov, Leonid; Kiisk, Valter; Sildos, Ilmo; Loot, Ardi; Gorelik, Vladimir

2015-01-01

Spectral, angular, and temporal distributions of fluorescence as well as specular reflection were investigated for silica-based artificial opals. Periodic arrangement of nanosized silica globules in the opal causes a specific dip in the defect-related fluorescence spectra and a peak in the reflectance spectrum. The spectral position of the dip coincides with the photonic stop band. The latter is dependent on the size of silica globules and the angle of observation. The spectral shape and intensity of defect-related fluorescence can be controlled by variation of detection angle. Fluorescence intensity increases up to two times at the edges of the spectral dip. Partial photobleaching of fluorescence was observed. Photonic origin of the observed effects is discussed.

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

International Nuclear Information System (INIS)

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

2007-01-01

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

Fluorescence Stability of Mercaptopropionic Acid Capped Cadmium Telluride Quantum Dots in Various Biochemical Buffers.

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Borse, Vivek; Kashikar, Adisha; Srivastava, Rohit

2018-04-01

Quantum dots are the semiconductor nanocrystals having unique optical and electronic properties. Quantum dots are category of fluorescent labels utilized for biological tagging, biosensing, bioassays, bioimaging and in vivo imaging as they exhibit very small size, signal brightness, photostability, tuning of light emission range, longer photoluminescence decay time as compared to organic dyes. In this work, we have synthesized and characterized mercaptopropionic acid capped cadmium telluride quantum dots (MPA-CdTe QDs) using hydrothermal method. The study further reports fluorescence intensity stability of quantum dots suspended in different buffers of varying concentration (1-100 mM), stored at various photophysical conditions. Fluorescence intensity values were reduced with increase in buffer concentration. When the samples were stored at room temperature in ambient light condition the quantum dots suspended in different buffers lost the fluorescence intensity after day 15 (except TRIS II). Fluorescence intensity values were found stable for more than 30 days when the samples were stored in dark condition. Samples stored in refrigerator displayed modest fluorescence intensity even after 300 days of storage. Thus, storage of MPA-CdTe QDs in refrigerator may be the suitable choice to maintain its fluorescence stability for longer time for further application.

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

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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.

Fabrication of fluorescent chitosan-containing microcapsules

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Zhang R.

2013-08-01

Full Text Available Intense emission peaks of Eu(DBM3Phen (DBM and Phen are dibenzoylmethane and 1,10-phenanthroline, respectively in the microcapsules containing molecules of quaternary ammonium chitosan (QACS and sodium alginate are observed. The microcapsules are assembled by using CaCO3 particles as template cores by the layer-by-layer (LbL technique. Observation of microcapsules by the fluorescence mode and the transmission mode in the confocal laser scanning microscopy shows that the microcapsules are intact after core decomposition. Fluorescence under ultraviolet irradiation comes directly from the Eu(DBM3Phen. Homogeneous assembly of Eu(DBM3Phen can be deduced due to the homogeneous fluorescence of the microcapsules in the fluorescence micrographs. The microcapsules show adherence to solid substrates due to large quantities of hydroxyl groups of QACS. AFM measurements of dried hollow microcapsules with only 4 bilayers of (CS/SA fabricated with Eu(DBM3Phen show the intact shell with a thickness of 3.0 nm. Regarding the biocompatible natural polysaccharides and the intense fluorescence emission, the microcapsules in this work might be of great importance in potential application in drug delivery and bioassay.

Fuel Mix Impacts from Transportation Fuel Carbon Intensity Standards in Multiple Jurisdictions

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Witcover, J.

2017-12-01

Fuel carbon intensity standards have emerged as an important policy in jurisdictions looking to target transportation greenhouse gas (GHG) emissions for reduction. A carbon intensity standard rates transportation fuels based on analysis of lifecycle GHG emissions, and uses a system of deficits and tradable, bankable credits to reward increased use of fuels with lower carbon intensity ratings while disincentivizing use of fuels with higher carbon intensity ratings such as conventional fossil fuels. Jurisdictions with carbon intensity standards now in effect include California, Oregon, and British Columbia, all requiring 10% reductions in carbon intensity of the transport fuel pool over a 10-year period. The states and province have committed to grow demand for low carbon fuels in the region as part of collaboration on climate change policies. Canada is developing a carbon intensity standard with broader coverage, for fuels used in transport, industry, and buildings. This study shows a changing fuel mix in affected jurisdictions under the policy in terms of shifting contribution of transportation energy from alternative fuels and trends in shares of particular fuel pathways. It contrasts program designs across the jurisdictions with the policy, highlights the opportunities and challenges these pose for the alternative fuel market, and discusses the impact of having multiple policies alongside federal renewable fuel standards and sometimes local carbon pricing regimes. The results show how the market has responded thus far to a policy that incentivizes carbon saving anywhere along the supply chain at lowest cost, in ways that diverged from a priori policy expectations. Lessons for the policies moving forward are discussed.

Fluorescence imaging of soybean flavonol isolines

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

1998-07-01

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

Holistic needs assessment in advanced, intensively treated multiple myeloma patients.

Science.gov (United States)

Boland, E G; Boland, J W; Ezaydi, Y; Greenfield, D M; Ahmedzai, S H; Snowden, J A

2014-10-01

It is recommended that patients with multiple myeloma should be assessed for unmet holistic needs at key times in their disease trajectory. The aim of this exploratory study was to characterise the holistic needs of advanced, intensively treated multiple myeloma using a structured assessment tool. Patients with multiple myeloma who had undergone a haematopoietic stem cell transplantation and subsequent treatment for at least one episode of progressive disease but were in stable plateau phase were included in the study. Patients' holistic needs were assessed using the self-reporting tool, Sheffield Profile for Assessment and Referral for Care (SPARC). Thirty-two patients with a median age of 60 years at assessment and a median of 5.5 years from diagnosis were recruited. Using the SPARC, half of the patients reported tiredness as 'quite a bit/very much,' while one third complained that daytime somnolence and insomnia were 'quite a bit/very much.' Forty-four percent of patients reported pain. One third of patients were bothered and distressed by the side effects from their treatment and were worried about long-term effects of their treatment. Thirty-one percent of patients felt that the effect of their condition had an impact on their sexual life, and 40 % were worried about the effect that their illness was having on their family or other people. This is the first study to use a self-reported holistic needs assessment tool in multiple myeloma. A multidimensional structured questionnaire like the SPARC could provide a useful first step in the effective delivery of supportive and palliative care for patients with multiple myeloma.

Accurate study of FosPeg® distribution in a mouse model using fluorescence imaging technique and fluorescence white monte carlo simulations

DEFF Research Database (Denmark)

Xie, Haiyan; Liu, Haichun; Svenmarker, Pontus

2010-01-01

Fluorescence imaging is used for quantitative in vivo assessment of drug concentration. Light attenuation in tissue is compensated for through Monte-Carlo simulations. The intrinsic fluorescence intensity, directly proportional to the drug concentration, could be obtained....

Experimental assessment of fluorescence microscopy signal enhancement by stimulated emission

Science.gov (United States)

Dake, Fumihiro; Yazawa, Hiroki

2017-10-01

The quantity of photons generated during fluorescence microscopy is principally determined by the quantum yield of the fluorescence dyes and the optical power of the excitation beam. However, even though low quantum yields can produce poor images, it is challenging to tune this parameter, while increasing the power of the excitation beam often results in photodamage. Here, we propose the use of stimulated emission (SE) as a means of enhancing both the signal intensity and signal-to-noise ratio during confocal fluorescence microscopy. This work experimentally confirmed that both these factors can be enhanced by SE radiation, through generating a greater number of photons than are associated with the standard fluorescence signal. We also propose the concept of stimulated emission enhancing fluorescence (SEEF) microscopy, which employs both the SE and fluorescence signals, and demonstrate that the intensity of an SEEF signal is greater than those of the individual SE and fluorescence signals.

Fluorescence detection of dental calculus

International Nuclear Information System (INIS)

Gonchukov, S; Sukhinina, A; Vdovin, Yu; Biryukova, T

2010-01-01

This work is devoted to the optimization of fluorescence dental calculus diagnostics in optical spectrum. The optimal wavelengths for fluorescence excitation and registration are determined. Two spectral ranges 620 – 645 nm and 340 – 370 nm are the most convenient for supra- and subgingival calculus determination. The simple implementation of differential method free from the necessity of spectrometer using was investigated. Calculus detection reliability in the case of simple implementation is higher than in the case of spectra analysis at optimal wavelengths. The use of modulated excitation light and narrowband detection of informative signal allows us to decrease essentially its diagnostic intensity even in comparison with intensity of the low level laser dental therapy

Fluorescence detection of dental calculus

Science.gov (United States)

Gonchukov, S.; Biryukova, T.; Sukhinina, A.; Vdovin, Yu

2010-11-01

This work is devoted to the optimization of fluorescence dental calculus diagnostics in optical spectrum. The optimal wavelengths for fluorescence excitation and registration are determined. Two spectral ranges 620 - 645 nm and 340 - 370 nm are the most convenient for supra- and subgingival calculus determination. The simple implementation of differential method free from the necessity of spectrometer using was investigated. Calculus detection reliability in the case of simple implementation is higher than in the case of spectra analysis at optimal wavelengths. The use of modulated excitation light and narrowband detection of informative signal allows us to decrease essentially its diagnostic intensity even in comparison with intensity of the low level laser dental therapy.

Application of fluorescence spectroscopy and imaging in the detection of a photosensitizer in photodynamic therapy

Science.gov (United States)

Zang, Lixin; Zhao, Huimin; Zhang, Zhiguo; Cao, Wenwu

2017-02-01

Photodynamic therapy (PDT) is currently an advanced optical technology in medical applications. However, the application of PDT is limited by the detection of photosensitizers. This work focuses on the application of fluorescence spectroscopy and imaging in the detection of an effective photosenzitizer, hematoporphyrin monomethyl ether (HMME). Optical properties of HMME were measured and analyzed based on its absorption and fluorescence spectra. The production mechanism of its fluorescence emission was analyzed. The detection device for HMME based on fluorescence spectroscopy was designed. Ratiometric method was applied to eliminate the influence of intensity change of excitation sources, fluctuates of excitation sources and photo detectors, and background emissions. The detection limit of this device is 6 μg/L, and it was successfully applied to the diagnosis of the metabolism of HMME in the esophageal cancer cells. To overcome the limitation of the point measurement using fluorescence spectroscopy, a two-dimensional (2D) fluorescence imaging system was established. The algorithm of the 2D fluorescence imaging system is deduced according to the fluorescence ratiometric method using bandpass filters. The method of multiple pixel point addition (MPPA) was used to eliminate fluctuates of signals. Using the method of MPPA, SNR was improved by about 30 times. The detection limit of this imaging system is 1.9 μg/L. Our systems can be used in the detection of porphyrins to improve the PDT effect.

Synthesis and fluorescence properties of some difluoroboron β-diketonate complexes and composite containing PMMA

Science.gov (United States)

Xing, Dongye; Hou, Yanjun; Niu, Haijun

2018-03-01

A series of difluoroboron β-diketonate complexes, containing the indon-β-diketonate ligand carrying methyl or methoxyl substituents was synthesized. The crystal structures of the complexes were confirmed by single crystal X-ray diffraction studies. The fluorescence properties of compounds were studied in solution state, solid state and on PMMA polymer matrix. The photophysical data of compounds 2a-2d exhibited strong fluorescence and photostability under the ultraviolet light (Hg lamp). The complex 2b showed higher fluorescence intensity in solution state as compared to other complexes of the series. The complexes 2c and 2d showed higher fluorescence intensity in the solid state, which are ascribed to the stronger π-π interactions between ligands in the solid state. The introduction of methoxyl or methyl groups on the benzene rings enhanced the absorption intensity, emission intensity, quantum yields and fluorescence lifetimes due to their electron-donating nature. Furthermore, the complex 2b was doped into the PMMA to produce hybrid materials, where the PMMA matrix acted as sensitizer for the central boron ion to enhance the fluorescence emission intensity and quantum yields.

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

Science.gov (United States)

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

2017-09-01

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

Nine New Fluorescent Probes

Science.gov (United States)

Lin, Tsung-I.; Jovanovic, Misa V.; Dowben, Robert M.

1989-06-01

Absorption and fluorescence spectroscopic studies are reported here for nine new fluorescent probes recently synthesized in our laboratories: four pyrene derivatives with substituents of (i) 1,3-diacetoxy-6,8-dichlorosulfonyl, (ii) 1,3-dihydroxy-6,8-disodiumsulfonate, (iii) 1,3-disodiumsulfonate, and (iv) l-ethoxy-3,6,8-trisodiumsulfonate groups, and five [7-julolidino] coumarin derivatives with substituents of (v) 3-carboxylate-4-methyl, (vi) 3- methylcarboxylate, (vii) 3-acetate-4-methyl, (viii) 3-propionate-4-methyl, and (ix) 3-sulfonate-4-methyl groups. Pyrene compounds i and ii and coumarin compounds v and vi exhibit interesting absorbance and fluorescence properties: their absorption maxima are red shifted compared to the parent compound to the blue-green region, and the band width broadens considerably. All four blue-absorbing dyes fluoresce intensely in the green region, and the two pyrene compounds emit at such long wavelengths without formation of excimers. The fluorescence properties of these compounds are quite environment-sensitive: considerable spectral shifts and fluorescence intensity changes have been observed in the pH range from 3 to 10 and in a wide variety of polar and hydrophobic solvents with vastly different dielectric constants. The high extinction and fluorescence quantum yield of these probes make them ideal fluorescent labeling reagents for proteins, antibodies, nucleic acids, and cellular organelles. The pH and hydrophobicity-dependent fluorescence changes can be utilized as optical pH and/or hydrophobicity indicators for mapping environmental difference in various cellular components in a single cell. Since all nine probes absorb in the UV, but emit at different wavelengths in the visible, these two groups of compounds offer an advantage of utilizing a single monochromatic light source (e.g., a nitrogen laser) to achieve multi-wavelength detection for flow cytometry application. As a first step to explore potential application in

Multidimensional fluorescence microscopy of multiple organelles in Arabidopsis seedlings

Directory of Open Access Journals (Sweden)

Morales Andrea

2008-05-01

Full Text Available Abstract Background The isolation of green fluorescent protein (GFP and the development of spectral variants over the past decade have begun to reveal the dynamic nature of protein trafficking and organelle motility. In planta analyses of this dynamic process have typically been limited to only two organelles or proteins at a time in only a few cell types. Results We generated a transgenic Arabidopsis plant that contains four spectrally different fluorescent proteins. Nuclei, plastids, mitochondria and plasma membranes were genetically tagged with cyan, red, yellow and green fluorescent proteins, respectively. In addition, methods to track nuclei, mitochondria and chloroplasts and quantify the interaction between these organelles at a submicron resolution were developed. These analyzes revealed that N-ethylmaleimide disrupts nuclear-mitochondrial but not nuclear-plastids interactions in root epidermal cells of live Arabidopsis seedlings. Conclusion We developed a tool and associated methods for analyzing the complex dynamic of organelle-organelle interactions in real time in planta. Homozygous transgenic Arabidopsis (Kaleidocell is available through Arabidopsis Biological Resource Center.

Multiple ionization dynamics of molecules in intense laser fields

International Nuclear Information System (INIS)

Ichimura, Atsushi; Ohyama-Yamaguchi, Tomoko

2005-01-01

A classical field-ionization model is developed for sequential multiple ionization of diatomic and linear triatomic molecules exposed to intense (∼ 10 15 W/cm 2 ) laser fields. The distance R ion of Coulomb explosion is calculated for a combination of fragment charges, by considering nonadiabatic excitation followed by field ionization associated with the inner and outer saddle points. For diatomic molecules (N 2 , NO, and I 2 ), the model explains behaviors observed in experiments, as R ion (21→31) ion (21→22) between competing charge-asymmetric and symmetric channels, and even-odd fluctuation along a principal pathway. For a triatomic molecule CO 2 , a comparison of the model with an experiment suggests that charge-symmetric (or nearly symmetric) channels are dominantly populated. (author)

Construction of In Vivo Fluorescent Imaging of Echinococcus granulosus in a Mouse Model.

Science.gov (United States)

Wang, Sibo; Yang, Tao; Zhang, Xuyong; Xia, Jie; Guo, Jun; Wang, Xiaoyi; Hou, Jixue; Zhang, Hongwei; Chen, Xueling; Wu, Xiangwei

2016-06-01

Human hydatid disease (cystic echinococcosis, CE) is a chronic parasitic infection caused by the larval stage of the cestode Echinococcus granulosus. As the disease mainly affects the liver, approximately 70% of all identified CE cases are detected in this organ. Optical molecular imaging (OMI), a noninvasive imaging technique, has never been used in vivo with the specific molecular markers of CE. Thus, we aimed to construct an in vivo fluorescent imaging mouse model of CE to locate and quantify the presence of the parasites within the liver noninvasively. Drug-treated protoscolices were monitored after marking by JC-1 dye in in vitro and in vivo studies. This work describes for the first time the successful construction of an in vivo model of E. granulosus in a small living experimental animal to achieve dynamic monitoring and observation of multiple time points of the infection course. Using this model, we quantified and analyzed labeled protoscolices based on the intensities of their red and green fluorescence. Interestingly, the ratio of red to green fluorescence intensity not only revealed the location of protoscolices but also determined the viability of the parasites in vivo and in vivo tests. The noninvasive imaging model proposed in this work will be further studied for long-term detection and observation and may potentially be widely utilized in susceptibility testing and therapeutic effect evaluation.

Fluorescent multiplex cell flow systems and methods

KAUST Repository

Merzaban, Jasmeen

2017-06-01

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

Detection of Counterfeit Tequila by Fluorescence Spectroscopy

Directory of Open Access Journals (Sweden)

José Manuel de la Rosa Vázquez

2015-01-01

Full Text Available An ultraviolet (UV light induced fluorescence study to discriminate fake tequila from genuine ones is presented. A portable homemade system based on four light emitting diodes (LEDs from 255 to 405 nm and a miniature spectrometer was used. It has been shown that unlike fake and silver tequila, which produce weak fluorescence signal, genuine mixed, rested, and aged tequilas show high fluorescence emission in the range from 400 to 750 nm. The fluorescence intensity grows with aging in 100% agave tequila. Such fluorescence differences can even be observed with naked eyes. The presented results demonstrate that the fluorescence measurement could be a good method to detect counterfeit tequila.

Fluorescence in situ hybridization detection of cytogenetic abnormalities and prognosis in multiple myeloma

Directory of Open Access Journals (Sweden)

Zhou Xu

2015-01-01

Full Text Available We evaluated the prognosis of patients with newly diagnosed multiple myeloma (MM and attempted to find a suitable treatment strategy for them. Interphase fluorescence in situ hybridization (FISH detection was performed on 57 patients with MM. The following probes: IgH, p53, 1q21, RB1, and D13S319 specific for the rearrangements of 14q32, 17p13, 1q21 and 13q14 were used. Fluorescent hybridization signals were observed using an Olympus BX60 epifluorescence microscope equipped with filters for detecting fluoroisothiocyanate (FITC, Texas red, and 4'-6-Diamidino-2-phenylindole (DAPI. Triple color clone-specific images were captured using a Quips XL genetic workstation. The mortalities in patients with moderate prognosis (66.7% and poor prognosis (50% were significantly higher compared with that in patients with good prognosis (15%, P<0.05. All the patients in good and moderate prognosis groups achieved complete remission (CR/very good partial remission (VGPR/partial remission (PR, whereas only half of the cases in the poor prognosis group reached this level. Patients 2 supported by autologous hematopoietic stem-cell transplantation presented CR/PR and long survival. For those with poor prognosis, a proper therapeutic regimen and timely transplantation, especially tandem transplantation, was necessary due to the rapid progression and complications.

Direct spectrometry: a new alternative for measuring the fluorescence of composite resins and dental tissues.

Science.gov (United States)

da Silva, Tm; de Oliveira, Hpm; Severino, D; Balducci, I; Huhtala, Mfrl; Gonçalves, Sep

2014-01-01

The aim of this study was to evaluate the fluorescence intensity of different composite resins and compare those values with the fluorescence intensity of dental tissues. Different composite resins were used to make 10 discs (2 mm in depth and 4 mm in diameter) of each brand, divided into groups: 1) Z (Filtek Z350, 3M ESPE), 2) ES (Esthet-X, Dentsply), 3) A (Amelogen Plus, Ultradent), 4) DVS (Durafill-VS, Heraeus Kulzer) with 2 mm composite resin for enamel (A2), 5) OES ([Esthet-X] opaque-OA [1 mm] + enamel-A2 [1 mm]); 6) ODVSI ([Charisma-Opal/Durafill-VSI], opaque-OM (1 mm) + translucent [1mm]), and 7) DVSI ([Durafill- VSI] translucent [2 mm]). Dental tissue specimens were obtained from human anterior teeth cut in a mesiodistal direction to obtain enamel, dentin, and enamel/dentin samples (2 mm). The fluorescence intensity of specimens was directly measured using an optic fiber associated with a spectrometer (Ocean Optics USB 4000) and recorded in graphic form (Origin 8.0 program). Data were submitted to statistical analysis using Dunnet, Tukey, and Kruskall-Wallis tests. Light absorption of the composite resins was obtained in a spectral range from 250 to 450 nm, and that of dental tissues was between 250 and 300 nm. All composite resins were excited at 398 nm and exhibited maximum emissions of around 485 nm. Fluorescence intensity values for all of the resins showed statistically significant differences (measured in arbitrary units [AUs]), with the exception of groups Z and DVS. Group DVSI had the highest fluorescence intensity values (13539 AU), followed by ODVS (10440 AU), DVS (10146 AU), ES (3946 AU), OES (3841 AU), A (3540 AU), and Z (1146 AU). The fluorescence intensity values for the composite resins differed statistically from those of dental tissues (E=1380 AU; D=6262 AU; E/D=3251 AU). The opacity interfered with fluorescence intensity, and group Z demonstrated fluorescence intensity values closest to that of tooth enamel. It is concluded that the

Reliable Assessment and Quantification of the Fluorescence-Labeled Antisense Oligonucleotides In Vivo

Directory of Open Access Journals (Sweden)

Maria Chiara Munisso

2014-01-01

Full Text Available The availability of fluorescent dyes and the advances in the optical systems for in vivo imaging have stimulated an increasing interest in developing new methodologies to study and quantify the biodistribution of labeled agents. However, despite these great achievements, we are facing significant challenges in determining if the observed fluorescence does correspond to the quantity of the dye in the tissues. In fact, although the far-red and near-infrared lights can propagate through several centimetres of tissue, they diffuse within a few millimetres as consequence of the elastic scattering of photons. In addition, when dye-labeled oligonucleotides form stable complex with cationic carriers, a large change in the fluorescence intensity of the dye is observed. Therefore, the measured fluorescence intensity is altered by the tissue heterogeneity and by the fluctuation of dye intensity. Hence, in this study a quantification strategy for fluorescence-labeled oligonucleotides was developed to solve these disadvantageous effects. Our results proved that upon efficient homogenization and dilution with chaotropic agents, such as guanidinium thiocyanate, it is possible to achieve a complete fluorescence intensity recovery. Furthermore, we demonstrated that this method has the advantage of good sensitivity and reproducibility, as well as easy handling of the tissue samples.

Workflow for high-content, individual cell quantification of fluorescent markers from universal microscope data, supported by open source software.

Science.gov (United States)

Stockwell, Simon R; Mittnacht, Sibylle

2014-12-16

Advances in understanding the control mechanisms governing the behavior of cells in adherent mammalian tissue culture models are becoming increasingly dependent on modes of single-cell analysis. Methods which deliver composite data reflecting the mean values of biomarkers from cell populations risk losing subpopulation dynamics that reflect the heterogeneity of the studied biological system. In keeping with this, traditional approaches are being replaced by, or supported with, more sophisticated forms of cellular assay developed to allow assessment by high-content microscopy. These assays potentially generate large numbers of images of fluorescent biomarkers, which enabled by accompanying proprietary software packages, allows for multi-parametric measurements per cell. However, the relatively high capital costs and overspecialization of many of these devices have prevented their accessibility to many investigators. Described here is a universally applicable workflow for the quantification of multiple fluorescent marker intensities from specific subcellular regions of individual cells suitable for use with images from most fluorescent microscopes. Key to this workflow is the implementation of the freely available Cell Profiler software(1) to distinguish individual cells in these images, segment them into defined subcellular regions and deliver fluorescence marker intensity values specific to these regions. The extraction of individual cell intensity values from image data is the central purpose of this workflow and will be illustrated with the analysis of control data from a siRNA screen for G1 checkpoint regulators in adherent human cells. However, the workflow presented here can be applied to analysis of data from other means of cell perturbation (e.g., compound screens) and other forms of fluorescence based cellular markers and thus should be useful for a wide range of laboratories.

The exploration of the characteristics of the hyperglycemia serum fluorescence spectrum

Science.gov (United States)

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.

[Intensive care treatment of traumatic brain injury in multiple trauma patients : Decision making for complex pathophysiology].

Science.gov (United States)

Trimmel, H; Herzer, G; Schöchl, H; Voelckel, W G

2017-09-01

Traumatic brain injury (TBI) and hemorrhagic shock due to uncontrolled bleeding are the major causes of death after severe trauma. Mortality rates are threefold higher in patients suffering from multiple injuries and additionally TBI. Factors known to impair outcome after TBI, namely hypotension, hypoxia, hypercapnia, acidosis, coagulopathy and hypothermia are aggravated by the extent and severity of extracerebral injuries. The mainstays of TBI intensive care may be, at least temporarily, contradictory to the trauma care concept for multiple trauma patients. In particular, achieving normotension in uncontrolled bleeding situations, maintenance of normocapnia in traumatic lung injury and thromboembolic prophylaxis are prone to discussion. Due to an ongoing uncertainty about the definition of normotensive blood pressure values, a cerebral perfusion pressure-guided cardiovascular management is of key importance. In contrast, there is no doubt that early goal directed coagulation management improves outcome in patients with TBI and multiple trauma. The timing of subsequent surgical interventions must be based on the development of TBI pathology; therefore, intensive care of multiple trauma patients with TBI requires an ongoing and close cooperation between intensivists and trauma surgeons in order to individualize patient care.

Measurement of cell volume changes by fluorescence self-quenching

DEFF Research Database (Denmark)

Hamann, Steffen; Kiilgaard, J.F.; Litman, Thomas

2002-01-01

At high concentrations, certain fluorophores undergo self-quenching, i.e., fluorescence intensity decreases with increasing fluorophore concentration. Accordingly, the self-quenching properties can be used for measuring water volume changes in lipid vesicles. In cells, quantitative determination...... concentrations of the fluorophore calcein suitable for measurement of changes in cell water volume by self-quenching. The relationship between calcein fluorescence intensity, when excited at 490 nm (its excitation maximum), and calcein concentration was investigated in vitro and in various cultured cell types...... to a decrease in calcein fluorescence with high signal-to-noise ratio (>15). Similar results were obtained with the fluorophore BCECF when excited at its isosbestic wavelength (436 nm). The present results demonstrate the usefulness of fluorescence self-quenching to measure rapid changes in cell water volume....

Optimization of mNeonGreen for Homo sapiens increases its fluorescent intensity in mammalian cells.

Science.gov (United States)

Tanida-Miyake, Emiko; Koike, Masato; Uchiyama, Yasuo; Tanida, Isei

2018-01-01

Green fluorescent protein (GFP) is tremendously useful for investigating many cellular and intracellular events. The monomeric GFP mNeonGreen is about 3- to 5-times brighter than GFP and monomeric enhanced GFP and shows high photostability. The maturation half-time of mNeonGreen is about 3-fold faster than that of monomeric enhanced GFP. However, the cDNA sequence encoding mNeonGreen contains some codons that are rarely used in Homo sapiens. For better expression of mNeonGreen in human cells, we synthesized a human-optimized cDNA encoding mNeonGreen and generated an expression plasmid for humanized mNeonGreen under the control of the cytomegalovirus promoter. The resultant plasmid was introduced into HEK293 cells. The fluorescent intensity of humanized mNeonGreen was about 1.4-fold higher than that of the original mNeonGreen. The humanized mNeonGreen with a mitochondria-targeting signal showed mitochondrial distribution of mNeonGreen. We further generated an expression vector of humanized mNeonGreen with 3xFLAG tags at its carboxyl terminus as these tags are useful for immunological analyses. The 3xFLAG-tagged mNeonGreen was recognized well with an anti-FLAG-M2 antibody. These plasmids for the expression of humanized mNeonGreen and mNeonGreen-3xFLAG are useful tools for biological studies in mammalian cells using mNeonGreen.

Surface plasmon-enhanced molecular fluorescence induced by gold nanostructures

International Nuclear Information System (INIS)

Teng, Y.; Ueno, K.; Shi, X.; Aoyo, D.; Misawa, H.; Qiu, J.

2012-01-01

The authors report on surface plasmon-enhanced fluorescence of Eosin Y molecules induced by gold nanostructures. Al 2 O 3 films deposited by atomic layer deposition with sub-nanometer resolution were used as the spacer layer to control the distance between molecules and the gold surface. As the thickness of the Al 2 O 3 film increased, the fluorescence intensity first increased and then decreased. The highest enhancement factor is achieved with a 1 nm Al 2 O 3 film. However, the trend for the fluorescence lifetime is the opposite. It first decreased and then increased. The changes in the fluorescence quantum yield were also calculated. The yield shows a similar trend to the fluorescence intensity. The competition between the surface plasmon-induced increase in the radiative decay rate and the gold-induced fluorescence quenching is responsible for the observed phenomenon. In addition, this competition strongly depends on the thickness of the spacer layer between Eosin Y molecules and the gold surface. (Copyright copyright 2012 WILEY-VCH Verlag GmbH and Co. KGaA, Weinheim)

Confocal fluorescence techniques in industrial application

Science.gov (United States)

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

2003-06-01

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

Noninvasive control of the transport function of fluorescent coloured liposomal nanoparticles

Science.gov (United States)

Stelmashchuk, O.; Zherebtsov, E.; Zherebtsova, A.; Kuznetsova, E.; Vinokurov, A.; Dunaev, A.; Mamoshin, A.; Snimshchikova, I.; Borsukov, A.; Bykov, A.; Meglinski, I.

2017-06-01

The use of liposomal nanoparticles with an incorporated active substance is an innovative and promising approach to diagnostics and therapy. The application of liposomal nanoparticle-based drugs allows for targeted localized delivery, overcomes the natural barriers within the body effectively, and minimizes possible side effects. Liposomes are able to contain a variety of ingredients with practically no limitations to their chemical composition, chemical properties, or size of constituent molecules. This study evaluated the ability to control the passage of fluorescent dye-filled liposomes through the intestinal mucosal barrier after oral administration. For this purpose, the increase in transcutaneous registered fluorescence from tetrabromofluorescein dye was recorded and analysed. Fluorescence intensity was measured at the proximal end of the tail of an animal model after oral administration of the liposomes. Measurements were taken at the excitation wavelengths of 365 and 450 nm. The fluorescence intensity in the group treated with the fluorescent contrast agent encapsulated in liposomal particles increased 140% of the initial level, but in the group treated with pure contrast agent, the increase in detected fluorescence intensity did not exceed 110%. Mice that received empty liposomes as well as the control group did not demonstrate statistically significant changes in fluorescence intensity. A potential application of our results is an express laser optical method of monitoring the transport of orally administered liposomal particles. The results can be used to help create new optical tools for use in the development of new drugs and in high-throughput screening used during their testing.

Multiphoton fluorescence lifetime imaging of chemotherapy distribution in solid tumors

Science.gov (United States)

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

2017-11-01

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

A PDMS-based cylindrical hybrid lens for enhanced fluorescence detection in microfluidic systems.

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Lin, Bor-Shyh; Yang, Yu-Ching; Ho, Chong-Yi; Yang, Han-Yu; Wang, Hsiang-Yu

2014-02-13

Microfluidic systems based on fluorescence detection have been developed and applied for many biological and chemical applications. Because of the tiny amount of sample in the system; the induced fluorescence can be weak. Therefore, most microfluidic systems deploy multiple optical components or sophisticated equipment to enhance the efficiency of fluorescence detection. However, these strategies encounter common issues of complex manufacturing processes and high costs. In this study; a miniature, cylindrical and hybrid lens made of polydimethylsiloxane (PDMS) to improve the fluorescence detection in microfluidic systems is proposed. The hybrid lens integrates a laser focusing lens and a fluorescence collecting lens to achieve dual functions and simplify optical setup. Moreover, PDMS has advantages of low-cost and straightforward fabrication compared with conventional optical components. The performance of the proposed lens is first examined with two fluorescent dyes and the results show that the lens provides satisfactory enhancement for fluorescence detection of Rhodamine 6G and Nile Red. The overall increments in collected fluorescence signal and detection sensitivity are more than 220% of those without lens, and the detection limits of Rhodamine 6G and Nile red are lowered to 0.01 μg/mL and 0.05 μg/mL, respectively. The hybrid lens is further applied to the detection of Nile red-labeled Chlorella vulgaris cells and it increases both signal intensity and detection sensitivity by more than 520%. The proposed hybrid lens also dramatically reduces the variation in detected signal caused by the deviation in incident angle of excitation light.

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

Directory of Open Access Journals (Sweden)

Gregor P. C. Drummen

2012-11-01

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

Seeing the electroporative uptake of cell-membrane impermeable fluorescent molecules and nanoparticles

Science.gov (United States)

Kim, Kisoo; Kim, Jeong Ah; Lee, Soon-Geul; Lee, Won Gu

2012-07-01

This paper presents direct visualization of uptake directionality for cell-membrane impermeant fluorescent molecules and fluorescence-doped nanoparticles at a single-cell level during electroporation. To observe directly the uptake direction, we used microchannel-type electroporation that can generate a relatively symmetric and uniform electric field. For all the image frames during electroporation, fluorescence intensities that occurred at cell membranes in both uptake directions toward the electrodes have been sequentially recorded and quantitatively analyzed pixel by pixel. In our experiments, we found that fluorescent molecules, even not labeled to target biomolecules, had their own uptake direction with different intensities. It is also observed that the uptake intensity toward the cell membrane had a maximal value at a certain electric voltage, not at the highest value of voltages applied. The results also imply that the uptake direction of fluorescence-doped nanoparticles can be determined by a net surface charge of uptake materials and sizes in the electroporative environments. In summary, we performed a quantitative screening and direct visualization of uptake directionality for a set of fluorescent molecules and fluorescence-doped nanoparticles using electric-pulsation. Taking a closer look at the uptake direction of exogenous materials will help researchers to understand an unknown uptake phenomenon in which way foreign materials are inclined to move, and furthermore to design functional nanoparticles for electroporative gene delivery.This paper presents direct visualization of uptake directionality for cell-membrane impermeant fluorescent molecules and fluorescence-doped nanoparticles at a single-cell level during electroporation. To observe directly the uptake direction, we used microchannel-type electroporation that can generate a relatively symmetric and uniform electric field. For all the image frames during electroporation, fluorescence intensities

Multi-spectral endogenous fluorescence imaging for bacterial differentiation

Science.gov (United States)

Chernomyrdin, Nikita V.; Babayants, Margarita V.; Korotkov, Oleg V.; Kudrin, Konstantin G.; Rimskaya, Elena N.; Shikunova, Irina A.; Kurlov, Vladimir N.; Cherkasova, Olga P.; Komandin, Gennady A.; Reshetov, Igor V.; Zaytsev, Kirill I.

2017-07-01

In this paper, the multi-spectral endogenous fluorescence imaging was implemented for bacterial differentiation. The fluorescence imaging was performed using a digital camera equipped with a set of visual bandpass filters. Narrowband 365 nm ultraviolet radiation passed through a beam homogenizer was used to excite the sample fluorescence. In order to increase a signal-to-noise ratio and suppress a non-fluorescence background in images, the intensity of the UV excitation was modulated using a mechanical chopper. The principal components were introduced for differentiating the samples of bacteria based on the multi-spectral endogenous fluorescence images.

MCNP calculation for calibration curve of X-ray fluorescence analysis

International Nuclear Information System (INIS)

Tan Chunming; Wu Zhifang; Guo Xiaojing; Xing Guilai; Wang Zhentao

2011-01-01

Due to the compositional variation of the sample, linear relationship between the element concentration and fluorescent intensity will not be well maintained in most X-ray fluorescence analysis. To overcome this, we use MCNP program to simulate fluorescent intensity of Fe (0∼100% concentration range) within binary mixture of Cr and O which represent typical strong absorption and weak absorption conditions respectively. The theoretic calculation shows that the relationship can be described as a curve determined by parameter p and value of p can be obtained with given absorption coefficient of substrate elements and element under detection. MCNP simulation results are consistent with theoretic calculation. Our research reveals that MCNP program can calculate the Calibration Curve of X-ray fluorescence very well. (authors)

Three gene expression vector sets for concurrently expressing multiple genes in Saccharomyces cerevisiae.

Science.gov (United States)

Ishii, Jun; Kondo, Takashi; Makino, Harumi; Ogura, Akira; Matsuda, Fumio; Kondo, Akihiko

2014-05-01

Yeast has the potential to be used in bulk-scale fermentative production of fuels and chemicals due to its tolerance for low pH and robustness for autolysis. However, expression of multiple external genes in one host yeast strain is considerably labor-intensive due to the lack of polycistronic transcription. To promote the metabolic engineering of yeast, we generated systematic and convenient genetic engineering tools to express multiple genes in Saccharomyces cerevisiae. We constructed a series of multi-copy and integration vector sets for concurrently expressing two or three genes in S. cerevisiae by embedding three classical promoters. The comparative expression capabilities of the constructed vectors were monitored with green fluorescent protein, and the concurrent expression of genes was monitored with three different fluorescent proteins. Our multiple gene expression tool will be helpful to the advanced construction of genetically engineered yeast strains in a variety of research fields other than metabolic engineering. © 2014 Federation of European Microbiological Societies. Published by John Wiley & Sons Ltd. All rights reserved.

Hybrid phosphorescence and fluorescence native spectroscopy for breast cancer detection.

Science.gov (United States)

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

2007-01-01

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

A Quantitative Fluorescence-Based Lipase Assay

Directory of Open Access Journals (Sweden)

Giovanna Lomolino

2012-01-01

Full Text Available An easy and fast gel diffusion assay for detecting and monitoring lipase activity by quantification of fluorescein is described. By measuring the intensity of fluorescein, it is possible to obtain a calibration curve with a regression coefficient better than by using the radius of fluorescent haloes. Through the quantification of fluorescence intensity of fluorescein released after the hydrolysis of a fluorescent ester, fluorescein dibutyrate, used as substrate in agar plates, commercial and skimmed milk lipase activity were studied. Moreover, with this method, lipase activity can be monitored in reaction medium that contains compounds which are affected by turbidity or cause measurement interference for UV-spectrophotometer and fluorimeter. In this experiment, boiled skimmed milk was dispersed in the agar gel with fluorescein dibutyrate, and it was used as a reaction medium to mimic natural conditions. The development of such an assay has a potential for applications in industries ranging from pharmaceuticals to food production and monitoring.

Multispectral open-air intraoperative fluorescence imaging.

Science.gov (United States)

Behrooz, Ali; Waterman, Peter; Vasquez, Kristine O; Meganck, Jeff; Peterson, Jeffrey D; Faqir, Ilias; Kempner, Joshua

2017-08-01

Intraoperative fluorescence imaging informs decisions regarding surgical margins by detecting and localizing signals from fluorescent reporters, labeling targets such as malignant tissues. This guidance reduces the likelihood of undetected malignant tissue remaining after resection, eliminating the need for additional treatment or surgery. The primary challenges in performing open-air intraoperative fluorescence imaging come from the weak intensity of the fluorescence signal in the presence of strong surgical and ambient illumination, and the auto-fluorescence of non-target components, such as tissue, especially in the visible spectral window (400-650 nm). In this work, a multispectral open-air fluorescence imaging system is presented for translational image-guided intraoperative applications, which overcomes these challenges. The system is capable of imaging weak fluorescence signals with nanomolar sensitivity in the presence of surgical illumination. This is done using synchronized fluorescence excitation and image acquisition with real-time background subtraction. Additionally, the system uses a liquid crystal tunable filter for acquisition of multispectral images that are used to spectrally unmix target fluorescence from non-target auto-fluorescence. Results are validated by preclinical studies on murine models and translational canine oncology models.

Inverse identification of intensity distributions from multiple flux maps in concentrating solar applications

International Nuclear Information System (INIS)

Erickson, Ben; Petrasch, Jörg

2012-01-01

Radiative flux measurements at the focal plane of solar concentrators are typically performed using digital cameras in conjunction with Lambertian targets. To accurately predict flux distributions on arbitrary receiver geometries directional information about the radiation is required. Currently, the directional characteristics of solar concentrating systems are predicted via ray tracing simulations. No direct experimental technique to determine intensities of concentrating solar systems is available. In the current paper, multiple parallel flux measurements at varying distances from the focal plane together with a linear inverse method and Tikhonov regularization are used to identify the directional and spatial intensity distribution at the solution plane. The directional binning feature of an in-house Monte Carlo ray tracing program is used to provide a reference solution. The method has been successfully applied to two-dimensional concentrators, namely parabolic troughs and elliptical troughs using forward Monte Carlo ray tracing simulations that provide the flux maps as well as consistent, associated intensity distribution for validation. In the two-dimensional case, intensity distributions obtained from the inverse method approach the Monte Carlo forward solution. In contrast, the method has not been successful for three dimensional and circular symmetric concentrator geometries.

Alteration of UV primary fluorescence of vital tumor cells following irradiation with neutrons and gamma rays

International Nuclear Information System (INIS)

Merkle, K.

1980-01-01

The change of UV primary fluorescence intensity of vital unstained cells of Ehrlich ascites carcinoma after 60 Co-gamma and neutron irradiation was investigated. The mean neutron energy was 6.2 MeV. Fluorescence intensity was detected using impulse cytophotometry. The UV intensity of single cells was measured in the spectral range from 300-400 nm. An monotonous increase of dose-effect curves and a maximum at 3.5 Gy (neutrons) and 30 Gy (γ-rays) was obtained. The first relevant increase of fluorescence intensity was detected at 0.4 Gy (neutrons) and 0.75 Gy (γ-rays). Factors influencing the increase and decrease of primary fluorescence behavior of vital cells are discussed. (author)

Anthracene Fluorescence Quenching by a Tetrakis (Ketocarboxamide Cavitand

Directory of Open Access Journals (Sweden)

Tibor Zoltan Janosi

2014-01-01

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

Cryo-imaging of fluorescently labeled single cells in a mouse

Science.gov (United States)

Steyer, Grant J.; Roy, Debashish; Salvado, Olivier; Stone, Meredith E.; Wilson, David L.

2009-02-01

We developed a cryo-imaging system to provide single-cell detection of fluorescently labeled cells in mouse, with particular applicability to stem cells and metastatic cancer. The Case cryoimaging system consists of a fluorescence microscope, robotic imaging positioner, customized cryostat, PC-based control system, and visualization/analysis software. The system alternates between sectioning (10-40 μm) and imaging, collecting color brightfield and fluorescent blockface image volumes >60GB. In mouse experiments, we imaged quantum-dot labeled stem cells, GFP-labeled cancer and stem cells, and cell-size fluorescent microspheres. To remove subsurface fluorescence, we used a simplified model of light-tissue interaction whereby the next image was scaled, blurred, and subtracted from the current image. We estimated scaling and blurring parameters by minimizing entropy of subtracted images. Tissue specific attenuation parameters were found [uT : heart (267 +/- 47.6 μm), liver (218 +/- 27.1 μm), brain (161 +/- 27.4 μm)] to be within the range of estimates in the literature. "Next image" processing removed subsurface fluorescence equally well across multiple tissues (brain, kidney, liver, adipose tissue, etc.), and analysis of 200 microsphere images in the brain gave 97+/-2% reduction of subsurface fluorescence. Fluorescent signals were determined to arise from single cells based upon geometric and integrated intensity measurements. Next image processing greatly improved axial resolution, enabled high quality 3D volume renderings, and improved enumeration of single cells with connected component analysis by up to 24%. Analysis of image volumes identified metastatic cancer sites, found homing of stem cells to injury sites, and showed microsphere distribution correlated with blood flow patterns. We developed and evaluated cryo-imaging to provide single-cell detection of fluorescently labeled cells in mouse. Our cryo-imaging system provides extreme (>60GB), micron

Laser induced fluorescence of dental caries

Science.gov (United States)

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

1988-01-01

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

Contribution of glue layer into epidermis sample fluorescence dynamics

Science.gov (United States)

Salomatina, Elena V.; Chernova, Svetlana P.; Pravdin, Alexander B.

2000-04-01

In this work, the temporal behavior of autofluorescence of epidermis samples under UV-irradiation has ben studied. The samples were prepared using surface epidermis stripping technique. Fluorescence spectra and kinetic curves of fluorescence intensity have been obtained. It has been concluded that the glue composition used allows the measurement of epidermis fluorescence dynamics with the first 60 min of experiment.

An off-on Fluorescent Sensor for Detecting a Wide Range of Water Content in Organic Solvents

Energy Technology Data Exchange (ETDEWEB)

Kim, Kanghyeon; Lee, Wanjin; Kim, Jae Nyoung; Kim, Hyung Jin [Chonnam National Univ., Gwangju (Korea, Republic of)

2013-08-15

This paper describes the synthesis and water sensing properties of a fluorescent photoinduced electron transfer (PET) sensor (5) with an extended operating sensing range. The 1,8-naphthalimide derivative (5) attached with a piperazine group and a carboxylic group was synthesized and applied as a fluorescent water sensor in water-miscible organic solvents. The fluorescence intensity of the dye 5 increased with increasing water content up to 80% (v/v) and the fluorescence intensities were enhanced 45-, 67- and 122-fold in aqueous EtOH, DMF and DMSO solutions, respectively. In aqueous acetone solution, the enhancement of the fluorescence intensities was somewhat lower (30-fold) but the response range was wider (0-90%, v/v)

A novel fluorescent retrograde neural tracer: cholera toxin B conjugated carbon dots

Science.gov (United States)

Zhou, Nan; Hao, Zeyu; Zhao, Xiaohuan; Maharjan, Suraj; Zhu, Shoujun; Song, Yubin; Yang, Bai; Lu, Laijin

2015-09-01

The retrograde neuroanatomical tracing method is a key technique to study the complex interconnections of the nervous system. Traditional tracers have several drawbacks, including time-consuming immunohistochemical or immunofluorescent staining procedures, rapid fluorescence quenching and low fluorescence intensity. Carbon dots (CDs) have been widely used as a fluorescent bio-probe due to their ultrasmall size, excellent optical properties, chemical stability, biocompatibility and low toxicity. Herein, we develop a novel fluorescent neural tracer: cholera toxin B-carbon dot conjugates (CTB-CDs). It can be taken up and retrogradely transported by neurons in the peripheral nervous system of rats. Our results show that CTB-CDs possess high photoluminescence intensity, good optical stability, a long shelf-life and non-toxicity. Tracing with CTB-CDs is a direct and more economical way of performing retrograde labelling experiments. Therefore, CTB-CDs are reliable fluorescent retrograde tracers.The retrograde neuroanatomical tracing method is a key technique to study the complex interconnections of the nervous system. Traditional tracers have several drawbacks, including time-consuming immunohistochemical or immunofluorescent staining procedures, rapid fluorescence quenching and low fluorescence intensity. Carbon dots (CDs) have been widely used as a fluorescent bio-probe due to their ultrasmall size, excellent optical properties, chemical stability, biocompatibility and low toxicity. Herein, we develop a novel fluorescent neural tracer: cholera toxin B-carbon dot conjugates (CTB-CDs). It can be taken up and retrogradely transported by neurons in the peripheral nervous system of rats. Our results show that CTB-CDs possess high photoluminescence intensity, good optical stability, a long shelf-life and non-toxicity. Tracing with CTB-CDs is a direct and more economical way of performing retrograde labelling experiments. Therefore, CTB-CDs are reliable fluorescent retrograde

Fluorescence spectral studies of Gum Arabic: Multi-emission of Gum Arabic in aqueous solution

Energy Technology Data Exchange (ETDEWEB)

Dhenadhayalan, Namasivayam, E-mail: ndhena@gmail.com [Department of Chemistry, National Taiwan University, Taipei, Taiwan (China); Mythily, Rajan, E-mail: rajanmythily@gmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600 106 (India); Kumaran, Rajendran, E-mail: kumaranwau@rediffmail.com [Department of Chemistry, Dwaraka Doss Goverdhan Doss Vaishnav College (Autonomous), 833, Gokul Bagh, E.V.R. Periyar Road, Arumbakkam, Chennai 600 106 (India)

2014-11-15

Gum Arabic (GA), a food hydrocolloid is a natural composite obtained from the stems and branches of Acacia Senegal and Acacia Seyal trees. GA structure is made up of highly branched arabinogalactan polysaccharides. Steady-state absorption, fluorescence, and time-resolved fluorescence spectral studies of acid hydrolyzed GA solutions were carried out at various pH conditions. The fluorescence in GA is predominantly attributed to the presence of tyrosine and phenylalanine amino acids. The presence of multi-emissive peaks at different pH condition is attributed to the exposure of the fluorescing amino acids to the aqueous phase, which contains several sugar units, hydrophilic and hydrophobic moieties. Time-resolved fluorescence studies of GA exhibits a multi-exponential decay with different fluorescence lifetime of varying amplitude which confirms that tyrosine is confined to a heterogeneous microenvironment. The existence of multi-emissive peaks with large variation in the fluorescence intensities were established by 3D emission contour spectral studies. The probable location of the fluorophore in a heterogeneous environment was further ascertained by constructing a time-resolved emission spectrum (TRES) and time-resolved area normalized emission spectrum (TRANES) plots. Fluorescence spectral technique is used as an analytical tool in understanding the photophysical properties of a water soluble complex food hydrocolloid containing an intrinsic fluorophore located in a multiple environment is illustrated. - Highlights: • The Manuscript deals with the steady state absorption, emission, fluorescence lifetime and time-resolved emission spectrum studies of Gum Arabic in aqueous medium at various pH conditions. • The fluorescence emanates from the tyrosine amino acid present in GA. • Change in pH results in marked variation in the fluorescence spectral properties of tyrosine. • Fluorescence spectral techniques are employed as a tool in establishing the

Detection of vegetation stress from laser-induced fluorescence signatures

International Nuclear Information System (INIS)

Subhash, N.

1995-01-01

The in vivo laser-induced fluorescence (LIF) signatures of UV irradiated Salvia splendens plants were measured using an Optical Multichannel Analyser (OMA) system with Nitrogen laser excitation. The LIF spectra which consisted of the blue-green and the red chlorophyll bands were analysed with a non-linear interactive procedure using Gaussian spectral functions. The fluorescence intensity ratios of the various bands obtained from curve fitted parameters were found to be more sensitive to changes in the photosynthetic activity of the plant. The variation in the intensity ratio for the chlorophyll bands for nutrient stressed sunflower, cotton and groundnut plants as well as the nutrient and water stressed rice plants are also presented. It is observed that vegetation stress not only changes the fluorescence intensity ratios and the vitality index of the plant but also changes the peak position of the emission bands, in some cases. It is also seen that analysis of the fluorescence spectra in vegetation remote sensing applications would require a deconvolution procedure to evaluate the exact contribution of each band in the total spectra. (author). 23 refs, 8 figs, 5 tabs

Detecting crop population growth using chlorophyll fluorescence imaging.

Science.gov (United States)

Wang, Heng; Qian, Xiangjie; Zhang, Lan; Xu, Sailong; Li, Haifeng; Xia, Xiaojian; Dai, Liankui; Xu, Liang; Yu, Jingquan; Liu, Xu

2017-12-10

For both field and greenhouse crops, it is challenging to evaluate their growth information on a large area over a long time. In this work, we developed a chlorophyll fluorescence imaging-based system for crop population growth information detection. Modular design was used to make the system provide high-intensity uniform illumination. This system can perform modulated chlorophyll fluorescence induction kinetics measurement and chlorophyll fluorescence parameter imaging over a large area of up to 45  cm×34  cm. The system can provide different lighting intensity by modulating the duty cycle of its control signal. Results of continuous monitoring of cucumbers in nitrogen deficiency show the system can reduce the judge error of crop physiological status and improve monitoring efficiency. Meanwhile, the system is promising in high throughput application scenarios.

Conformational changes induced by Mg2+ on the multiple forms of glutamine synthetase from Bacillus brevis Bb G1

Directory of Open Access Journals (Sweden)

Suja Abraham

2013-01-01

Full Text Available Conformational changes play an important role in the function of proteins. Glutamine synthetase, an important enzyme of nitrogen metabolism, was purified under sporulating (GSala and non-sporulating (GSpyr conditions and the effect of Mg2+ on these multiple forms was studied by fluorescence spectroscopy to detect possible conformational changes that occur in the presenceof Mg2+. The substantial changes in the fluorescence emission maximum, fluorescence intensity and lifetime that occur in the presence of different concentrations of Mg2+, indicated major changes in molecular conformations in both forms of this enzyme. The fluorescent changes produced by the effect of Mg2+ in GSala was much more prominent than in GSpyr. These observations strongly support the possibility that GSala and GSpyr undergoes a conformational change on binding with Mg2+.

Ratiometric fluorescent nanoparticles for sensing temperature

Energy Technology Data Exchange (ETDEWEB)

Peng, Hong-Shang, E-mail: hillphs@yahoo.com.cn; Huang, Shi-Hua [Beijing Jiaotong University, Key Laboratory of Luminescence and Optical Information, Ministry of Education, Institute of Optoelectronic Technology (China); Wolfbeis, Otto S. [University of Regensburg, Institute of Analytical Chemistry, Chemo- and Biosensors (Germany)

2010-10-15

A ratiometric type of fluorescent nanoparticle was prepared via an encapsulation-reprecipitation method. By introducing an alkoxysilanized dye as a reference, the nanoparticles (NPs) give both a green and a red fluorescence under one single-wavelength excitation. The resulted ratiometric fluorescence is found to be highly temperature-dependent in the physiological range (25-45 {sup o}C), with an intensity temperature sensitivity of -4.0%/{sup o}C. Given the small size (20-30 nm in diameter) and biocompatible nature (silica out layer), such kind of NPs were very promising as temperature nanosensors for cellular sensing and imaging.

Experimental station for gas phase fluorescence spectroscopy

International Nuclear Information System (INIS)

Stankiewicz, M.; Garcia, E. Melero; Ruiz, J. Alvarez; Erman, P.; Hatherly, P.A.; Kivimaeki, A.; Rachlew, E.; Rius i Riu, J.

2004-01-01

The details of an experimental setup for gas phase atomic and molecular fluorescence measurements using synchrotron radiation are described in this article. The most significant part of the apparatus is an optical arrangement, which allows for simultaneous measurements of dispersed as well as total fluorescence intensity using an effusive gas jet and an inbuilt gas cell assembled in a convenient plug and measure configuration. The first measurements concerning fluorescence of the N 2 molecule around the N 1s edge obtained with this setup are presented

Fluorescence enhancement of CdTe/CdS quantum dots by coupling of glyphosate and its application for sensitive detection of copper ion

Energy Technology Data Exchange (ETDEWEB)

Liu Zhengqing; Liu Shaopu; Yin Pengfei [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China); He Youqiu, E-mail: heyq@swu.edu.cn [Key Laboratory on Luminescence and Real-Time Analysis, Ministry of Education, School of Chemistry and Chemical Engineering, Southwest University, Chongqing 400715 (China)

2012-10-01

Graphical abstract: Glyphosate (Glyp) had been used to modify the surface of CdTe/CdS QDs, resulting in the enhancement of fluorescence intensity. The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu{sup 2+} based on the fluorescence quenching. Highlights: Black-Right-Pointing-Pointer Water soluble CdTe/CdS quantum dots capped with glyphosate were firstly synthesized. Black-Right-Pointing-Pointer The fluorescence of the Glyp-functionalized QDs was quenched by copper ion. Black-Right-Pointing-Pointer A new fluorescent sensor for copper ion was developed based on the prepared QDs. Black-Right-Pointing-Pointer The sensor exhibited high sensitivity and good selectivity for copper ion. - Abstract: A novel fluorescent probe for Cu{sup 2+} determination based on the fluorescence quenching of glyphosate (Glyp)-functionalized quantum dots (QDs) was firstly reported. Glyp had been used to modify the surface of QDs to form Glyp-functionalized QDs following the capping of thioglycolic acid on the core-shell CdTe/CdS QDs. Under the optimal conditions, the response was linearly proportional to the concentration of Cu{sup 2+} between 2.4 Multiplication-Sign 10{sup -2} {mu}g mL{sup -1} and 28 {mu}g mL{sup -1}, with a detection limit of 1.3 Multiplication-Sign 10{sup -3} {mu}g mL{sup -1} (3{delta}). The Glyp-functionalized QDs fluorescent probe offers good sensitivity and selectivity for detecting Cu{sup 2+}. The fluorescent probe was successfully used for the determination of Cu{sup 2+} in environmental samples. The mechanism of reaction was also discussed.

Laser induced fluorescence of some plant leaves

International Nuclear Information System (INIS)

Helmi, M.S.; Mohamed, M.M.; Amer, R.; Elshazly, O.; Elraey, M.

1992-01-01

Laser induced fluorescence (LIF) is successfully used as a technique for remote detection of spectral characteristics of some plants. A pulsed nitrogen laser at 337.1 nm is used to excite cotton, corn and rice leaves. The fluorescence spectrum is detected in the range from 340 nm to 820 nm. It is found that, these plant leaves have common fluorescence maxima at 440 nm, 685 nm and 740 nm. plant leaves are also found to be identifiable by the ratio of the fluorescence intensity at 440 nm to that at 685 nm. The present technique can be further used as a means of assessing, remotely, plant stresses. 5 fig

Laser-Induced Fluorescence (LIF) from plant foliage

Science.gov (United States)

Chappelle, Emmett W.; Williams, Darrel L.

1987-01-01

The fluorescence spectra and fluorescence induction kinetics of green plants excited at 337 nm by a laser were studied. They correlate with plant type, as well as with changes in the physiology of the plant as the result of stress. The plant types studied include herbaceous dicots, monocots, hardwoods, conifers, and algae. These plant types could be identified on the basis of differences in either the number of fluorescent bands or the relative intensity of the bands. Differences in fluorescent spectra which could be related to vigor status are observed in conifers located in an area of high atmospheric deposition. Changes in the fluorescence spectra and induction kinetics are also seen in plants grown under conditions of nutrient deficiency and drought stress.

Thermal maturity of Tasmanites microfossils from confocal laser scanning fluorescence microscopy

Science.gov (United States)

Hackley, Paul C.; Kus, Jolanta

2015-01-01

We report here, for the first time, spectral properties of Tasmanites microfossils determined by confocal laser scanning fluorescence microscopy (CLSM, using Ar 458 nm excitation). The Tasmanites occur in a well-characterized natural maturation sequence (Ro 0.48–0.74%) of Devonian shale (n = 3 samples) from the Appalachian Basin. Spectral property λmax shows excellent agreement (r2 = 0.99) with extant spectra from interlaboratory studies which used conventional fluorescence microscopy techniques. This result suggests spectral measurements from CLSM can be used to infer thermal maturity of fluorescent organic materials in geologic samples. Spectra of regions with high fluorescence intensity at fold apices and flanks in individual Tasmanites are blue-shifted relative to less-deformed areas in the same body that have lower fluorescence intensity. This is interpreted to result from decreased quenching moiety concentration at these locations, and indicates caution is needed in the selection of measurement regions in conventional fluorescence microscopy, where it is common practice to select high intensity regions for improved signal intensity and better signal to noise ratios. This study also documents application of CLSM to microstructural characterization of Tasmanites microfossils. Finally, based on an extant empirical relation between conventional λmax values and bitumen reflectance, λmax values from CLSM of Tasmanites microfossils can be used to calculate a bitumen reflectance equivalent value. The results presented herein can be used as a basis to broaden the future application of CLSM in the geological sciences into hydrocarbon prospecting and basin analysis.

Graphene oxide-sensitized molecularly imprinted opto-polymers for charge-transfer fluorescent sensing of cyanoguanidine.

Science.gov (United States)

Liu, Huilin; Zhou, Kaiwen; Chen, Xiaomo; Wang, Jing; Wang, Shuo; Sun, Baoguo

2017-11-15

The hierarchical structuring of materials offers exciting opportunities to construct functional sensors. Multiple processes were combined to create complex materials for the selective detection of cyanoguanidine (CYA) using graphene oxide-sensitized molecularly imprinted opto-polymers (MIOP). Molecular imprinting was used to construct molecular-scale analyte-selective cavities, graphene oxide was introduced to provide a platform for the polymerization, and increase the stability and binding kinetic properties, and 3-methacryloxy propyl trimethoxy silane-modified quantum dots were combined with a functional monomer to increase the fluorescence quantum yield. Polymer cross-linking and fluorescence intensity were optimized for molecular recognition and opto-sensing detection. Selective and sensitive, fluorescence sensing of CYA was possible at concentrations as low as to 1.6μM. It could be applied to the rapid and cost-effective monitoring of CYA in infant formula. The approach is generic and applicable to many molecules and conventional opto-sensors, based on molecularly imprinted polymer formulations, individually or in multiplexed arrays. Copyright © 2017 Elsevier Ltd. All rights reserved.

Control of excitation in the fluorescence microscope.

Science.gov (United States)

Lea, D J; Ward, D J

1979-01-01

In fluorescence microscopy image brightness and contrast and the rate of fading depend upon the intensity of illumination of the specimen. An iris diaphragm or neutral density filters may be used to reduce fluorescence excitation. Also the excitation bandwidth may be varied by using a broad band exciter filter with a set of interchangeable yellow glass filters at the lamphouse.

Fluorescence excitation involving multiple electron transition states of N{sub 2} and CO{sub 2}

Energy Technology Data Exchange (ETDEWEB)

Wu, C.Y.R.; Chen, F.Z.; Hung, T.; Judge, D.L. [Univ. of Southern California, Los Angeles, CA (United States)

1997-04-01

The electronic states and electronic structures of N{sub 2} and CO{sub 2} in the 8-50 eV energy region have been studied extensively both experimentally and theoretically. In the energy region higher than 25 eV there exists many electronic states including multiple electron transition (MET) states which are responsible for producing most of the dissociative photoionization products. The electronic states at energies higher than 50 eV have been mainly determined by Auger spectroscopy, double charge transfer, photofragment spectroscopy and ion-ion coincidence spectroscopy. The absorption and ionization spectra of these molecules at energies higher than 50 eV mainly show a monotonic decrease in cross section values and exhibit structureless features. The decay channels of MET and Rydberg (or superexcited) states include autoionization, ionization, dissociative ionization, predissociation, and dissociation while those of single ion and multiple ion states may involve predissociation. and dissociation processes. The study of fluorescence specifically probes electronically excited species resulting from the above-mentioned decay channels and provides information for understanding the competition among these channels.

Development of ultraviolet LED devices containing europium (III) complexes in fluorescence layer

International Nuclear Information System (INIS)

Iwanaga, Hiroki; Amano, Akio; Aiga, Fumihiko; Harada, Kohichi; Oguchi, Masayuki

2006-01-01

Relations between molecular structures of europium complexes and their luminescent properties were investigated. Europium complex with β-diketones and two different phosphine oxides 8 was highly soluble in fluorinated medium, and realized largest fluorescence intensities. The luminous intensity of ultraviolet light emitting diodes devices (LEDs) whose fluorescence layer consists of fluorinated polymer and 8 was over 200 mcd (20 mA). Fluorescence compounds of this type are promising for application in next-generation white LEDs. Moreover, we proposed a novel molecular design of europium complex with asymmetric diphosphine dioxide

Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa.

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Schneidereit, D; Vass, H; Reischl, B; Allen, R J; Friedrich, O

2016-01-01

The fluorescent Ca2+ sensitive dyes Fura Red (ratiometric) and Fluo-4 (non-ratiometric) are widely utilized for the optical assessment of Ca2+ fluctuations in vitro as well as in situ. The fluorescent behavior of these dyes is strongly depends on temperature, pH, ionic strength and pressure. It is crucial to understand the response of these dyes to pressure when applying calcium imaging technologies in the field of high pressure bioscience. Therefore, we use an optically accessible pressure vessel to pressurize physiological Ca2+-buffered solutions at different fixed concentrations of free Ca2+ (1 nM to 25.6 μM) and a specified dye concentration (12 μM) to pressures of 200 MPa, and record dye fluorescence intensity. Our results show that Fluo-4 fluorescence intensity is reduced by 31% per 100 MPa, the intensity of Fura Red is reduced by 10% per 100 MPa. The mean reaction volume for the dissociation of calcium from the dye molecules [Formula: see text] is determined to -17.8 ml mol-1 for Fluo-4 and -21.3 ml mol-1 for Fura Red. Additionally, a model is presented that is used to correct for pressure-dependent changes in pH and binding affinity of Ca2+ to EGTA, as well as to determine the influence of these changes on dye fluorescence.

Calcium Sensitive Fluorescent Dyes Fluo-4 and Fura Red under Pressure: Behaviour of Fluorescence and Buffer Properties under Hydrostatic Pressures up to 200 MPa

Science.gov (United States)

Vass, H.; Reischl, B.; Allen, R. J.; Friedrich, O.

2016-01-01

The fluorescent Ca2+ sensitive dyes Fura Red (ratiometric) and Fluo-4 (non-ratiometric) are widely utilized for the optical assessment of Ca2+ fluctuations in vitro as well as in situ. The fluorescent behavior of these dyes is strongly depends on temperature, pH, ionic strength and pressure. It is crucial to understand the response of these dyes to pressure when applying calcium imaging technologies in the field of high pressure bioscience. Therefore, we use an optically accessible pressure vessel to pressurize physiological Ca2+-buffered solutions at different fixed concentrations of free Ca2+ (1 nM to 25.6 μM) and a specified dye concentration (12 μM) to pressures of 200 MPa, and record dye fluorescence intensity. Our results show that Fluo-4 fluorescence intensity is reduced by 31% per 100 MPa, the intensity of Fura Red is reduced by 10% per 100 MPa. The mean reaction volume for the dissociation of calcium from the dye molecules Δdv¯ is determined to -17.8 ml mol-1 for Fluo-4 and -21.3 ml mol-1 for Fura Red. Additionally, a model is presented that is used to correct for pressure-dependent changes in pH and binding affinity of Ca2+ to EGTA, as well as to determine the influence of these changes on dye fluorescence. PMID:27764134

Multiplex and high-throughput DNA detection using surface plasmon mediated fluorescence

Science.gov (United States)

Mei, Zhong

The overall objective of this research project was to develop a user-friendly and sensitive biosensor for nucleic acid aptamers with multiplexing and high-throughput capability. The sensing was based on the fluorescence signals emitted by the fluorophores coupling with plamonic nanoparticle (gold nanorod) deposited on a patterned substrate. Gold nanorods (GNRs) were synthesized using a binary mixture of hexadecyltrimethylammonium bromide (CTAB) and sodium oleate (NaOL) in seed mediated growth method. Polytetrafluoroethylene (PTFE) printed glass slides were selectively coated with a gold thin-film to define hydrophilic areas for GNR deposition. Due to the wettablity contrast, GNR solution dropped on the slide was induced to assemble exclusively in the hydrophilic spots. By controlling temperature and humidity of the evaporation process, vertically-standing GNR arrays were achieved on the pattered slide. Fluorescence was conjugated to GNR surface via DNA double strand with tunable length. Theoretical simulation predicted a flat layer ( 30 nm thick) of uniform "hot spots" presented on the GNR tips, which could modify the nearby fluorescence. Experimentally, the vertical GNR arrays yielded metallic enhanced fluorescence (MEF) effect, which was dependent on the spectrum overlap and GNR-fluorophore distance. Specifically, the maximum enhancement of Quasar 670 and Alexa 750 was observed when it was coupled with GNR664 (plasmonic wavelength 664 nm) and GNR778 respectively at a distance of 16 nm, while the carboxyfluorescein (FAM) was at maximal intensity when attached to gold nanosphere520. This offers an opportunity for multiplexed DNA sensing. Based on this, we developed a novel GNR mediated fluorescence biosensor for DNA detection. Fluorescence labeled haipin-DNA probes were introduced to designated spots of GNR array with the matching LSPR wavelengths on the substrate. The fluorescence was quenched originally because of Forster resonance energy transfer (FRET) effect

Fluorescence from colours used for japanese painting under N_2 laser excitation

OpenAIRE

Miyoshi, Tadaki

1988-01-01

The fluorescence spectra have been measured for various colours used for Japanese painting in order to identify the pigments in paintings. Fluorescence was observed in various natural and synthetic colours. The fluorescence intensities of these colours are generally weaker than those of oil colours.

Quantification of Coffea arabica and Coffea canephora var. robusta concentration in blends by means of synchronous fluorescence and UV-Vis spectroscopies.

Science.gov (United States)

Dankowska, A; Domagała, A; Kowalewski, W

2017-09-01

The potential of fluorescence, UV-Vis spectroscopies as well as the low- and mid-level data fusion of both spectroscopies for the quantification of concentrations of roasted Coffea arabica and Coffea canephora var. robusta in coffee blends was investigated. Principal component analysis was used to reduce data multidimensionality. To calculate the level of undeclared addition, multiple linear regression (PCA-MLR) models were used with lowest root mean square error of calibration (RMSEC) of 3.6% and root mean square error of cross-validation (RMSECV) of 7.9%. LDA analysis was applied to fluorescence intensities and UV spectra of Coffea arabica, canephora samples, and their mixtures in order to examine classification ability. The best performance of PCA-LDA analysis was observed for data fusion of UV and fluorescence intensity measurements at wavelength interval of 60nm. LDA showed that data fusion can achieve over 96% of correct classifications (sensitivity) in the test set and 100% of correct classifications in the training set, with low-level data fusion. The corresponding results for individual spectroscopies ranged from 90% (UV-Vis spectroscopy) to 77% (synchronous fluorescence) in the test set, and from 93% to 97% in the training set. The results demonstrate that fluorescence, UV, and visible spectroscopies complement each other, giving a complementary effect for the quantification of roasted Coffea arabica and Coffea canephora var. robusta concentration in blends. Copyright © 2017 Elsevier B.V. All rights reserved.

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

International Nuclear Information System (INIS)

Ozawa, L.; Hersh, H.N.

1976-01-01

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

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

Science.gov (United States)

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

2017-09-01

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

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

Science.gov (United States)

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

1999-07-01

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

Tomato seeds maturity detection system based on chlorophyll fluorescence

Science.gov (United States)

Li, Cuiling; Wang, Xiu; Meng, Zhijun

2016-10-01

Chlorophyll fluorescence intensity can be used as seed maturity and quality evaluation indicator. Chlorophyll fluorescence intensity of seed coats is tested to judge the level of chlorophyll content in seeds, and further to judge the maturity and quality of seeds. This research developed a detection system of tomato seeds maturity based on chlorophyll fluorescence spectrum technology, the system included an excitation light source unit, a fluorescent signal acquisition unit and a data processing unit. The excitation light source unit consisted of two high power LEDs, two radiators and two constant current power supplies, and it was designed to excite chlorophyll fluorescence of tomato seeds. The fluorescent signal acquisition unit was made up of a fluorescence spectrometer, an optical fiber, an optical fiber scaffolds and a narrowband filter. The data processing unit mainly included a computer. Tomato fruits of green ripe stage, discoloration stage, firm ripe stage and full ripe stage were harvested, and their seeds were collected directly. In this research, the developed tomato seeds maturity testing system was used to collect fluorescence spectrums of tomato seeds of different maturities. Principal component analysis (PCA) method was utilized to reduce the dimension of spectral data and extract principal components, and PCA was combined with linear discriminant analysis (LDA) to establish discriminant model of tomato seeds maturity, the discriminant accuracy was greater than 90%. Research results show that using chlorophyll fluorescence spectrum technology is feasible for seeds maturity detection, and the developed tomato seeds maturity testing system has high detection accuracy.

A highly sensitive fluorescent probe based on BODIPY for Hg2+ in aqueous solution

Directory of Open Access Journals (Sweden)

ZHAO Junwei

2016-12-01

Full Text Available A highly sensitive fluorescent probe based on BODIPY and hydrazine for Hg2+ was designed and synthesized.This probe could detect mercury ions in aqueous solutions within 5 min.With the increase of Hg2+ mole concentration,an obvious red shift of UV-Vis absorption wavelength was observed and the fluorescence intensity significantly enhanced.It was found that the fluorescence intensity of an aqueous solution containing 0.1 μmol/L Hg2+ is much stronger than that of blank solution,which indicats that the fluorescent probe has high sensitivity.In addition,other metal ions could not cause the change of fluorescent spectra,which means this probe has good selectivity,as well.

Fluorescent properties of novel dendrimer dyes based on thiazole orange

International Nuclear Information System (INIS)

Fei Xuening; Gu Yingchun; Lan Yunquan; Shi Bin

2011-01-01

In this paper, polyamidoamine (PAMAM) dendrimers with active amino group of some generations (G=0.5-2) were prepared from commercial aminoacetaldehyde diethyl acetal by the divergent method. After that, thiazole orange (TO) with -COOH was incorporated with dendrimers of G=1 and 2 to afford novel dendrimer-TO dyes. The fluorescent properties studies showed that the fluorescent intensity of the same concentration of dendrimer-TO (G=2) was higher than that of the dendrimer-TO (G=1), and both of them were much stronger than free TO with -COOH. There was a fluorescent enhancement of the dendrimer dyes compared with free dye. The dendrimer dyes were of well-defined chemical structure,with little aggregation and self-quenching as well as good fluorescence properties of good stability, high intensity and sensitivity, which could be used in labeling cancer cells and further in diagnosis and detection of early-stage tumors. - Highlights: → A kind of dendrimer probe based on TO was designed and synthesized. → Dendrimers showed an obvious fluorescence enhancement compared to free dye. → Dendrimers labeled with BSA also showed fluorescence enhancement. → Dendrimers may be used in diagnosis and detection of early-stage tumors.

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

Science.gov (United States)

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.

Optical properties of flexible fluorescent films prepared by screen printing technology

Directory of Open Access Journals (Sweden)

Yan Chen

2018-05-01

Full Text Available 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(InN 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.

Optical properties of flexible fluorescent films prepared by screen printing technology

Science.gov (United States)

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.

Cooperative effects and slow dynamics of fluorescence intensity from quantum emitters in a dielectric

International Nuclear Information System (INIS)

Lozing, N.A.; Gladush, M.G.

2017-01-01

We study theoretically the possibility of spontaneous switching between dim and bright fluorescence modes from a cooperative ensemble of two-level atoms driven by a cw-laser. A numerical analysis of transient regimes and transformations of the fluorescence spectrum are reported.

Preliminary study on X-ray fluorescence computed tomography imaging of gold nanoparticles: Acceleration of data acquisition by multiple pinholes scheme

Science.gov (United States)

Sasaya, Tenta; Sunaguchi, Naoki; Seo, Seung-Jum; Hyodo, Kazuyuki; Zeniya, Tsutomu; Kim, Jong-Ki; Yuasa, Tetsuya

2018-04-01

Gold nanoparticles (GNPs) have recently attracted attention in nanomedicine as novel contrast agents for cancer imaging. A decisive tomographic imaging technique has not yet been established to depict the 3-D distribution of GNPs in an object. An imaging technique known as pinhole-based X-ray fluorescence computed tomography (XFCT) is a promising method that can be used to reconstruct the distribution of GNPs from the X-ray fluorescence emitted by GNPs. We address the acceleration of data acquisition in pinhole-based XFCT for preclinical use using a multiple pinhole scheme. In this scheme, multiple projections are simultaneously acquired through a multi-pinhole collimator with a 2-D detector and full-field volumetric beam to enhance the signal-to-noise ratio of the projections; this enables fast data acquisition. To demonstrate the efficacy of this method, we performed an imaging experiment using a physical phantom with an actual multi-pinhole XFCT system that was constructed using the beamline AR-NE7A at KEK. The preliminary study showed that the multi-pinhole XFCT achieved a data acquisition time of 20 min at a theoretical detection limit of approximately 0.1 Au mg/ml and at a spatial resolution of 0.4 mm.

Detection of rheumatoid arthritis in humans by fluorescence imaging

Science.gov (United States)

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

2010-02-01

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

Nanostructures Derived from Starch and Chitosan for Fluorescence Bio-Imaging

Science.gov (United States)

Zu, Yinxue; Bi, Jingran; Yan, Huiping; Wang, Haitao; Song, Yukun; Zhu, Bei-Wei; Tan, Mingqian

2016-01-01

Fluorescent nanostructures (NSs) derived from polysaccharides have drawn great attention as novel fluorescent probes for potential bio-imaging applications. Herein, we reported a facile alkali-assisted hydrothermal method to fabricate polysaccharide NSs using starch and chitosan as raw materials. Transmission electron microscopy (TEM) demonstrated that the average particle sizes are 14 nm and 75 nm for starch and chitosan NSs, respectively. Fourier transform infrared (FT-IR) spectroscopy analysis showed that there are a large number of hydroxyl or amino groups on the surface of these polysaccharide-based NSs. Strong fluorescence with an excitation-dependent emission behaviour was observed under ultraviolet excitation. Interestingly, the photostability of the NSs was found to be superior to fluorescein and rhodamine B. The quantum yield of starch NSs could reach 11.12% under the excitation of 360 nm. The oxidative metal ions including Cu(II), Hg(II)and Fe(III) exhibited a quench effect on the fluorescence intensity of the prepared NSs. Both of the two kinds of the multicoloured NSs showed a maximum fluorescence intensity at pH 7, while the fluorescence intensity decreased dramatically when they were put in an either acidic or basic environment (at pH 3 or 11). The cytotoxicity study of starch NSs showed that low cell cytotoxicity and 80% viability was found after 24 h incubation, when their concentration was less than 10 mg/mL. The study also showed the possibility of using the multicoloured starch NSs for mouse melanoma cells and guppy fish imaging. PMID:28335258

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

Directory of Open Access Journals (Sweden)

Jieni Yao

2018-05-01

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

Fluorescence spectral properties of stomach tissues with pathology

Science.gov (United States)

Giraev, K. M.; Ashurbekov, N. A.; Lahina, M. A.

2012-05-01

Steady-state fluorescence and diffuse reflection spectra are measured for in vivo normal and pathological (chronic atrophic and ulcerating defects, malignant neoplasms) stomach mucous lining tissues. The degree of distortion of the fluorescence spectra is estimated taking light scattering and absorption into account. A combination of Gauss and Lorentz functions is used to decompose the fluorescence spectra. Potential groups of fluorophores are determined and indices are introduced to characterize the dynamics of their contributions to the resultant spectra as pathologies develop. Reabsorption is found to quench the fluorescence of structural proteins by as much as a factor of 3, while scattering of the light can increase the fluorescence intensity of flavin and prophyrin groups by as much as a factor of 2.

Fluorescence ON–OFF switching using micelle of stimuli-responsive double hydrophilic block copolymers: Nile Red fluorescence in micelles of poly(acrylic acid-b-N-isopropylacrylamide)

Energy Technology Data Exchange (ETDEWEB)

Yee, Min Min; Tsubone, Miyabi; Morita, Takuya [Department of Chemistry, Graduate School of Science & Engineering, Saga University, 1 Honjo, Saga 840-8502 (Japan); Yusa, Shin-ichi [Department of Materials Science and Chemistry, University of Hyogo, 2167 Shosha, Himeji 671-2280 (Japan); Nakashima, Kenichi, E-mail: nakashik@cc.saga-u.ac.jp [Department of Chemistry, Graduate School of Science & Engineering, Saga University, 1 Honjo, Saga 840-8502 (Japan)

2016-08-15

The dual-mode fluorescence ON–OFF switching of Nile Red (NR) by using stimuli-responsive polymeric micelle of poly(acrylic acid-b-N-isopropylacrylamide) (PAA-b-PNIPAM) has been studied. PAA-b-PNIPAM, one of double hydrophilic block copolymers, is known to form PNIPAM-core/PAA-corona micelles in aqueous solutions when the temperature of the solution is elevated up to the lower critical solution temperature (LCST) of PNIPAM block. It also forms PAA-core/PNIPAM-corona micelles when the anionic PAA block is charge-neutralized with cationic cetyltrimethylammonium ion. Fluorescence properties of NR in the micelles are elucidated by observing various fluorescence parameters such as intensity, polarization, and quantum yield. It is found that the fluorescence intensity is negligibly low (OFF-state) when PAA-b-PNIPAM exists as a form of unimer, whereas it is remarkably enhanced (ON-state) when the PNIPAM-core or PAA-core micelles are formed. These results demonstrate that a novel fluorescence ON–OFF switching system can be constructed by using PAA-b-PNIPAM micelles and NR.

A study of fluorescence properties of citrinin in β-cyclodextrin aqueous solution and different solvents

International Nuclear Information System (INIS)

Zhou Youxiang; Chen Jianbiao; Dong Lina; Lu Liang; Chen Fusheng; Hu Dingjin; Wang Xiaohong

2012-01-01

Citrinin (CIT) is a nephrotoxic mycotoxin initially isolated from filamentous fungus Penicilliu citrinum. It was later isolated from several other species, such as Aspergillus and Monascus. It has a conjugated, planar structure that gives it a natural fluorescence ability, which can be used to develop sensitive methods for detecting CIT in food. In this paper, we used the spectrofluorescence technique to study the effects of pH value, β-cyclodextrin (β-CD) and organic solvents on the CIT fluorescence intensity. The results show that lower pH value, aceitc acid, β-CD and acetonitrile can induce a higher fluorescence intensity of CIT, but methanol or H 2 O has a decreasing effect on the fluorescence intensity of CIT. Findings in this study provide a theoretical basis for development of a high sensitivity fluorescence-based trace analysis for CIT detection. - Highlights: ► The effects of pH, solvents and β-CD on the fluorescence of citrinin are analyzed. ► [H] + , acetic acid, β-CD and acetonitrile can induce CIT fluorescence enhancement. ► Methanol and H 2 O can induce CIT fluorescence reduction. ► The 1:1 inclusion complex of CIT/β-CD can form in acidic phosphate solution.

Development of a reconstruction software of elemental maps by micro X-ray fluorescence

International Nuclear Information System (INIS)

Almeida, Andre Pereira de; Braz, Delson; Mota, Carla Lemos; Oliveira, Luis Fernando de; Barroso, Regina Cely; Pinto, Nivia Graciele Villela; Cardoso, Simone Coutinho; Moreira, Silvana

2009-01-01

The technique of X-ray fluorescence (XRF) using SR microbeams is a powerful analysis tool for studying elemental composition in several samples. One application of this technique is the analysis done through the mapping of chemical elements forming a matrix of data. The aim of this work is the presentation of the program MapXRF, an in-house software designed to optimize the processing and mapping of fluorescence intensities data. This program uses spectra generated by QXAS as input data and separates the intensities of each chemical element found in the fluorescence spectra in files themselves. From these files, the program generates the intensity maps that can be visualized in any program of treatment of images. The proposed software was tested using fluorescence data obtained in the XRF beamline at National Synchrotron Light Laboratory (LNLS), Brazil. Automatic 2D scans were performed and element distribution maps were obtained in form of a matrix of data. (author)

Development of a reconstruction software of elemental maps by micro X-ray fluorescence

Energy Technology Data Exchange (ETDEWEB)

Almeida, Andre Pereira de; Braz, Delson; Mota, Carla Lemos, E-mail: apalmeid@gmail.co, E-mail: delson@lin.ufrj.b, E-mail: clemos@con.ufrj.b [Universidade Federal do Rio de Janeiro (PEN/COPPE/UFRJ), RJ (Brazil). Coordenacao dos Programas de Pos-Graduacao de Engenharia. Programa de Energia Nuclear; Oliveira, Luis Fernando de; Barroso, Regina Cely; Pinto, Nivia Graciele Villela, E-mail: cely@uerj.b, E-mail: lfolive@uerj.b, E-mail: nitatag@gmail.co [Universidade do Estado do Rio de Janeiro (IF/UERJ), RJ (Brazil). Inst. de Fisica; Cardoso, Simone Coutinho [Universidade Federal do Rio de Janeiro (IF/UFRJ), RJ (Brazil). Inst. de Fisica; Moreira, Silvana [Universidade Estadual de Campinas (FEC/UNICAMP), SP (Brazil) Faculdade de Engenharia Civil, Arquitetura e Urbanismo

2009-07-01

The technique of X-ray fluorescence (XRF) using SR microbeams is a powerful analysis tool for studying elemental composition in several samples. One application of this technique is the analysis done through the mapping of chemical elements forming a matrix of data. The aim of this work is the presentation of the program MapXRF, an in-house software designed to optimize the processing and mapping of fluorescence intensities data. This program uses spectra generated by QXAS as input data and separates the intensities of each chemical element found in the fluorescence spectra in files themselves. From these files, the program generates the intensity maps that can be visualized in any program of treatment of images. The proposed software was tested using fluorescence data obtained in the XRF beamline at National Synchrotron Light Laboratory (LNLS), Brazil. Automatic 2D scans were performed and element distribution maps were obtained in form of a matrix of data. (author)

The fluorescence properties of aerosol larger than 0.8 μm in an urban and a PBA-dominated location

Science.gov (United States)

Gabey, A. M.; Stanley, W. R.; Gallagher, M. W.; Kaye, P. H.

2011-01-01

Dual-wavelength Ultraviolet light-induced fluorescence (UV-LIF) measurements were performed on ambient environmental aerosol in Manchester, UK (urban city centre, winter) and Borneo, Malaysia (remote, tropical), which are taken to represent environments with negligible and significant primary biological aerosol (PBA) influences, respectively. Single-particle fluorescence intensity and optical equivalent diameter were measured with a Wide Issue Bioaerosol Sensor, version 3 (WIBS3) in the diameter range 0.8 μm≤DP≤20 μm for 2-3 weeks and filters were analysed using energy dispersive X-ray (EDX) spectroscopy, which revealed mostly non-PBA dominated particle sizes larger than 1 μm in Manchester. The WIBS3 features three fluorescence channels: Fluorescence excited at 280 nm is recorded at 310-400 nm and 400-600 nm and fluorescence excited at 370 nm is detected at 400-600 nm. In Manchester the primary size mode of fluorescent and non-fluorescent material was at 1.2 μm. In Borneo non-fluorescent material peaked at 1.2 μm and fluorescent at 3-4 μm. The fluorescence intensity at 400-600 nm generally increased with DP at both sites, as did the 310-400 nm intensity in Borneo. In Manchester the 310-400 m fluorescence decreased at DP>4 μm, suggesting this channel offers additional discrimination between fluorescent particle types. Finally, the ratio of fluorescence intensity in two pairs of channels was investigated as a function of particle diameter and this varied significantly between the two environments, demonstrating that the fluorescent aerosol in each can in principle be distinguished using a combination of fluorescence and elastic scattering measurements.

Fluorescence image excited by a scanning UV-LED light

Science.gov (United States)

Tsai, Hsin-Yi; Chen, Yi-Ju; Huang, Kuo-Cheng

2013-03-01

An optical scanning system using UV-LED light to induced fluorescence technology can enhance a fluorescence image significantly in a short period. It has several advantages such as lower power consumption, no scattering effect in skins, and multilayer images can be obtained to analyze skin disease. From the experiment results, the light intensity increases with increase spot size and decrease scanning speed, but the image resolution is oppositely. Moreover, the system could be widely used in clinical diagnosis and photodynamic therapy for skin disease because even the irradiated time of fluorescence substance is short but it will provide accurately positioning of fluorescence object.

Selective recognition of Pr3+ based on fluorescence enhancement sensor

International Nuclear Information System (INIS)

Ganjali, M.R.; Hosseini, M.; Ghafarloo, A.; Khoobi, M.; Faridbod, F.; Shafiee, A.; Norouzi, P.

2013-01-01

(E)-2-(1-(4-hydroxy-2-oxo-2H-chromen-3-yl)ethylidene) hydrazinecarbothioamide (L) has been used to detect trace amounts of praseodymium ion in acetonitrile–water solution (MeCN/H 2 O) by fluorescence spectroscopy. The fluorescent probe undergoes fluorescent emission intensity enhancement upon binding to Pr 3+ ions in MeCN/H 2 O (9/1:v/v) solution. The fluorescence enhancement of L is attributed to a 1:1 complex formation between L and Pr 3+ , which has been utilized as the basis for selective detection of Pr 3+ . The sensor can be applied to the quantification of praseodymium ion with a linear range of 1.6 × 10 −7 to 1.0 × 10 −5 M. The limit of detection was 8.3 × 10 −8 M. The sensor exhibits high selectivity toward praseodymium ions in comparison with common metal ions. The proposed fluorescent sensor was successfully used for determination of Pr 3+ in water samples. - Highlights: • A new fluorescent sensor is introduced as a selective probe for Pr 3+ detection. • Fluorescent intensity of the chemical probe enhances upon binding to Pr 3+ ion. • The sensor can be used for Pr 3+ determination in the range of 1.6 × 10 −7 –1.0 × 10 −5 M

Determination of fission products in irradiated fuel by X-ray fluorescence

International Nuclear Information System (INIS)

Mogensen, M.; Als-Nielsen, J.; Hessel Andersen, N.

1986-08-01

X-ray fluorescence is a well established analytical tool for measuring elemental composition of fairly large (approximately 5 cm 2 ) ''cold'' samples. A version of this technique has been developed for analysis of radial distribution of fission products Xe, Cs and Ba in irradiated UO 2 fuel samples. About 0.1 mm thin slices of fuel pellets (full cross sections) are irradiated by 50 keV X-rays. The intensity of the Xe (Cs, Ba) K α fluorescence radiation generated is measured by means of a Ge detector fitted with a collimator. The slit is 0.5 mm wide in the scanning direction and 2 mm long. The measured Xe K α X-ray intensities are converted to absolute concentrations by comparing to the intensity from a Xe gas standard. In the case of Cs and Ba solid standards may be used. The X-ray fluorescence analysis is compared to other techniques used to obtain radial fission product profiles. It is shown how a combination of X-ray fluorescence and electron probe micro analysis is able to reveal the amount of Xe in the grain boundary porosities. (author)

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

Science.gov (United States)

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.

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

Science.gov (United States)

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

2015-12-01

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

Nonmydriatic fluorescence-based quantitative imaging of human macular pigment distributions

Science.gov (United States)

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

2006-10-01

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

An approach to estimate spatial distribution of analyte within cells using spectrally-resolved fluorescence microscopy

Science.gov (United States)

Sharma, Dharmendar Kumar; Irfanullah, Mir; Basu, Santanu Kumar; Madhu, Sheri; De, Suman; Jadhav, Sameer; Ravikanth, Mangalampalli; Chowdhury, Arindam

2017-03-01

While fluorescence microscopy has become an essential tool amongst chemists and biologists for the detection of various analyte within cellular environments, non-uniform spatial distribution of sensors within cells often restricts extraction of reliable information on relative abundance of analytes in different subcellular regions. As an alternative to existing sensing methodologies such as ratiometric or FRET imaging, where relative proportion of analyte with respect to the sensor can be obtained within cells, we propose a methodology using spectrally-resolved fluorescence microscopy, via which both the relative abundance of sensor as well as their relative proportion with respect to the analyte can be simultaneously extracted for local subcellular regions. This method is exemplified using a BODIPY sensor, capable of detecting mercury ions within cellular environments, characterized by spectral blue-shift and concurrent enhancement of emission intensity. Spectral emission envelopes collected from sub-microscopic regions allowed us to compare the shift in transition energies as well as integrated emission intensities within various intracellular regions. Construction of a 2D scatter plot using spectral shifts and emission intensities, which depend on the relative amount of analyte with respect to sensor and the approximate local amounts of the probe, respectively, enabled qualitative extraction of relative abundance of analyte in various local regions within a single cell as well as amongst different cells. Although the comparisons remain semi-quantitative, this approach involving analysis of multiple spectral parameters opens up an alternative way to extract spatial distribution of analyte in heterogeneous systems. The proposed method would be especially relevant for fluorescent probes that undergo relatively nominal shift in transition energies compared to their emission bandwidths, which often restricts their usage for quantitative ratiometric imaging in

Fluorescence recognition of chiral amino alcohols by using a novel ionic liquid sensor.

Science.gov (United States)

Cai, Pengfei; Wu, Datong; Zhao, Xiaoyong; Pan, Yuanjiang

2017-08-07

A novel task-specific ionic liquid derived from l-phenylalaninol was prepared as an enantioselective fluorescent sensor for the first time. Fluorescent chiral ionic liquid 1 (FCIL1) is found to exhibit highly enantioselective fluorescence enhancements toward both aromatic and non-aromatic chiral amino alcohols. When (S)-FCIL1 was treated with the enantiomers of phenylalaninol, a great fluorescence enhancement at 349 nm could be observed and the value of the enantiomeric fluorescence difference (ef) is 5.92. This demonstrated that the chiral sensor (S)-FCIL1 exhibited an excellent enantioselective response behaviour to d-phenylalaninol. Besides that, both the fluorescence intensity at 349 nm (I 349 ) and the ratio of I 349 to I 282 depend linearly on the concentration of amino alcohols. Both the concentration and the enantiomeric composition could be determined by using the chiral ionic liquid. Differently, the sensor treated with the enantiomers of 2-amino-1-butanol showed an opposite result: the fluorescence intensity of the S-enantiomer is higher than that of the R-enantiomer. Furthermore, the size of the substituents on the chiral carbon might be important for the enantioselective fluorescent response.

Highly selective ratiometric fluorescent detection of Fe{sup 3+} with a polyphenyl derivative

Energy Technology Data Exchange (ETDEWEB)

Li, Zhan-Xian, E-mail: lizx@zzu.edu.cn [The College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Zhou, Wan; Zhang, Li-Feng; Yuan, Rui-Li; Liu, Xing-Jiang; Wei, Liu-He [The College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China); Yu, Ming-Ming, E-mail: yumm@zzu.edu.cn [The College of Chemistry and Molecular Engineering, Zhengzhou University, Zhengzhou 450001 (China)

2013-04-15

Compared with other fluorescent probes, ratiometric fluorescence responses are more attractive because the ratio between the two emission intensities can be used to measure the analyte concentration and provide a built-in correction for environmental effects. A highly selective and sensitive ratiometric fluorescent probe for Fe{sup 3+} was synthesized, which exhibits an enhanced fluorescence with a large red-shift in emission from 361 to 455 nm upon addition of Fe{sup 3+}. The red-shift of the emission peak can be ascribed to the reformed orbital, and the increase of emission intensity may be ascribed to the inhibition of the rotation of C–C bonds between each two aromatic rings. -- Graphical abstract: A highly selective and sensitive ratiometric fluorescent probe for Fe{sup 3+} was synthesized, which exhibits an enhanced fluorescence with a large red-shift in emission from 361 to 455 nm upon addition of Fe{sup 3+}. Highlights: ► A ratiometric fluorescent probe for Fe{sup 3+} was synthesized. ► The probe exhibits an enhanced fluorescence with a red-shift upon addition of Fe{sup 3+}. ► Inhibition of the rotation of C–C bonds was possible detection mechanism for Fe{sup 3+}.

Fluorescence Resonance Energy Transfer in Polydiacetylene Liposomes

Science.gov (United States)

Li, Xuelian; Matthews, Shelton; Kohli, Punit

2009-01-01

Conjugated polydiacetylene (PDA) possessing stimuli-responsive properties has been intensively investigated for developing efficient sensors. We report here fluorescence resonance energy transfer (FRET) in liposomes synthesized using different molar ratios of dansyl-tagged diacetylene and diacetylene–carboxylic acid monomers. Photopolymerization of diacetylene resulted in cross-linked PDA liposomes. We used steady-state electronic absorption, emission, and fluorescence anisotropy (FA) analysis to characterize the thermal-induced FRET between dansyl fluorophores (donor) and PDA (acceptor). We found that the monomer ratio of acceptor to donor (Rad) and length of linkers (functional part that connects dansyl fluorophores to the diacetylene group in the monomer) strongly affected FRET. For Rad = 10 000, the acceptor emission intensity was amplified by more than 18 times when the liposome solution was heated from 298 to 338 K. A decrease in Rad resulted in diminished acceptor emission amplification. This was primarily attributed to lower FRET efficiency between donors and acceptors and a higher background signal. We also found that the FRET amplification of PDA emissions after heating the solution was much higher when dansyl was linked to diacetylene through longer and flexible linkers than through shorter linkers. We attributed this to insertion of dansyl in the bilayer of the liposomes, which led to an increased dansyl quantum yield and a higher interaction of multiple acceptors with limited available donors. This was not the case for shorter and more rigid linkers where PDA amplification was much smaller. The present studies aim at enhancing our understanding of FRET between fluorophores and PDA-based conjugated liposomes. Furthermore, receptor tagged onto PDA liposomes can interact with ligands present on proteins, enzymes, and cells, which will produce emission sensing signal. Therefore, using the present approach, there exist opportunities for designing FRET

Platinum plasmonic nanostructure arrays for massively parallel single-molecule detection based on enhanced fluorescence measurements

International Nuclear Information System (INIS)

Saito, Toshiro; Takahashi, Satoshi; Obara, Takayuki; Itabashi, Naoshi; Imai, Kazumichi

2011-01-01

We fabricated platinum bowtie nanostructure arrays producing fluorescence enhancement and evaluated their performance using two-photon photoluminescence and single-molecule fluorescence measurements. A comprehensive selection of suitable materials was explored by electromagnetic simulation and Pt was chosen as the plasmonic material for visible light excitation near 500 nm, which is preferable for multicolor dye-labeling applications like DNA sequencing. The observation of bright photoluminescence (λ = 500-600 nm) from each Pt nanostructure, induced by irradiation at 800 nm with a femtosecond laser pulse, clearly indicates that a highly enhanced local field is created near the Pt nanostructure. The attachment of a single dye molecule was attempted between the Pt triangles of each nanostructure by using selective immobilization chemistry. The fluorescence intensities of the single dye molecule localized on the nanostructures were measured. A highly enhanced fluorescence, which was increased by a factor of 30, was observed. The two-photon photoluminescence intensity and fluorescence intensity showed qualitatively consistent gap size dependence. However, the average fluorescence enhancement factor was rather repressed even in the nanostructure with the smallest gap size compared to the large growth of photoluminescence. The variation of the position of the dye molecule attached to the nanostructure may influence the wide distribution of the fluorescence enhancement factor and cause the rather small average value of the fluorescence enhancement factor.

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

Science.gov (United States)

Hou, Faju; Wang, Xiaolei; Jiang, Chongqiu

2005-03-01

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

X-ray luminescence computed tomography imaging via multiple intensity weighted narrow beam irradiation

Science.gov (United States)

Feng, Bo; Gao, Feng; Zhao, Huijuan; Zhang, Limin; Li, Jiao; Zhou, Zhongxing

2018-02-01

The purpose of this work is to introduce and study a novel x-ray beam irradiation pattern for X-ray Luminescence Computed Tomography (XLCT), termed multiple intensity-weighted narrow-beam irradiation. The proposed XLCT imaging method is studied through simulations of x-ray and diffuse lights propagation. The emitted optical photons from X-ray excitable nanophosphors were collected by optical fiber bundles from the right-side surface of the phantom. The implementation of image reconstruction is based on the simulated measurements from 6 or 12 angular projections in terms of 3 or 5 x-ray beams scanning mode. The proposed XLCT imaging method is compared against the constant intensity weighted narrow-beam XLCT. From the reconstructed XLCT images, we found that the Dice similarity and quantitative ratio of targets have a certain degree of improvement. The results demonstrated that the proposed method can offer simultaneously high image quality and fast image acquisition.

Fluorescence Enhancement of Fluorescent Unnatural Streptavidin by Binding of a Biotin Analogue with Spacer Tail and Its Application to Biotin Sensing

Directory of Open Access Journals (Sweden)

Xianwei Zhu

2014-01-01

Full Text Available We designed a novel molecular biosensing system for the detection of biotin, an important vitamin by the combination of fluorescent unnatural streptavidin with a commercialized biotin-(AC52-hydrazide. A fluorescent unnatural amino acid, BODIPY-FL-aminophenylalanine (BFLAF, was position-specifically incorporated into Trp120 of streptavidin by four-base codon method. Fluorescence of the Trp120BFLAF mutant streptavidin was enhanced by the addition of biotin-(AC52-hydrazide with the concentration dependent, whereas fluorescence enhancement was not observed at all by the addition of natural biotin. It was considered that the spacer tail of biotin-(AC52-hydrazide may disturb the fluorescence quenching of the Trp120BFLAF by Trp79 and Trp108 of the neighbor subunit. Therefore, biotin sensing was carried out by the competitive binding reaction of biotin-(AC52-hydrazide and natural biotin to the fluorescent mutant streptavidin. The fluorescence intensity decreased by increasing free biotin concentration. The result suggested that molecular biosensor for small ligand could be successfully designed by the pair of fluorescent mutant binding protein and ligand analogue.

Fluorescent IgG fusion proteins made in E. coli

Science.gov (United States)

Luria, Yael; Raichlin, Dina; Benhar, Itai

2012-01-01

Antibodies are among the most powerful tools in biological and biomedical research and are presently the fastest growing category of new bio-pharmaceutics. The most common format of antibody applied for therapeutic, diagnostic and analytical purposes is the IgG format. For medical applications, recombinant IgGs are made in cultured mammalian cells in a process that is too expensive to be considered for producing antibodies for diagnostic and analytical purposes. Therefore, for such purposes, mouse monoclonal antibodies or polyclonal sera from immunized animals are used. While looking for an easier and more rapid way to prepare full-length IgGs for therapeutic purposes, we recently developed and reported an expression and purification protocol for full-length IgGs, and IgG-based fusion proteins in E. coli, called “Inclonals.” By applying the Inclonals technology, we could generate full-length IgGs that are genetically fused to toxins. The aim of the study described herein was to evaluate the possibility of applying the “Inclonals” technology for preparing IgG-fluorophore fusion proteins. We found that IgG fused to the green fluorescent proteins enhanced GFP (EGFP) while maintaining functionality in binding, lost most of its fluorescence during the refolding process. In contrast, we found that green fluorescent Superfolder GFP (SFGFP)-fused IgG and red fluorescent mCherry-fused IgG were functional in antigen binding and maintained fluorescence intensity. In addition, we found that we can link several SFGFPs in tandem to each IgG, with fluorescence intensity increasing accordingly. Fluorescent IgGs made in E. coli may become attractive alternatives to monoclonal or polyclonal fluorescent antibodies derived from animals. PMID:22531449

The Synthesis of Wavelength-Controlled CdTe/Hydroxyapatite Composites and Their Fluorescence Enhancement by Bovine Serum Albumin

Directory of Open Access Journals (Sweden)

Li Jin

2016-01-01

Full Text Available For the last ten years, quantum dots modified by biological materials have made it possible to study biochemical processes by means of biomedical imaging. This thesis introduced how the fluorescence CdTe quantum dots/hydroxyapatite composites were synthesized and how their structure, morphology, and fluorescence property were characterized by using TEM, XRD, EDS, UV-vis absorption spectra, and fluorescence spectra. The fluorescence spectra indicated the superb photometric characteristics of CdTe/HA composites. We also found that refluxing temperature and time had prominent effects on fluorescence wavelength and intensity of CdTe/HA composites, so the fluorescence emission wavelength of CdTe/HA composites could be controlled. In addition, the effect of BSA on the fluorescence properties of CdTe/HA composites was studied. The fluorescent emission intensity of CdTe/HA composites was enhanced directly with increasing concentrations of BSA; meanwhile, the fluorescence emission intensity of BSA dramatically decreased, which indicated that a Förster nonradiative energy transfer process occurred through the formation of chemical bonds between BSA and CdTe/HA composites. And the two-dimensional correlation (2D COS was used to analyze the BSA solution before and after the reaction, which indicated that CdTe/HA composites have bound to a site at the surface of the molecule in the first subdomain IA. We also found that there was a linear relationship between the fluorescence intensity enhancement (F/F0 of CdTe/HA composites and the concentration of the bovine serum albumin, which might become a method for quantitative analysis of BSA in a real sample.

Multispectral fluorescence image algorithms for detection of frass on mature tomatoes

Science.gov (United States)

A multispectral algorithm derived from hyperspectral line-scan fluorescence imaging under violet LED excitation was developed for the detection of frass contamination on mature tomatoes. The algorithm utilized the fluorescence intensities at five wavebands, 515 nm, 640 nm, 664 nm, 690 nm, and 724 nm...

Image recovery from defocused 2D fluorescent images in multimodal digital holographic microscopy.

Science.gov (United States)

Quan, Xiangyu; Matoba, Osamu; Awatsuji, Yasuhiro

2017-05-01

A technique of three-dimensional (3D) intensity retrieval from defocused, two-dimensional (2D) fluorescent images in the multimodal digital holographic microscopy (DHM) is proposed. In the multimodal DHM, 3D phase and 2D fluorescence distributions are obtained simultaneously by an integrated system of an off-axis DHM and a conventional epifluorescence microscopy, respectively. This gives us more information of the target; however, defocused fluorescent images are observed due to the short depth of field. In this Letter, we propose a method to recover the defocused images based on the phase compensation and backpropagation from the defocused plane to the focused plane using the distance information that is obtained from a 3D phase distribution. By applying Zernike polynomial phase correction, we brought back the fluorescence intensity to the focused imaging planes. The experimental demonstration using fluorescent beads is presented, and the expected applications are suggested.

Role of fluorescence study in diagnosis of reccurent skin cancer (case report

Directory of Open Access Journals (Sweden)

E. V. Filonenko

2013-01-01

Full Text Available The case of diagnosis of recurrent skin cancer using fluorescence methods is reported. The patient who underwent multiple courses of treatment (surgical resection, cryotherapy and laser ablation for multifocal basal cell carcinoma of head. trunk and extremities. The patient represented with multiple whitish scars in the region of surgery, cryotherapy and laser ablation (one of them was in the region of external ear and with multiple foci of hyperemia and skin peeling. The patient underwent fluorescence diagnosis according to developed regimen: alasens was given per os at a dose of 30 mg/kg body weight 4 h before study. In the white light there were no features of recurrent tumor in scar areas. During fluorescence examination there was bright red fluorescence in the area of ear scar, which was intact visually, with no additional fluorescence foci in regions of surgical resection, cryotherapy and laser ablation and other regions. According to cytological study of shave biopsy from fluorescecnse area of ear scar the recurrence of basal cell skin carcinoma was diagnosed. 

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

Science.gov (United States)

Lee, Ji-Won; Iimura, Tadahiro

2017-02-01

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

Fluorescent determination of neptunium in plutonium

International Nuclear Information System (INIS)

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

1991-01-01

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

Fluorescent excitation of interstellar H2

NARCIS (Netherlands)

Black, J.H.; Dishoeck, van E.F.

1987-01-01

The infrared emission spectrum of H2 excited by ultraviolet absorption, followed by fluorescence, was investigated using comprehensive models of interstellar clouds for computing the spectrum and to assess the effects on the intensity to various cloud properties, such as density, size, temperature,

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

Energy Technology Data Exchange (ETDEWEB)

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

1976-07-01

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

A novel turn-on fluorescent strategy for sensing ascorbic acid using graphene quantum dots as fluorescent probe.

Science.gov (United States)

Liu, Hua; Na, Weidan; Liu, Ziping; Chen, Xueqian; Su, Xingguang

2017-06-15

In this paper, a facile and rapid fluorescence turn-on assay for fluorescent detection of ascorbic acid (AA) was developed by using the orange emission graphene quantum dots (GQDs). In the presence of horse radish peroxidase (HRP) and hydrogen peroxide (H 2 O 2 ), catechol can be oxidized by hydroxyl radicals and converted to o-benzoquinone, which can significantly quench the fluorescence of GQDs. However, when AA present in the system, it can consume part of H 2 O 2 and hydroxyl radicals to inhibit the generation of o-benzoquinone, resulting in fluorescence recovery. Under the optimized experimental conditions, the fluorescence intensity was linearly correlated with the concentration of H 2 O 2 in the range of 3.33-500µM with a detection limit of 1.2µM. The linear detection for AA was in the range from 1.11 to 300µM with a detection limit of 0.32µM. The proposed method was applied to the determination of AA in human serum samples with satisfactory results. Copyright © 2017. Published by Elsevier B.V.

Fluorescence optical imaging in anticancer drug delivery.

Science.gov (United States)

Etrych, Tomáš; Lucas, Henrike; Janoušková, Olga; Chytil, Petr; Mueller, Thomas; Mäder, Karsten

2016-03-28

In the past several decades, nanosized drug delivery systems with various targeting functions and controlled drug release capabilities inside targeted tissues or cells have been intensively studied. Understanding their pharmacokinetic properties is crucial for the successful transition of this research into clinical practice. Among others, fluorescence imaging has become one of the most commonly used imaging tools in pre-clinical research. The development of increasing numbers of suitable fluorescent dyes excitable in the visible to near-infrared wavelengths of the spectrum has significantly expanded the applicability of fluorescence imaging. This paper focuses on the potential applications and limitations of non-invasive imaging techniques in the field of drug delivery, especially in anticancer therapy. Fluorescent imaging at both the cellular and systemic levels is discussed in detail. Additionally, we explore the possibility for simultaneous treatment and imaging using theranostics and combinations of different imaging techniques, e.g., fluorescence imaging with computed tomography. Copyright © 2016 Elsevier B.V. All rights reserved.

Development of a fluorescent chelating ligand for scandium ion having a Schiff base moiety

Science.gov (United States)

Yamada, Hiroshi; Kojo, Masahito; Nakahara, Tomomi; Murakami, Kumi; Kakima, Takashi; Ichiba, Hideaki; Yajima, Takehiko; Fukushima, Takeshi

2012-05-01

A fluorescent ligand, 1-(2-hydroxy-3-methoxybenzaldehyde)-4-aminosalicylhydrazone (HMB-ASH), was newly designed and synthesized, and its fluorescence characteristics for metal ions were investigated in the pH range 3.0-10.5 (at a difference of 0.5 for each metal). After testing 31 different metal ions, it was found that HMB-ASH was able to emit fluorescence intensely at 512 nm with an excitation wavelength of 353 nm in the presence of Sc3+, one of the rare earth metals, at pH values around 3.5 and 8.0. The other metal ions hardly showed fluorescence with HMB-ASH. The fluorescence was more intense at pH 8.0, and the detection limit of Sc3+ in a buffer solution (pH 8.0) was approximately 18.8 nmol L-1 (0.85 ppb).

Study on excitation and fluorescence spectrums of Japanese citruses to construct machine vision systems for acquiring fluorescent images

Science.gov (United States)

Momin, Md. Abdul; Kondo, Naoshi; Kuramoto, Makoto; Ogawa, Yuichi; Shigi, Tomoo

2011-06-01

Research was conducted to acquire knowledge of the ultraviolet and visible spectrums from 300 -800 nm of some common varieties of Japanese citrus, to investigate the best wave-lengths for fluorescence excitation and the resulting fluorescence wave-lengths and to provide a scientific background for the best quality fluorescent imaging technique for detecting surface defects of citrus. A Hitachi U-4000 PC-based microprocessor controlled spectrophotometer was used to measure the absorption spectrum and a Hitachi F-4500 spectrophotometer was used for the fluorescence and excitation spectrums. We analyzed the spectrums and the selected varieties of citrus were categorized into four groups of known fluorescence level, namely strong, medium, weak and no fluorescence.The level of fluorescence of each variety was also examined by using machine vision system. We found that around 340-380 nm LEDs or UV lamps are appropriate as lighting devices for acquiring the best quality fluorescent image of the citrus varieties to examine their fluorescence intensity. Therefore an image acquisition device was constructed with three different lighting panels with UV LED at peak 365 nm, Blacklight blue lamps (BLB) peak at 350 nm and UV-B lamps at peak 306 nm. The results from fluorescent images also revealed that the findings of the measured spectrums worked properly and can be used for practical applications such as for detecting rotten, injured or damaged parts of a wide variety of citrus.

The fluorescence properties of aerosol larger than 0.8 μm in urban and tropical rainforest locations

Science.gov (United States)

Gabey, A. M.; Stanley, W. R.; Gallagher, M. W.; Kaye, P. H.

2011-06-01

UV-LIF measurements were performed on ambient aerosol in Manchester, UK (urban city centre, winter) and Borneo, Malaysia (remote, tropical) using a Wide Issue Bioaerosol Spectrometer, version 3 (WIBS3). These sites are taken to represent environments with minor and significant primary biological aerosol (PBA) influences respectively, and the urban dataset describes the fluorescent background aerosol against which PBA must be identified by researchers using LIF. The ensemble aerosol at both sites was characterised over 2-3 weeks by measuring the fluorescence intensity and optical equivalent diameter (DP) of single particles sized 0.8 ≤ DP ≤ 20 μm. Filter samples were also collected for a subset of the Manchester campaign and analysed using energy dispersive X-Ray (EDX) spectroscopy and environmental scanning electron microscopy (ESEM), which revealed mostly non-PBA at D ≤ 1 μm. The WIBS3 features three fluorescence channels: the emission following a 280 nm excitation is recorded at 310-400 nm (channel F1) and 400-600 nm (F2), and fluorescence excited at 350 nm is detected at 400-600 nm (F3). In Manchester the primary size mode of fluorescent and non-fluorescent material was present at 0.8-1.2 μm, with a secondary fluorescent mode at 2-4 μm. In Borneo non-fluorescent material peaked at 0.8-1.2 μm and fluorescent at 3-4 μm. Agreement between fluorescent number concentrations in each channel differed at the two sites, with F1 and F3 reporting similar concentrations in Borneo but F3 outnumbering F1 by a factor of 2-3 across the size spectrum in Manchester. The fluorescence intensity in each channel generally rose with DP at both sites with the exception of F1 intensity in Manchester, which peaked at DP = 4 μm, causing a divergence between F1 and F3 intensity at larger DP. This divergence and the differing fluorescent particle concentrations demonstrate the additional discrimination provided by the F1 channel in Manchester. The relationships between

Detection of Sn(II) ions via quenching of the fluorescence of carbon nanodots

International Nuclear Information System (INIS)

Mohd Yazid, S.N.A.; Chin, S.F.; Pang, S.C.; Ng, S.M.

2013-01-01

We report that fluorescent carbon nanodots (C-dots) can act as an optical probe for quantifying Sn(II) ions in aqueous solution. C-dots are synthesized by carbonization and surface oxidation of preformed sago starch nanoparticles. Their fluorescence is significantly quenched by Sn(II) ions, and the effect can be used to determine Sn(II) ions. The highest fluorescence intensity is obtained at a concentration of 1.75 mM of C-dots in aqueous solution. The probe is highly selective and hardly interfered by other ions. The quenching mechanism appears to be predominantly of the static (rather than dynamic) type. Under optimum conditions, there is a linear relationship between fluorescence intensity and Sn(II) ions concentration up to 4 mM, and with a detection limit of 0.36 μM. (author)

Development of a fluorescent x-ray source for medical imaging

Science.gov (United States)

Toyofuku, F.; Tokumori, K.; Nishimura, K.; Saito, T.; Takeda, T.; Itai, Y.; Hyodo, K.; Ando, M.; Endo, M.; Naito, H.; Uyama, C.

1995-02-01

A fluorescent x-ray source for medical imaging, such as K-edge subtraction angiography and monochromatic x-ray CT, has been developed. Using a 6.5 GeV accumulation ring in Tsukuba, fluorescent x rays, which range from about 30 to 70 keV are generated by irradiating several target materials. Measurements have been made of output intensities and energy spectra for different target angles and extraction angles. The intensities of fluorescent x rays at a 30 mA beam current are on the order of 1-3×106 photons/mm2/s at 30 cm from the local spot where the incident beam is collimated to 1 mm2. A phantom which contains three different contrast media (iodine, barium, gadolinium) was used for the K-edge energy subtraction, and element selective CT images were obtained.

[Simultaneous determination of multiple elements in airborne particulate samples by X-ray fluorescence spectrometry].

Science.gov (United States)

Takada, T; Hitosugi, M; Kadowaki, T; Kudo, M

1983-07-01

An energy dispersive X-ray fluorescence spectrometer (EDX) has been applied to determine multielements in the workplace air. The standards for X-ray fluorescence analysis were prepared by the chelate precipitation method on polyvinyl chloride (PVC) membrane filter. And, the specimens were prepared to deposit various metal compounds of different chemical forms by the suspension method on PVC membrane filter, and they were determined with EDX and atomic absorption spectrometer (AAS). The results obtained were as follows. Though there is a difference by each element, an amount less than 3 microgram/cm2 per unit area makes it possible to undergo multielement analysis, that is, is has no influence on fine particle effect (particle size; under 5 microns). Then, effects of the X-ray intensity by different chemical forms are negligible. At the presence the neighboring element and other elements this technique showed greater precision by carrying out on corrective treatment, etc. The coefficient of variation of this technique was in the range of 2.5-6.5% at DDTC-Cu of 0.5-5.0 micrograms/cm2, with the limit of detection for As : 0.002 microgram/cm2, Zn : 0.003 microgram/cm2, Pb : 0.003 microgram/cm2, Cu : 0.004 microgram/cm2, Ni : 0.003 microgram/cm2, Fe : 0.005 microgram/cm2, Mn : 0.008 microgram/cm2, Cr : 0.013 microgram/cm2, respectively. Aerosols collected at the workplace were analyzed with EDX and AAS, and the obtained results showed good agreement with such regression line as y = 1.04 chi + 0.04, the coefficient of correlation being r = 0.995. From these results, this technique was found to be a very excellent method for monitoring of multielements in the workplace air.

Fluorescence-based calculus detection using a 405-nm excitation wavelength

Science.gov (United States)

Brede, O.; Schelle, F.; Krueger, S.; Oehme, B.; Dehn, C.; Frentzen, M.; Braun, A.

2011-03-01

The aim of this study was to assess the difference of fluorescence signals of cement and calculus using a 405 nm excitation wavelength. A total number of 20 freshly extracted teeth was used. The light source used for this study was a blue LED with a wavelength of 405nm. For each tooth the spectra of calculus and cementum were measured separately. Fluorescence light was collimated into an optical fibre and spectrally analyzed using an echelle spectrometer (aryelle 200, Lasertechnik Berlin, Germany) with an additionally bandpass (fgb 67, Edmund Industrial Optics, Karlsruhe, Germany). From these 40 measurements the median values were calculated over the whole spectrum, leading to two different median spectra, one for calculus and one for cementum. For further statistical analysis we defined 8 areas of interest (AOI) in wavelength regions, showing remarkable differences in signal strength. In 7 AOIs the intensity of the calculus spectrum differed statistically significant from the intensity of the cementum spectrum (p calculus and cement between 600nm and 700nm. Thus, we can conclude that fluorescence of calculus shows a significant difference to the fluorescence of cement. A differentiation over the intensity is possible as well as over the spectrum. Using a wavelength of 405nm, it is possible to distinguish between calculus and cement. These results could be used for further devices to develop a method for feedback controlled calculus removal.

Specific detection of Vibrio parahaemolyticus by fluorescence quenching immunoassay based on quantum dots.

Science.gov (United States)

Wang, Ling; Zhang, Junxian; Bai, Haili; Li, Xuan; Lv, Pintian; Guo, Ailing

2014-07-01

In this study, anti-Vibrio parahaemolyticus polyclonal and monoclonal antibodies were prepared through intradermal injection immune and lymphocyte hybridoma technique respectively. CdTe quantum dots (QDs) were synthesized at pH 9.3, 98 °C for 1 h with stabilizer of 2.7:1. The fluorescence intensity was 586.499, and the yield was 62.43%. QD probes were successfully prepared under the optimized conditions of pH 7.4, 37 °C for 1 h, 250 μL of 50 mg/mL EDC · HCl, 150 μL of 4 mg/mL NHS, buffer system of Na2HPO4-citric acid, and 8 μL of 2.48 mg/mL polyclonal antibodies. As gold nanoparticles could quench fluorescence of quantum dots, the concentration of V. parahaemolyticus could be detected through measuring the reduction of fluorescence intensity in immune sandwich reaction composed of quantum dot probe, gold-labeled antibody, and the sample. For pure culture, fluorescence intensity of the system was proportional with logarithm concentration of antigen, and the correlation coefficient was 99.764%. The fluorescence quenching immunoassay based on quantum dots is established for the first time to detect Vibrio parahaemolyticus. This method may be used as rapid testing procedure due to its high simplicity and sensitivity.

Copper nanoclusters as probes for turn-on fluorescence sensing of L-lysine.

Science.gov (United States)

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.

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

Science.gov (United States)

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

2013-06-01

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

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

International Nuclear Information System (INIS)

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

1994-01-01

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

Fluorescent metal nanoshell and CK19 detection on single cell image

International Nuclear Information System (INIS)

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

2011-01-01

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

A facile fluorescent sensor based on silicon nanowires for dithionite

Science.gov (United States)

Cao, Xingxing; Mu, Lixuan; Chen, Min; She, Guangwei

2018-05-01

A facile and novel fluorescent sensor for dithionite has been constructed by simultaneously immobilizing dansyl group (fluorescence molecule) and dabsyl group (quencher and recognizing group) on the silicon nanowires (SiNWs) and SiNW arrays surface. This sensor for dithionite exhibited high selectivity and a good relationship of linearity between fluorescence intensities and dithionite concentrations from 0.1 to 1 mM. This approach is straightforward and does not require complicated synthesis, which can be extended to develop other sensors with similar rationale.

Fluorescent multiple staining and CASA system to assess boar sperm viability and membranes integrity in short and long-term extenders

OpenAIRE

F. Cremonesi; A. Meucci; A. Lange-Consiglio

2013-01-01

The aim of this study was to assess the effect on boar spermatozoa quality of in vitro storage in short and long-term extenders by fluorescent multiple staining (FMS) and computer assisted semen analyzer (CASA). Fresh ejaculates from three healthy, sexually mature boars were diluted with equal volumes of six short-term or three long-term commercial extenders and stored at 19?C for 6 days (short-term) or 12 days (long-term). The integrity of spermatozoa membranes was analyzed by FMS using prop...

Using the geometric mean fluorescence intensity index method to measure ZAP-70 expression in patients with chronic lymphocytic leukemia

Directory of Open Access Journals (Sweden)

Wu YJ

2016-02-01

Full Text Available Yu-Jie Wu, Hui Wang, Jian-Hua Liang, Yi Miao, Lu Liu, Hai-Rong Qiu, Chun Qiao, Rong Wang, Jian-Yong Li Department of Hematology, First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, People’s Republic of China Abstract: Expression of ζ-chain-associated protein kinase 70 kDa (ZAP-70 in chronic lymphocytic leukemia (CLL is associated with more aggressive disease and can help differentiate CLL from cases expressing mutated or unmutated immunoglobulin heavy chain variable region (IgHV genes. However, standardizing ZAP-70 expression by flow cytometric analysis has proved unsatisfactory. The key point is that ZAP-70 is weakly expressed with a continuous expression pattern rather than a clear discrimination between positive and negative CLL cells, which means that the resulting judgment is subjective. Thus, in this study, we aimed at assessing the reliability and repeatability of ZAP-70 expression using the geometric mean fluorescence intensity (geo MFI index method based on flow cytometry with 256-channel resolution in a series of 402 CLL patients and to compare ZAP-70 with other biological and clinical prognosticators. According to IgHV mutational status, we were able to confirm that the optimal cut-off point for the geo MFI index was 3.5 in the test set. In multivariate analyses that included the major clinical and biological prognostic markers for CLL, the prognostic impact of ZAP-70 expression appeared to have stronger discriminatory power when the geo MFI index method was applied. In addition, we found that ZAP-70-positive patients according to the geo MFI index method had shorter time to first treatment or overall survival (P=0.0002, P=0.0491. This is the first report showing that ZAP-70 expression can be evaluated by a new approach, the geo MFI index, which could be a useful prognostic method as it is more reliable, less subjective, and therefore better associated with improvement of CLL prognostication

Synthesis and validation of novel cholesterol-based fluorescent lipids designed to observe the cellular trafficking of cationic liposomes.

Science.gov (United States)

Kim, Bieong-Kil; Seu, Young-Bae; Choi, Jong-Soo; Park, Jong-Won; Doh, Kyung-Oh

2015-09-15

Cholesterol-based fluorescent lipids with ether linker were synthesized using NBD (Chol-E-NBD) or Rhodamine B (Chol-E-Rh), and the usefulnesses as fluorescent probes for tracing cholesterol-based liposomes were validated. The fluorescent intensities of liposomes containing these modified lipids were measured and observed under a microscope. Neither compound interfered with the expression of GFP plasmid, and live cell images were obtained without interferences. Changes in the fluorescent intensity of liposomes containing Chol-E-NBD were followed by flow cytometry for up to 24h. These fluorescent lipids could be useful probes for trafficking of cationic liposome-mediated gene delivery. Copyright © 2015 Elsevier Ltd. All rights reserved.

Towards understanding of poly-guanine activated fluorescent silver nanoclusters

International Nuclear Information System (INIS)

Walczak, Sylwia; Morishita, Kiyoshi; Ahmed, Moin; Liu, Juewen

2014-01-01

It has been recently reported that the fluorescence of some DNA-templated silver nanoclusters (AgNCs) can be significantly enhanced upon by hybridizing with a partially complementary DNA containing a G-rich overhang near the AgNCs. This discovery has found a number of analytical applications but many fundamental questions remain to be answered. In this work, the photostability of these activated AgNCs is reported. After adding the G-rich DNA activator, the fluorescence intensity peaks in ∼1 h and then starts to decay, where the decaying rate is much faster with light exposure. The lost fluorescence is recovered by adding NaBH 4 , suggesting that the bleaching is an oxidative process. Once activated, the G-rich activator can be removed while the AgNCs still maintain most of their fluorescence intensity. UV–vis spectroscopy suggests that new AgNC species are generated upon hybridization with the activator. The base sequence and length of the template DNA have also been varied, leading to different emission colors and color change after hybridization. G-rich aptamers can also serve as activators. Our results indicate that activation of the fluorescence by G-rich DNA could be a convenient method for biosensor development since the unstable NaBH 4 is not required for the activation step. (paper)

Spectrally resolved pressure dependence measurements of air fluorescence emission with AIRFLY

International Nuclear Information System (INIS)

Ave, M.; Bohacova, M.; Buonomo, B.; Busca, N.; Cazon, L.; Chemerisov, S.D.; Conde, M.E.; Crowell, R.A.; Di Carlo, P.; Di Giulio, C.; Doubrava, M.; Esposito, A.; Facal, P.; Franchini, F.J.; Hoerandel, J.; Hrabovsky, M.; Iarlori, M.; Kasprzyk, T.E.; Keilhauer, B.; Klages, H.

2008-01-01

The knowledge of the fluorescence emission as a function of atmospheric parameters is essential for the detection of extensive air showers with the fluorescence technique. In this paper, we summarize AIRFLY published measurements of the pressure dependence of the fluorescence yield. The spectral distribution of the fluorescent light between 280 and 429 nm has been measured with high resolution. Relative intensities of 34 spectral lines have been determined. The pressure dependence of 25 lines was measured in terms of quenching reference pressures p λ ' in air. This set of AIRFLY measurements yields the most comprehensive parametrization of the pressure dependence of the fluorescent spectrum.

pH-responsive fluorescence chemical sensor constituted by conjugated polymers containing pyridine rings.

Science.gov (United States)

Adachi, Naoya; Kaneko, Yuki; Sekiguchi, Kazuki; Sugiyama, Hiroki; Sugeno, Masafumi

2015-12-01

Poly(p-pyridinium phenylene ethynylene)s (PPyPE) functionalized with alternating donor-acceptor repeat units were synthesized by a Pd-catalyzed Sonogashira coupling reaction between diethynyl monomer and di-iodopyridine for use as a pH-responsive fluorescence chemical sensor. The synthesized PPyPE, containing pyridine units, was characterized by FT-IR, (1)H and (13)C NMR, UV-visible and fluorescence spectroscopies. We investigated the relationship between changes of optical properties and protonation/deprotonation of PPyPE containing pyridine units in solution. Addition of HCl decreased and red-shifted the fluorescence intensity of the conjugated polymers that contained pyridine rings; fluorescence intensity of the polymers increased upon addition of NaOH solution. The synthesized PPyPE was found to be an effective and reusable chemical sensor for pH sensing. Copyright © 2015 John Wiley & Sons, Ltd.

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

Science.gov (United States)

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

2010-03-29

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

A KIND OF FLUORESCENCE PROBE TO STUDY THE KINETICS OF POLYMERIZATION PROCESS

Institute of Scientific and Technical Information of China (English)

YANG Guoqiang; WU Shikang

1994-01-01

Fluorescence properties of 1-phenyl-3-(4'-nitrophenyl) pyrazoline (PNP) were studied in bulk polymerization process of methylmethacrylate (MMA). The fluorescence intensity of PNP was enhanced and the emission maximum was blue shifted with the polymerization progress. In the period of auto-acceleration of the polymerization the enhancement of fluorescence intensity and blue shift of peak wavelength in spectra could be observed evidently. This means that the solvatochromic properties of PNP are influenced not only by the solvent polarity but also by the viscosity of the medium(especially by the phase transition). In solid state PNP emits from the charge transfer excited state without solvent relaxation. The transient emission spectra and the results from Bakhshiev model of solvent relaxation coincide with that from the polymerization experiment.

A novel fluorescent probe (dtpa-bis(cytosine)) for detection of Eu(III) in rare earth metal ions

Science.gov (United States)

Yang, Fan; Ren, Peipei; Liu, Guanhong; Song, Youtao; Bu, Naishun; Wang, Jun

2018-03-01

In this paper, a novel fluorescent probe, dtpa-bis(cytosine), was designed and synthesized for detecting europium (Eu3 +) ion. Upon addition of Eu3 + ions into the dtpa-bis(cytosine) solution, the fluorescence intensity can strongly be enhanced. Conversely, adding other rare earth metal ions, such as Y3 +, Ce3 +, Pr3 +, Nd3 +, Sm3 +, Gd3 +, Tb3 +, Dy3 +, Ho3 +, Er3 +, Yb3 + and Lu3 +, into dtpa-bis(cytosine) solution, the fluorescence intensity is decreased slightly. Some parameters affecting the fluorescence intensity of dtpa-bis(cytosine) solution in the presence of Eu3 + ions were investigated, including solution pH value, Eu3 + ion concentration and interfering substances. The detection mechanism of Eu3 + ion using dtpa-bis(cytosine) as fluorescent probe was proposed. Under optimum conditions, the fluorescence emission intensities of EuIII-dtpa-bis(cytosine) at 375 nm in the concentration range of 0.50 × 10- 5 mol • L- 1-5.00 × 10- 5 mol • L- 1 of Eu3 + ion display a better linear relationship. The limit of detection (LOD) was determined as 8.65 × 10- 7 mol • L- 1 and the corresponding correlation coefficient (R2) of the linear equation is 0.9807. It is wished that the proposed method could be applied for sensitively and selectively detecting Eu3 + ion.

Fluorescence blinking in MEH-PPV single molecules at low temperature

International Nuclear Information System (INIS)

Mirzov, O.; Cichos, F.; Borczyskowski, C. von; Scheblykin, I.

2005-01-01

Fluorescence intensity transients of single molecules of the conjugated polymer poly[2-methoxy,5-(2'-ethylhexyloxy)-p-phenylene-vinylene] (MEH-PPV) were studied at 15 K. Fluorescence blinking behavior was observed despite the expected low-temperature suppression of energy migration in such disordered molecular systems. Presence of the fluorescence blinking effect at 15 K indicates that the single molecules possess a collapsed conformation with characteristic size of not more than several nanometers, which corresponds to only a few exciton hops over a polymer chain

Application of the nuclear x-ray fluorescence method to prospecting for gold in-situ

International Nuclear Information System (INIS)

Zhang, Y.; Xie, T.; Zhou, S.; Ge, L.

1989-01-01

Arsenic and chalcophile elements are often associated with gold, and can be considered indicator elements when prospecting for gold deposits. The nuclear geophysics X-ray fluorescence method can be used to search for hidden gold deposits by measuring fluorescence intensities of the indicator elements in situ. The method can speed geologic investigation and reduce exploration cost. Three types of portable radioisotope X-ray fluorescence analyzers, designed and manufactured by Chengdu College of Geology and Chongqing Geological Instrument Factory, are briefly introduced. These analyzers are widely used in different stages of geologic investigation for gold in China. In the two case histories presented five anomalous zones of X-ray fluorescence intensity related to gold mineralization are located and one hidden gold deposit is discovered with gold content of 23 g/t

Changes to processes in estuaries and coastal waters due to intense multiple pressures - An introduction and synthesis

Science.gov (United States)

Mitchell, Steven B.; Jennerjahn, Tim C.; Vizzini, Salvatrice; Zhang, Weiguo

2015-04-01

From the 2013 ECSA conference 'Estuaries and Coastal Areas in Times of Intense Change' a theme emerged that has ended up being the focus of this Special Issue of Estuarine Coastal and Shelf Science, namely 'Changes to processes in estuaries and coastal waters due to intense multiple pressures'. Many parts of the world are continuing to experience unprecedented rates of economic growth, and those responsible for managing coastal and estuarine areas must respond accordingly. At the same time, global climate change and sea level rise are also continuing, placing new or more intense pressures on coastal areas that must be dealt with in ways that are as far as possible managed as a result of good scientific understanding. There are other pressures too, which depend on the system concerned. This article provides an overview of the papers contained within the Special Issue and provides a discussion of how these fit within the main theme of intense multiple stressors, considering how a balance can be achieved between the needs of various different stakeholders and interest groups, and the sustainability of the system concerned. We categorise the papers in four main groupings: (1) stressors related to sea level rise; (2) stressors related to changes in fresh water inputs; (3) stressors related to anthropogenic pollution; and (4) the use of indicators as a means of assessing the effects of stressors, and reflect on the fact that despite the diversity of different challenges and geographical regions involved many of the approaches and discussions contained within the Special Issue have strong similarities, leading to a set of overarching principles that should be considered when making recommendations on management strategies.

A chromenoquinoline-based fluorescent off-on thiol probe for bioimaging.

Science.gov (United States)

Kand, Dnyaneshwar; Kalle, Arunasree Marasanapalli; Varma, Sreejith Jayasree; Talukdar, Pinaki

2012-03-11

A new chromenoquinoline-based fluorescent off-on thiol probe 2 is reported. In aqueous buffer solutions at physiological pH, the probe exhibited 223-fold enhancement in fluorescence intensity by a Michael addition of cysteine to the maleimide appended to a chromenoquinoline. Cell permeability and live cell imaging of thiols are also demonstrated. This journal is © The Royal Society of Chemistry 2012

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

Science.gov (United States)

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

2009-01-01

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

ZnSe quantum dots based fluorescence quenching method for determination of paeoniflorin

International Nuclear Information System (INIS)

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

2014-01-01

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

ZnSe quantum dots based fluorescence quenching method for determination of paeoniflorin

Energy Technology Data Exchange (ETDEWEB)

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

2014-01-15

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

A red fluorescent nude mouse model of human endometriosis: advantages of a non-invasive imaging method.

Science.gov (United States)

Wang, Ningning; Hong, Shanshan; Tan, Jinfeng; Ke, Peiqi; Liang, Lili; Fei, Hui; Liu, Bin; Liu, Liqun; Liu, Yongdong; Yu, Bingjun

2014-05-01

To establish red fluorescent human endometriosis lesions in a nude mouse model and dynamically and non-invasively to compare intraperitoneal and subcutaneous injection models. Primary cultures of endometrial stromal cells (ESCs) and epithelial cells (EECs) isolated from 24 patients with a normal uterine cavity were transfected with 2.5×10(8) (Group 1) and 1.25×10(8) (Group 2) plaque-forming units (PFU) of adenovirus encoding red fluorescent protein (Ad-RFP). Transfection efficiencies, fluorescence intensity and apoptosis rate of the two types of cells were compared in vitro. A mixture of 2.5×10(8) PFU Ad-RFP-infected approximately 400 EECs cell mass and 2×10(6) ESCs for 36h was injected individually into 24 female nude mice subcutaneously (Group A) or intraperitoneally (Group B). From Day 5 after injection, an in vivo imaging system (IVIS) was used to non-invasively observe and compare the lesions of the two groups every week until Day 33. Specifically, the fluorescent intensity, positive rates, persistence time and lesion weight in the implanted human endometriosis lesions were compared. A parametric Student's t-test and two-way analysis of variance were used for statistical analysis. Compared with 1.25×10(8) PFU RFP, a titre of 2.5×10(8) PFU RFP ESCs and EECs incubated for 36h exhibited higher transfection efficiencies and higher fluorescence intensities in vitro. In vivo imaging of the fluorescent human endometriosis lesions originating from an RFP titre of 2.5×10(8) PFU showed that the intensity and lesion weight in Group A were significantly higher than in Group B. However, the two groups had the same RFP-positive rates and fluorescence persistence. The structure of each lesion was evaluated by immunohistochemistry to confirm its human endometrial origin. The red fluorescent human endometriosis model established by subcutaneously injecting 2.5×10(8) PFU RFP-transfected stromal cells and epithelial cells into nude mice had a higher fluorescent positive

Platform for efficient switching between multiple devices in the intensive care unit.

Science.gov (United States)

De Backere, F; Vanhove, T; Dejonghe, E; Feys, M; Herinckx, T; Vankelecom, J; Decruyenaere, J; De Turck, F

2015-01-01

This article is part of the Focus Theme of METHODS of Information in Medicine on "Managing Interoperability and Complexity in Health Systems". Handheld computers, such as tablets and smartphones, are becoming more and more accessible in the clinical care setting and in Intensive Care Units (ICUs). By making the most useful and appropriate data available on multiple devices and facilitate the switching between those devices, staff members can efficiently integrate them in their workflow, allowing for faster and more accurate decisions. This paper addresses the design of a platform for the efficient switching between multiple devices in the ICU. The key functionalities of the platform are the integration of the platform into the workflow of the medical staff and providing tailored and dynamic information at the point of care. The platform is designed based on a 3-tier architecture with a focus on extensibility, scalability and an optimal user experience. After identification to a device using Near Field Communication (NFC), the appropriate medical information will be shown on the selected device. The visualization of the data is adapted to the type of the device. A web-centric approach was used to enable extensibility and portability. A prototype of the platform was thoroughly evaluated. The scalability, performance and user experience were evaluated. Performance tests show that the response time of the system scales linearly with the amount of data. Measurements with up to 20 devices have shown no performance loss due to the concurrent use of multiple devices. The platform provides a scalable and responsive solution to enable the efficient switching between multiple devices. Due to the web-centric approach new devices can easily be integrated. The performance and scalability of the platform have been evaluated and it was shown that the response time and scalability of the platform was within an acceptable range.

Pathological diagnosis of bladder cancer by image analysis of hypericin induced fluorescence cystoscopic images

Science.gov (United States)

Kah, James C. Y.; Olivo, Malini C.; Lau, Weber K. O.; Sheppard, Colin J. R.

2005-08-01

Photodynamic diagnosis of bladder carcinoma based on hypericin fluorescence cystoscopy has shown to have a higher degree of sensitivity for the detection of flat bladder carcinoma compared to white light cystoscopy. The potential of the photosensitizer hypericin-induced fluorescence in performing non-invasive optical biopsy to grade bladder cancer in vivo using fluorescence cystoscopic image analysis without surgical resection for tissue biopsy is investigated in this study. The correlation between tissue fluorescence and histopathology of diseased tissue was explored and a diagnostic algorithm based on fluorescence image analysis was developed to classify the bladder cancer without surgical resection for tissue biopsy. Preliminary results suggest a correlation between tissue fluorescence and bladder cancer grade. By combining both the red-to-blue and red-to-green intensity ratios into a 2D scatter plot yields an average sensitivity and specificity of around 70% and 85% respectively for pathological cancer grading of the three different grades of bladder cancer. Therefore, the diagnostic algorithm based on colorimetric intensity ratio analysis of hypericin fluorescence cystoscopic images developed in this preliminary study shows promising potential to optically diagnose and grade bladder cancer in vivo.

Fluorescence quenching of Rhodamine B base by two amines

Science.gov (United States)

Bakkialakshmi, S.; Selvarani, P.; Chenthamarai, S.

2013-03-01

Fluorescence quenching of Rhodamine B base (RhB) in DMF solution has been studied at different concentrations of the amine Triethyl amine (TEA) and n-butyl amine (NBA) at room temperature. It has been observed that the fluorescence intensity of RhB decrease with increase in the concentration of the TEA and NBA. It has been observed that the quenching due to amines proceeds via dynamic quenching process. The rate constants for the quenching process have been calculated using Stern-Volmer equation. Time resolved fluorescence study and 1H NMR spectral study have also been carried out and discussed.

Photoacoustic-fluorescence in vitro flow cytometry for quantification of absorption, scattering and fluorescence properties of the cells

Science.gov (United States)

Nedosekin, D. A.; Sarimollaoglu, M.; Foster, S.; Galanzha, E. I.; Zharov, V. P.

2013-03-01

Fluorescence flow cytometry is a well-established analytical tool that provides quantification of multiple biological parameters of cells at molecular levels, including their functional states, morphology, composition, proliferation, and protein expression. However, only the fluorescence and scattering parameters of the cells or labels are available for detection. Cell pigmentation, presence of non-fluorescent dyes or nanoparticles cannot be reliably quantified. Herewith, we present a novel photoacoustic (PA) flow cytometry design for simple integration of absorbance measurements into schematics of conventional in vitro flow cytometers. The integrated system allow simultaneous measurements of light absorbance, scattering and of multicolor fluorescence from single cells in the flow at rates up to 2 m/s. We compared various combinations of excitation laser sources for multicolor detection, including simultaneous excitation of PA and fluorescence using a single 500 kHz pulsed nanosecond laser. Multichannel detection scheme allows simultaneous detection of up to 8 labels, including 4 fluorescent tags and 4 PA colors. In vitro PA-fluorescence flow cytometer was used for studies of nanoparticles uptake and for the analysis of cell line pigmentation, including genetically encoded melanin expression in breast cancer cell line. We demonstrate that this system can be used for direct nanotoxicity studies with simultaneous quantification of nanoparticles content and assessment of cell viability using a conventional fluorescent apoptosis assays.

Diagnostic imaging of cervical intraepithelial neoplasia based on hematoxylin and eosin fluorescence.

Science.gov (United States)

Castellanos, Mario R; Szerszen, Anita; Gundry, Stephen; Pirog, Edyta C; Maiman, Mitchell; Rajupet, Sritha; Gomez, John Paul; Davidov, Adi; Debata, Priya Ranjan; Banerjee, Probal; Fata, Jimmie E

2015-07-25

Pathological classification of cervical intraepithelial neoplasia (CIN) is problematic as it relies on subjective criteria. We developed an imaging method that uses spectroscopy to assess the fluorescent intensity of cervical biopsies derived directly from hematoxylin and eosin (H&E) stained tissues. Archived H&E slides were identified containing normal cervical tissue, CIN I, and CIN III cases, from a Community Hospital and an Academic Medical Center. Cases were obtained by consensus review of at least 2 senior pathologists. Images from H&E slides were captured first with bright field illumination and then with fluorescent illumination. We used a Zeiss Axio Observer Z1 microscope and an AxioVision 4.6.3-AP1 camera at excitation wavelength of 450-490 nm with emission captured at 515-565 nm. The 32-bit grayscale fluorescence images were used for image analysis. We reviewed 108 slides: 46 normal, 33 CIN I and 29 CIN III. Fluorescent intensity increased progressively in normal epithelial tissue as cells matured and advanced from the basal to superficial regions of the epithelium. In CIN I cases this change was less prominent as compared to normal. In high grade CIN lesions, there was a slight or no increase in fluorescent intensity. All groups examined were statistically different. Presently, there are no markers to help in classification of CIN I-III lesions. Our imaging method may complement standard H&E pathological review and provide objective criteria to support the CIN diagnosis.

Laser-induced fluorescence in the detection of esophageal carcinoma

Science.gov (United States)

Wang, Kenneth K.; Gutta, Kumar; Laukka, Mark A.; Densmore, John

1995-01-01

Laser induced fluorescence (LIF) is a technique which can perform an 'optical biopsy' of gastrointestinal mucosa. LIF was performed in resected specimens using a pulsed N2-laser coupled fiberoptically to a probe. Fluorescence was measured using a 0.2 meter spectroscope with an intensified photodiode array. Measurements were made on fresh (esophagus, and adenocarcinoma. Each tissue section was examined using an optical probe consisting of a central fiber for delivering the excitation energy and a 6 fiber bundle surrounding the central fiber for detection of the fluorescence. An excitation wavelength of 337 nm was used which generated 3-ns pulses while fluorescence intensities were acquired from 300-800 nm. Spectra were obtained from each section in a standardized fashion and background spectra subtracted. Fluorescence readings were taken from 54 normal esophageal sections and 32 sections of adenocarcinoma. A fluorescence index obtained from the tumor sections was 0.68+/- 0.01 compared with 0.51+/- 0.01 for the normal sections (pesophagus with good accuracy.

Hyper-spectral modulation fluorescent imaging using double acousto-optical tunable filter based on TeO2-crystals

International Nuclear Information System (INIS)

Zaytsev, Kirill I; Perchik, Alexey V; Chernomyrdin, Nikita V; Yurchenko, Stanislav O; Kudrin, Konstantin G; Reshetov, Igor V

2015-01-01

We have proposed a method for hyper-spectral fluorescent imaging based on acousto-optical filtering. The object of interest was pumped using ultraviolet radiation of mercury lamp equipped with monochromatic excitation filter with the window of transparency centered at 365 nm. Double TeO 2 -based acousto-optical filter, tunable in range from 430 to 780 nm and having 2 nm bandwidth of spectral transparency, was used in order to detect quasimonochromatic images of object fluorescence. Modulating of ultraviolet pump intensity was used in order to reduce an impact of non-fluorescent background on the sample fluorescent imaging. The technique for signal-to-noise ratio improvement, based on fluorescence intensity estimation via digital processing of modulated video sequence of fluorescent object, was introduced. We have implemented the proposed technique for the test sample studying and we have discussed its possible applications

Impurity monitoring by laser-induced fluorescence techniques

International Nuclear Information System (INIS)

Gelbwachs, J.A.

1984-01-01

Laser-induced fluorescence spectroscopy can provide a highly sensitive and selective means of detecting atomic and ionic impurities. Because the photodetector can be physically isolated from the laser-excited region, these techniques can be applied to monitoring in hostile environments. The basic concepts behind fluorescence detection are reviewed. Saturated optical excitation is shown to maximize impurity atom emission yield while mitigating effects of laser intensity fluctuations upon absolute density calibration. Monitoring in high- and low-pressure monitoring environments is compared. Methods to improve detection sensitivity by luminescence background suppression are presented

Three-dimensional mapping of fluorescent nanoparticles using incoherent digital holography.

Science.gov (United States)

Yanagawa, Takumi; Abe, Ryosuke; Hayasaki, Yoshio

2015-07-15

Three-dimensional mapping of fluorescent nanoparticles was performed by using incoherent digital holography. The positions of the nanoparticles were quantitatively determined by using Gaussian fitting of the axial- and lateral-diffraction distributions through position calibration from the observation space to the sample space. It was found that the axial magnification was constant whereas the lateral magnification linearly depended on the axial position of the fluorescent nanoparticles. The mapping of multiple fluorescent nanoparticles fixed in gelatin and a single fluorescent nanoparticle manipulated with optical tweezers in water were demonstrated.

Red and Green Fluorescence from Oral Biofilms.

Science.gov (United States)

Volgenant, Catherine M C; Hoogenkamp, Michel A; Krom, Bastiaan P; Janus, Marleen M; Ten Cate, Jacob M; de Soet, Johannes J; Crielaard, Wim; van der Veen, Monique H

2016-01-01

Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation) as compared to the sucrose grown biofilms (cariogenic simulation). Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red) were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

Red and Green Fluorescence from Oral Biofilms.

Directory of Open Access Journals (Sweden)

Catherine M C Volgenant

Full Text Available Red and green autofluorescence have been observed from dental plaque after excitation by blue light. It has been suggested that this red fluorescence is related to caries and the cariogenic potential of dental plaque. Recently, it was suggested that red fluorescence may be related to gingivitis. Little is known about green fluorescence from biofilms. Therefore, we assessed the dynamics of red and green fluorescence in real-time during biofilm formation. In addition, the fluorescence patterns of biofilm formed from saliva of eight different donors are described under simulated gingivitis and caries conditions. Biofilm formation was analysed for 12 hours under flow conditions in a microfluidic BioFlux flow system with high performance microscopy using a camera to allow live cell imaging. For fluorescence images dedicated excitation and emission filters were used. Both green and red fluorescence were linearly related with the total biomass of the biofilms. All biofilms displayed to some extent green and red fluorescence, with higher red and green fluorescence intensities from biofilms grown in the presence of serum (gingivitis simulation as compared to the sucrose grown biofilms (cariogenic simulation. Remarkably, cocci with long chain lengths, presumably streptococci, were observed in the biofilms. Green and red fluorescence were not found homogeneously distributed within the biofilms: highly fluorescent spots (both green and red were visible throughout the biomass. An increase in red fluorescence from the in vitro biofilms appeared to be related to the clinical inflammatory response of the respective saliva donors, which was previously assessed during an in vivo period of performing no-oral hygiene. The BioFlux model proved to be a reliable model to assess biofilm fluorescence. With this model, a prediction can be made whether a patient will be prone to the development of gingivitis or caries.

Laser scanning endoscope via an imaging fiber bundle for fluorescence imaging

Science.gov (United States)

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

1994-12-01

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

Fluorescence properties of Schiff base - N,N‧-bis(salicylidene) - 1,2-Phenylenediamine in presence of bile acid host

Science.gov (United States)

Roy, Nayan; Paul, Pradip C.; Singh, T. Sanjoy

2015-05-01

Fluorescence properties of Schiff base - N,N‧-bis(salicylidene) - 1,2-phenylenediamine (LH2) is used to study the micelles formed by aggregation of different important bile acids like cholic acid, deoxycholic acid, chenodeoxycholic acid and glycocholic acid by steady state and picosecond time-resolved fluorescence spectroscopy. The fluorescence band intensity was found out to increase with concomitant red shift with gradual addition of different bile acids. Binding constant of the probe with different bile acids as well as critical micelle concentration was obtained from the variation of fluorescence intensity on increasing concentration of bile acids in the medium. The increase in fluorescence quantum yields, fluorescence decay times and substantial decrease in nonradiative decay rate constants in bile acids micellar environment points to the restricted motion of the fluorophore inside the micellar subdomains.

New dual emission fluorescent sensor for pH and Pb(II) based on bis(napfthalimide) derivative

Energy Technology Data Exchange (ETDEWEB)

Pina-Luis, Georgina, E-mail: gpinaluis@yahoo.com [Centro de Graduados e Investigacion en Quimica, Instituto Tecnologico de Tijuana, AP 1166, Tijuana 22500, BC (Mexico); Martinez-Quiroz, Marisela; Ochoa-Teran, Adrian [Centro de Graduados e Investigacion en Quimica, Instituto Tecnologico de Tijuana, AP 1166, Tijuana 22500, BC (Mexico); Santacruz-Ortega, Hisila [Departamento de investigacion en Polimeros y Materiales, Universidad de Sonora, Hermosillo, Sonora 83000 (Mexico); Mendez-Valenzuela, Eduardo [Centro de Graduados e Investigacion en Quimica, Instituto Tecnologico de Tijuana, AP 1166, Tijuana 22500, BC (Mexico)

2013-02-15

This paper describes a novel dual emission bis-1,8-naphthalimide sensor for selective determination of pH and Pb{sup 2+} ions. The influence of the variability in the backbone that links the two fluorophores (naphthalimides) as a function of pH and metal ions was studied by UV-visible and fluorescence spectroscopy. Compounds 1(a-d) with different length alkyl linkers (CH{sub 2}){sub n} (n=1, 2, 4 and 6) showed no excimer formation in aqueous solution. Fluorescence emission of these derivatives varied in a narrow range of pH (5-8) and was only slightly influenced by the addition of metal ions in CH{sub 3}CN solutions. However, derivative 1e with amino-containing spacer (CH{sub 2}-NH-CH{sub 2}) showed excimer emission in aqueous solution, a wide response to pH (2.5-9.5) and fluorescence enhancement with selective behavior towards metal ions. The pH sensor based in derivative 1e has a sufficient selectivity for practical pH monitoring in the presence of Li{sup +}, Na{sup +}, K{sup +}, Cs{sup +}, Ca{sup 2+}, Mg{sup 2+}, Ba{sup 2+}, Cu{sup 2+}, Pb{sup 2+}, Ni{sup 2+}, Zn{sup 2+} and Cd{sup 2+}. The coordination chemistry of these complexes was studied by UV-Vis, fluorescence and {sup 1}H NMR. This chemosensor displayed high selectivity fluorescence enhancement toward Pb{sup 2+} ions in the presence of the metals ions mentioned in CH{sub 3}CN solutions. Competitive assays show that a 1-fold of metal cations in each case, compared with Pb{sup 2+} ions, results in less than {+-}5% fluorescence intensity changes. Linear calibration up to 1 Multiplication-Sign 10{sup -5} M for Pb(II) ions (R=0.9968) was obtained and detection limit resulted of 5.0 Multiplication-Sign 10{sup -8} M. - Highlights: Black-Right-Pointing-Pointer A novel dual emission bis-1,8-naphthalimide sensor for pH and Pb{sup 2+} ions is synthetized. Black-Right-Pointing-Pointer The excimer formation depends on the spacer that links the two naphthalimide groups. Black-Right-Pointing-Pointer Bis

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

International Nuclear Information System (INIS)

Bian Weiwei; Wang Yusheng; Zhu Xiaojing; Jiang Chongqiu

2006-01-01

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

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

Science.gov (United States)

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.

Fluorescent analysis of interaction of flavonols with hemoglobin and bovine serum albumin

Science.gov (United States)

Sentchouk, V. V.; Bondaryuk, E. V.

2007-09-01

We have studied the fluorescent properties of flavonols (quercetin, fisetin, morin, rutin) with the aim of studying possible interaction with hemoglobin and bovine serum albumin (BSA). We observed an increase in the intensity of intrinsic fluorescence for all the flavonols except rutin in the presence of BSA. From the changes in the fluorescence spectra, we concluded that tautomeric forms are formed on interaction with hemoglobin. We determined the interconnection between the structure of related flavonols and their fluorescent properties on interaction with proteins, and we determined the binding constants for binding with BSA and hemoglobin.

Noise analysis of a white-light supercontinuum light source for multiple wavelength confocal laser scanning fluorescence microscopy

Energy Technology Data Exchange (ETDEWEB)

McConnell, Gail [Centre for Biophotonics, Strathclyde Institute for Biomedical Sciences, University of Strathclyde, 27 Taylor Street, Glasgow, G4 0NR (United Kingdom)

2005-08-07

Intensity correlations of a Ti : sapphire, Kr/Ar and a white-light supercontinuum were performed to quantify the typical signal amplitude fluctuations and hence ascertain the comparative output stability of the white-light supercontinuum source for confocal laser scanning microscopy (CLSM). Intensity correlations across a two-pixel sample (n = 1000) of up to 98%, 95% and 94% were measured for the Ti : sapphire, Kr/Ar and white-light supercontinuum source, respectively. The white-light supercontinuum noise level is therefore acceptable for CLSM, with the added advantage of wider wavelength flexibility over traditional CLSM excitation sources. The relatively low-noise white-light supercontinuum was then used to perform multiple wavelength sequential CLSM of guinea pig detrusor to confirm the reliability of the system and to demonstrate system flexibility.

Novel lanthanide pH fluorescent probes based on multiple emissions and its visible-light-sensitized feature

Energy Technology Data Exchange (ETDEWEB)

Lin, Jintai [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Zheng, Yuhui, E-mail: yhzheng78@scnu.edu.cn [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Wang, Qianming, E-mail: qmwang@scnu.edu.cn [Key Laboratory of Theoretical Chemistry of Environment, Ministry of Education, School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Guangzhou Key Laboratory of Materials for Energy Conversion and Storage, Guangzhou 510006 (China); State Key Laboratory of Chemo/Biosensing and Chemometrics, Hunan University, Changsha 410082 (China); Zeng, Zhi [School of Chemistry and Environment, South China Normal University, Guangzhou 510006 (China); Zhang, Cheng Cheng [Departments of Physiology and Developmental Biology, University of Texas Southwestern Medical Center, Dallas (United States)

2014-08-11

Graphical abstract: A new type of Eu(III) ofloxacin complex as the fluorescent pH indicator has been reported. Compared to pure ligand, the complex offers more distinguished color changes (green–red–blue) derived from both lanthanide line emissions and the secondary ionization steps of ofloxacin. - Highlights: • The pH probe offers a very wide working range in water (pH 1–14). • The emission changes have multiple colors. • Long-lived excited state lifetimes of Eu(III) has been used. • Two types of pH sensitive hydrogels were fabricated. - Abstract: A new type of Eu(III) ofloxacin complex as the fluorescent pH indicator has been presented. Compared to pure ligand, the complex offers more distinguished color changes (green–red–blue) derived from both lanthanide line emissions and the secondary ionization steps of ofloxacin. During the concentration dependence experiments, the photoluminescence studies on the complex showed that the excitation of this pH probe can occur at a very long wavelength which extends to visible range (Ex = 427 nm). Furthermore, the functional complex was successfully incorporated into soft networks and two novel luminescent hydrogels (rod and film) were fabricated. The soft materials also exhibited specific responses towards the pH variation. Finally, the onion cell-stain experiments were carried out to further confirm the validity of pH dependence and the results support the idea that the material will be suitable for monitoring biological samples in the future.

Plasmonics Enhanced Smartphone Fluorescence Microscopy

KAUST Repository

Wei, Qingshan

2017-05-12

Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.

Plasmonics Enhanced Smartphone Fluorescence Microscopy

KAUST Repository

Wei, Qingshan; Acuna, Guillermo; Kim, Seungkyeum; Vietz, Carolin; Tseng, Derek; Chae, Jongjae; Shir, Daniel; Luo, Wei; Tinnefeld, Philip; Ozcan, Aydogan

2017-01-01

Smartphone fluorescence microscopy has various applications in point-of-care (POC) testing and diagnostics, ranging from e.g., quantification of immunoassays, detection of microorganisms, to sensing of viruses. An important need in smartphone-based microscopy and sensing techniques is to improve the detection sensitivity to enable quantification of extremely low concentrations of target molecules. Here, we demonstrate a general strategy to enhance the detection sensitivity of a smartphone-based fluorescence microscope by using surface-enhanced fluorescence (SEF) created by a thin metal-film. In this plasmonic design, the samples are placed on a silver-coated glass slide with a thin spacer, and excited by a laser-diode from the backside through a glass hemisphere, generating surface plasmon polaritons. We optimized this mobile SEF system by tuning the metal-film thickness, spacer distance, excitation angle and polarization, and achieved ~10-fold enhancement in fluorescence intensity compared to a bare glass substrate, which enabled us to image single fluorescent particles as small as 50 nm in diameter and single quantum-dots. Furthermore, we quantified the detection limit of this platform by using DNA origami-based brightness standards, demonstrating that ~80 fluorophores per diffraction-limited spot can be readily detected by our mobile microscope, which opens up new opportunities for POC diagnostics and sensing applications in resource-limited-settings.

Spatial distribution of fluorescent light emitted from neon and nitrogen excited by low energy electron beams

International Nuclear Information System (INIS)

Morozov, A.; Kruecken, R.; Ulrich, A.; Wieser, J.

2006-01-01

Side-view intensity profiles of fluorescent light were measured for neon and nitrogen excited with 12 keV electron beams at gas pressures from 250 to 1400 hPa. The intensity profiles were compared with theoretical profiles calculated using the CASINO program which performs Monte Carlo simulations of electron scattering. It was assumed that the spatial distribution of fluorescent intensity is directly proportional to the spatial distribution of energy loss by primary electrons. The comparison shows good correlation of experimental data and the results of numeric simulations

Fluorescein Derivatives in Intravital Fluorescence Imaging

Directory of Open Access Journals (Sweden)

Michael S. Roberts

2013-08-01

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

Subcellular localization of low-abundance human immunodeficiency virus nucleic acid sequences visualized by fluorescence in situ hybridization

International Nuclear Information System (INIS)

Lawrence, J.B.; Marselle, L.M.; Byron, K.S.; Johnson, C.V.; Sullivan, J.L.; Singer, R.H.

1990-01-01

Detection and subcellular localization of human immunodeficiency virus (HIV) were investigated using sensitive high-resolution in situ hybridization methodology. Lymphocytes infected with HIV in vitro or in vivo were detected by fluorescence after hybridization with either biotin or digoxigenin-labeled probes. At 12 hr after infection in vitro, a single intense signal appeared in the nuclei of individual cells. Later in infection, when cytoplasmic fluorescence became intense, multiple nuclear foci frequently appeared. The nuclear focus consisted of newly synthesized HIV RNA as shown by hybridization in the absence of denaturation and by susceptibility to RNase and actinomycin D. Virus was detected in patient lymphocytes and it was shown that a singular nuclear focus also characterizes cells infected in vivo. The cell line 8E5/LAV containing one defective integrated provirus revealed a similar focus of nuclear RNA, and the single integrated HIV genome was unequivocally visualized on a D-group chromosome. This demonstrates an extremely sensitive single-cell assay for the presence of a single site of HIV transcription in vitro and in vivo and suggests that it derives from one (or very few) viral genomes per cell. In contrast, productive Epstein-Barr virus infection exhibited many foci of nuclear RNA per cell

Light emission from compound eye with conformal fluorescent coating

Science.gov (United States)

Martín-Palma, Raúl J.; Miller, Amy E.; Pulsifer, Drew P.; Lakhtakia, Akhlesh

2015-03-01

Compound eyes of insects are attractive biological systems for engineered biomimicry as artificial sources of light, given their characteristic wide angular field of view. A blowfly eye was coated with a thin conformal fluorescent film, with the aim of achieving wide field-of-view emission. Experimental results showed that the coated eye emitted visible light and that the intensity showed a weaker angular dependence than a fluorescent thin film deposited on a flat surface.

The impact of fluorescent dyes on the performances of polystyrene-based plastic scintillators

International Nuclear Information System (INIS)

Zhu, Jun; Deng, Cheng; Jiang, Huimin; Zheng, Zhanlong; Gong, Rui; Bi, Yutie; Zhang, Lin; Lin, Runxiong

2016-01-01

To investigate the influence of both the first luminescent additive and the wavelength-shifter on the performance of plastic scintillator, a series of polystyrene-based scintillator had been prepared by thermal polymerization. Three first luminescent additives (PPO, p-TP and b-PBD) and four wavelength-shifters (POPOP, Bis-MSB, Me-MSB and DPA) were added to the scintillators respectively. The comparison results showed that PPO and POPOP were the most adequate fluorescent dyes for the polystyrene-based plastic scintillator. Moreover, with the increase of the concentration of PPO and POPOP, the fluorescence intensity and light yield were increased firstly and then decreased. The plastic scintillator containing 2% PPO and 0.02% POPOP had the highest fluorescence intensity and light yield.

The impact of fluorescent dyes on the performances of polystyrene-based plastic scintillators

Energy Technology Data Exchange (ETDEWEB)

Zhu, Jun [Research Center of Laser Fusion, China Academy of Engineering Physics, Mianyang 621900 (China); Engineering Research Center of High Performance Polymer and Molding Technology, Ministry of Education, Qingdao University of Science and Technology, Qingdao 266042 (China); Joint Laboratory for Extreme Conditions Matter Properties, Southwest University of Science and Technology and Research Center of Laser Fusion, China Academy of Engineering Physics, Mianyang 621010 (China); Deng, Cheng; Jiang, Huimin [Engineering Research Center of High Performance Polymer and Molding Technology, Ministry of Education, Qingdao University of Science and Technology, Qingdao 266042 (China); Zheng, Zhanlong; Gong, Rui [Research Center of Laser Fusion, China Academy of Engineering Physics, Mianyang 621900 (China); Joint Laboratory for Extreme Conditions Matter Properties, Southwest University of Science and Technology and Research Center of Laser Fusion, China Academy of Engineering Physics, Mianyang 621010 (China); Bi, Yutie [Joint Laboratory for Extreme Conditions Matter Properties, Southwest University of Science and Technology and Research Center of Laser Fusion, China Academy of Engineering Physics, Mianyang 621010 (China); Zhang, Lin, E-mail: zhlmy@sina.com [Research Center of Laser Fusion, China Academy of Engineering Physics, Mianyang 621900 (China); Lin, Runxiong, E-mail: qdlrx@qust.edu.cn [Engineering Research Center of High Performance Polymer and Molding Technology, Ministry of Education, Qingdao University of Science and Technology, Qingdao 266042 (China)

2016-11-01

To investigate the influence of both the first luminescent additive and the wavelength-shifter on the performance of plastic scintillator, a series of polystyrene-based scintillator had been prepared by thermal polymerization. Three first luminescent additives (PPO, p-TP and b-PBD) and four wavelength-shifters (POPOP, Bis-MSB, Me-MSB and DPA) were added to the scintillators respectively. The comparison results showed that PPO and POPOP were the most adequate fluorescent dyes for the polystyrene-based plastic scintillator. Moreover, with the increase of the concentration of PPO and POPOP, the fluorescence intensity and light yield were increased firstly and then decreased. The plastic scintillator containing 2% PPO and 0.02% POPOP had the highest fluorescence intensity and light yield.

Combining electrophoresis with detection under ultraviolet light and multiple ultrafiltration for isolation of humic fluorescence fractions.

Science.gov (United States)

Trubetskaya, Olga E; Shaloiko, Lubov A; Demin, Dmitrii V; Marchenkov, Victor V; Proskuryakov, Ivan I; Coelho, Christian; Trubetskoj, Oleg A

2011-04-01

Polyacrylamide gel electrophoresis of chernozem soil humic acids (HAs) followed by observation under UV (312 nm) excitation light reveals new low molecular weight (MW) fluorescent fractions. Ultrafiltration of HAs sample in 7 M urea on a membrane of low nominal MW retention (NMWR, 5 kDa) was repetitively used for separation of fluorescent and non-fluorescent species. Thirty ultrafiltrates and the final retentate R were obtained. Fluorescence maxima of separate ultrafiltrates were different and non-monotonously changed in the range of 475-505 nm. Fluorescence maxima of less than 490 nm were detected only in the four first utrafiltrates. For further physical-chemical analyses all utrafiltrates were combined into a fraction called UFchernozem soil HAs complex. Copyright © 2011 Elsevier B.V. All rights reserved.

Computer generated holography with intensity-graded patterns

Directory of Open Access Journals (Sweden)

Rossella Conti

2016-10-01

Full Text Available Computer Generated Holography achieves patterned illumination at the sample plane through phase modulation of the laser beam at the objective back aperture. This is obtained by using liquid crystal-based spatial light modulators (LC-SLMs, which modulate the spatial phase of the incident laser beam. A variety of algorithms are employed to calculate the phase modulation masks addressed to the LC-SLM. These algorithms range from simple gratings-and-lenses to generate multiple diffraction-limited spots, to iterative Fourier-transform algorithms capable of generating arbitrary illumination shapes perfectly tailored on the base of the target contour. Applications for holographic light patterning include multi-trap optical tweezers, patterned voltage imaging and optical control of neuronal excitation using uncaging or optogenetics. These past implementations of computer generated holography used binary input profile to generate binary light distribution at the sample plane. Here we demonstrate that using graded input sources, enables generating intensity graded light patterns and extend the range of application of holographic light illumination. At first, we use intensity-graded holograms to compensate for LC-SLM position dependent diffraction efficiency or sample fluorescence inhomogeneity. Finally we show that intensity-graded holography can be used to equalize photo evoked currents from cells expressing different level of chanelrhodopsin2 (ChR2, one of the most commonly used optogenetics light gated channels, taking into account the non-linear dependence of channel opening on incident light.

Synthesis of strongly fluorescent carbon quantum dots modified with polyamidoamine and a triethoxysilane as quenchable fluorescent probes for mercury(II)

International Nuclear Information System (INIS)

Tang, Wenjie; Wang, Yan; Wang, Panpan; Di, Junwei; Wu, Ying; Yang, Jianping

2016-01-01

This article reports on the synthesis of water dispersible carbon quantum dots (CDs) by a one-step hydrothermal method using polyamidoamine (PAMAM) and (3-aminopropyl)triethoxysilane (APTES) as a platform and passivant. The resulting CDs are highly uniform and finely dispersed. The synergistic effect between PAMAM and APTES on the surface of the CDs results in a fluorescence that is much brighter than that of CDs modified with either APTES or PAMAM only. The fluorescence of the co-modified CDs is quenched by Hg(II) ions at fairly low concentrations. Under the optimum conditions, the intensity of quenched fluorescence drops with Hg(II) concentration in the range from 0.2 nM to 10 μM, and the detection limit is 87 fM. The effect of potentially interfering cations on the fluorescence revealed a high selectivity for Hg 2+ . The fluorescent probe was applied to the determination of Hg(II) in (spiked) waters and milk and gave recoveries between 95.6 and 107 %, with relative standard deviation between 4.4 and 6.0 %. (author)

The fluorescence properties of aerosol larger than 0.8 μm in urban and tropical rainforest locations

Directory of Open Access Journals (Sweden)

A. M. Gabey

2011-06-01

Full Text Available UV-LIF measurements were performed on ambient aerosol in Manchester, UK (urban city centre, winter and Borneo, Malaysia (remote, tropical using a Wide Issue Bioaerosol Spectrometer, version 3 (WIBS3. These sites are taken to represent environments with minor and significant primary biological aerosol (PBA influences respectively, and the urban dataset describes the fluorescent background aerosol against which PBA must be identified by researchers using LIF. The ensemble aerosol at both sites was characterised over 2–3 weeks by measuring the fluorescence intensity and optical equivalent diameter (D_P of single particles sized 0.8 ≤ D_P ≤ 20 μm. Filter samples were also collected for a subset of the Manchester campaign and analysed using energy dispersive X-Ray (EDX spectroscopy and environmental scanning electron microscopy (ESEM, which revealed mostly non-PBA at D ≤ 1 μm.

The WIBS3 features three fluorescence channels: the emission following a 280 nm excitation is recorded at 310–400 nm (channel F1 and 400–600 nm (F2, and fluorescence excited at 350 nm is detected at 400–600 nm (F3. In Manchester the primary size mode of fluorescent and non-fluorescent material was present at 0.8–1.2 μm, with a secondary fluorescent mode at 2–4 μm. In Borneo non-fluorescent material peaked at 0.8–1.2 μm and fluorescent at 3–4 μm. Agreement between fluorescent number concentrations in each channel differed at the two sites, with F1 and F3 reporting similar concentrations in Borneo but F3 outnumbering F1 by a factor of 2–3 across the size spectrum in Manchester.

The fluorescence intensity in each channel generally rose with D_P at both sites with the exception of F1 intensity in Manchester, which peaked at D_P = 4 μm, causing a divergence between F1 and F3 intensity at larger D_P. This divergence and the differing

Intensity dependent absorption bleaching of high subband excitons in GaAs/AlGaAs multiple quantum wells

CERN Document Server

Shin, S H; Lee, E H; Chae, K M; Park, S H; Kim, U

1998-01-01

We have investigated the influence of carrier generation on the absorption bleaching of the n=2 and n=3 excitons in GaAs/AlGaAs multiple quantum wells (MQWs). With the excitation near the resonance of the n=1 exciton absorption, the long range coulomb screening and collision broadening had significant effects on the exciton bleaching. At low excitation intensity, the absorption bleaching of the n=2 exciton in 75 A-thick MQWs and that of the n=3 exciton in 150 A-thick MQWs were due to linewidth broadening by the collision broadening effect only. At high excitation intensity, however, the reduction of oscillator strength due to the long range coulomb screening contributed dominantly to absorption bleaching.

Predictors of intensive care unit refusal in French intensive care units: a multiple-center study.

Science.gov (United States)

Garrouste-Orgeas, Maité; Montuclard, Luc; Timsit, Jean-François; Reignier, Jean; Desmettre, Thibault; Karoubi, Philippe; Moreau, Delphine; Montesino, Laurent; Duguet, Alexandre; Boussat, Sandrine; Ede, Christophe; Monseau, Yannick; Paule, Thierry; Misset, Benoit; Carlet, Jean

2005-04-01

To identify factors associated with granting or refusing intensive care unit (ICU) admission, to analyze ICU characteristics and triage decisions, and to describe mortality in admitted and refused patients. Observational, prospective, multiple-center study. Four university hospitals and seven primary-care hospitals in France. None. Age, underlying diseases (McCabe score and Knaus class), dependency, hospital mortality, and ICU characteristics were recorded. The crude ICU refusal rate was 23.8% (137/574), with variations from 7.1% to 63.1%. The reasons for refusal were too well to benefit (76/137, 55.4%), too sick to benefit (51/137, 37.2%), unit too busy (9/137, 6.5%), and refusal by the family (1/137). In logistic regression analyses, two patient-related factors were associated with ICU refusal: dependency (odds ratio [OR], 14.20; 95% confidence interval [CI], 5.27-38.25; p refused patients, and 1.03 (95% CI, 0.28-1.75) for later-admitted patients. ICU refusal rates varied greatly across ICUs and were dependent on both patient and organizational factors. Efforts to define ethically optimal ICU admission policies might lead to greater homogeneity in refusal rates, although case-mix variations would be expected to leave an irreducible amount of variation across ICUs.

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

International Nuclear Information System (INIS)

Yu Fengshan; Li Lin; Chen Fang

2008-01-01

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

Assessing the Effectiveness of a Far-Red Fluorescent Reporter for Tracking Stem Cells In Vivo

Directory of Open Access Journals (Sweden)

Jing Zhou

2017-12-01

Full Text Available Far-red fluorescent reporter genes can be used for tracking cells non-invasively in vivo using fluorescence imaging. Here, we investigate the effectiveness of the far-red fluorescent protein, E2-Crimson (E2C, for tracking mouse embryonic cells (mESCs in vivo following subcutaneous administration into mice. Using a knock-in strategy, we introduced E2C into the Rosa26 locus of an E14-Bra-GFP mESC line, and after confirming that the E2C had no obvious effect on the phenotype of the mESCs, we injected them into mice and imaged them over nine days. The results showed that fluorescence intensity was weak, and cells could only be detected when injected at high densities. Furthermore, intensity peaked on day 4 and then started to decrease, despite the fact that tumour volume continued to increase beyond day 4. Histopathological analysis showed that although E2C fluorescence could barely be detected in vivo at day 9, analysis of frozen sections indicated that all mESCs within the tumours continued to express E2C. We hypothesise that the decrease in fluorescence intensity in vivo was probably due to the fact that the mESC tumours became more vascular with time, thus leading to increased absorbance of E2C fluorescence by haemoglobin. We conclude that the E2C reporter has limited use for tracking cells in vivo, at least when introduced as a single copy into the Rosa26 locus.

Fluorescent aggregates in naphthalene containing poly(urethane-urea)s

International Nuclear Information System (INIS)

Simas, E.R.; Akcelrud, Leni

2003-01-01

A series of segmented poly(urethane-urea)s containing naphthalene in the hard block and hexamethylene spacers in the soft block was prepared. The hard to soft segment ratio was varied systematically, to afford a series of polymers with various chromophore concentrations and a constant length of the chromophoric block, using a three-step synthetic procedure. The absorption, fluorescence and fluorescence-excitation spectra of solutions and films of the block copolymers provide strong evidence for aggregation. A red-shifted fluorescence spectrum peaking at 420 nm gains in intensity as the naphthalene concentration is increased. The excitation spectrum of this new emission is well to the red of the normal naphthalene absorption spectrum, consistent with the UV spectrum. Formation of a fluorescent ground state dimer (or higher aggregate) is proposed to account for these observations

Contribution of inner shell Compton ionization to the X-ray fluorescence line intensity

Science.gov (United States)

Fernández, Jorge E.; Scot, Viviana; Di Giulio, Eugenio

2016-10-01

The Compton effect is a potential ionization mechanism of atoms. It produces vacancies in inner shells that are filled with the same mechanism of atomic relaxation as the one following photo-absorption. This contribution to X-ray fluorescence emission is frequently neglected because the total Compton cross-section is apparently much lower than the photoelectric one at useful X-ray energies. However, a more careful analysis suggests that is necessary to consider single shell cross sections (instead of total cross sections) as a function of energy. In this article these Compton cross sections are computed for the shells K, L1-L3 and M1-M5 in the framework of the impulse approximation. By comparing the Compton and the photoelectric cross-section for each shell it is then possible to determine the extent of the Compton correction to the intensity of the corresponding characteristic lines. It is shown that for the K shell the correction becomes relevant for excitation energies which are too high to be influent in X-ray spectrometry. In contrast, for L and M shells the Compton contribution is relevant for medium-Z elements and medium energies. To illustrate the different grades of relevance of the correction, for each ionized shell, the energies for which the Compton contribution reaches the extent levels of 1, 5, 10, 20, 50 and 100% of the photoelectric one are determined for all the elements with Z = 11-92. For practical applications it is provided a simple formula and fitting coefficients to compute average correction levels for the shells considered.

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

Science.gov (United States)

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

2013-02-01

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

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

Science.gov (United States)

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

2014-04-03

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

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

Science.gov (United States)

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

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

[Ph-Sensor Properties of a Fluorescent Protein from Dendronephthya sp].

Science.gov (United States)

Pakhomov, A A; Chertkova, R V; Martynov, V I

2015-01-01

Genetically encoded biosensors based on fluorescent proteins are now widely applicable for monitoring pH changes in live cells. Here, we have shown that a fluorescent protein from Dendronephthya sp. (DendFP) exhibits a pronounced pH-sensitivity. Unlike most of known genetically encoded pH-sensors, fluorescence of the protein is not quenched upon medium acidification, but is shifting from the red to green spectral range. Therefore, quantitative measurements of intracellular pH are feasible by ratiometric comparison of emission intensities in the red and green spectral ranges, which makes DendFP advantageous compared with other genetically encoded pH-sensors.

Near-Membrane Refractometry Using Supercritical Angle Fluorescence.

Science.gov (United States)

Brunstein, Maia; Roy, Lopamudra; Oheim, Martin

2017-05-09

Total internal reflection fluorescence (TIRF) microscopy and its variants are key technologies for visualizing the dynamics of single molecules or organelles in live cells. Yet truly quantitative TIRF remains problematic. One unknown hampering the interpretation of evanescent-wave excited fluorescence intensities is the undetermined cell refractive index (RI). Here, we use a combination of TIRF excitation and supercritical angle fluorescence emission detection to directly measure the average RI in the "footprint" region of the cell during image acquisition. Our RI measurement is based on the determination on a back-focal plane image of the critical angle separating evanescent and far-field fluorescence emission components. We validate our method by imaging mouse embryonic fibroblasts and BON cells. By targeting various dyes and fluorescent-protein chimeras to vesicles, the plasma membrane, as well as mitochondria and the endoplasmic reticulum, we demonstrate local RI measurements with subcellular resolution on a standard TIRF microscope, with a removable Bertrand lens as the only modification. Our technique has important applications for imaging axial vesicle dynamics and the mitochondrial energy state or detecting metabolically more active cancer cells. Copyright © 2017. Published by Elsevier Inc.

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

International Nuclear Information System (INIS)

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

2016-01-01

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

Spirally-patterned pinhole arrays for long-term fluorescence cell imaging.

Science.gov (United States)

Koo, Bon Ung; Kang, YooNa; Moon, SangJun; Lee, Won Gu

2015-11-07

Fluorescence cell imaging using a fluorescence microscope is an extensively used technique to examine the cell nucleus, internal structures, and other cellular molecules with fluorescence response time and intensity. However, it is difficult to perform high resolution cell imaging for a long period of time with this technique due to necrosis and apoptosis depending on the type and subcellular location of the damage caused by phototoxicity. A large number of studies have been performed to resolve this problem, but researchers have struggled to meet the challenge between cellular viability and image resolution. In this study, we employ a specially designed disc to reduce cell damage by controlling total fluorescence exposure time without deterioration of the image resolution. This approach has many advantages such as, the apparatus is simple, cost-effective, and easily integrated into the optical pathway through a conventional fluorescence microscope.

An aptamer-based fluorescence bio-sensor for chiral recognition of arginine enantiomers.

Science.gov (United States)

Yuan, Haiyan; Huang, Yunmei; Yang, Jidong; Guo, Yuan; Zeng, Xiaoqing; Zhou, Shang; Cheng, Jiawei; Zhang, Yuhui

2018-07-05

In this study, a novel aptamer - based fluorescence bio-sensor (aptamer-AuNps) was developed for chiral recognition of arginine (Arg) enantiomers based on aptamer and gold nanoparticles (AuNps). Carboxyfluorescein (FAM) labeled aptamers (Apt) were absorbed on AuNps and their fluorescence intensity could be significantly quenched by AuNps based on fluorescence resonance energy transfer (FRET). Once d-Arg or l-Arg were added into the above solution, the aptamer specifically bind to Arg enantiomers and released from AuNps, so the fluorescence intensity of d-Arg system and l-Arg system were all enhanced. The affinity of Apt to l-Arg is tighter to d-Arg, so the enhanced fluorescence signals of l-Arg system was stronger than d-Arg system. What's more, the enhanced fluorescence were directly proportional to the concentration of d-Arg and l-Arg ranging from 0-300 nM and 0-400 nM with related coefficients of 0.9939 and 0.9952, respectively. Furthermore, the method was successfully applied to detection l-Arg in human urine samples with satisfactory results. Eventually, a simple "OR" logic gate with d-Arg &l-Arg as inputs and AuNps aggregation state as outputs was fabricated, which can help us understand the chiral recognition process deeply. Copyright © 2018 Elsevier B.V. All rights reserved.

Construction of a multiple fluorescence labeling system for use in co-invasion studies of Listeria monocytogenes

DEFF Research Database (Denmark)

Andersen, Jens Bo; Roldgaard, Bent; Lindner, A. B.

2006-01-01

strains at the single cell level by use of fluorescence microscopy. More than 90% of the L. monocytogenes host cells maintained the fluorescence tags for 40 generations. The fluorescence tags did not alter the invasive capacity of the L. monocytogenes cells in a traditional Caco-2 cell invasion assay......, and visual discrimination between invaded bacteria carrying different fluorescent labels inside the cells was possible. Conclusion The constructed fluorescent marker system is stable, easy to use, does not affect the virulence of L. monocytogenes in Caco-2 cell assays, and allows discrimination between...... deviations in the observed capacity for infection when animal models are used. One way to circumvent this problem is to carry out virulence studies as competition assays between 2 or more strains. This, however, requires invasion-neutral markers that enable easy discrimination between the different strains...

[A cell-based detection of ciguatoxin using sodium fluorescence probe].

Science.gov (United States)

Yuan, Jian-hui; Yang, Hui; Tang, Huan-wen; Huang, Wei; Xu, Xin-yun; Liu, Jian-jun; Ke, Yue-bin; Cheng, Jin-quan; Zhuang, Zhi-xiong

2011-04-01

To establish a cell-based detection method of ciguatoxin using fluorescence assay. Mouse neuroblastoma N-2A cells were exposed to ouabain and veratridine and different concentrations of standard ciguatoxin samples (P-CTX-1) to establish the curvilinear relationship between the toxin dosage and fluorescence intensity using the sodium fluorescence probe CoroNaTM Green. The toxicity curvilinear relationship was also generated between the toxin dosage and cell survival using CCK-8 method. Based on these standard curves, the presence of ciguatoxin was detected in 33 samples of deep-sea coral fish. A correlation was found between the detection results of cell-based fluorescence assay and cytotoxicity assay, whose detection limit reached 103 g/ml and 1012 g/ml, respectively. The cell-based fluorescent assay sensitivity showed a higher sensitivity than cytotoxicity assay with a 2-4 h reduction of the detection time. The cell-based fluorescent assay can quickly and sensitively detect ciguatoxin and may serve as a good option for preliminary screening of the toxin.

Fluorescence spectroscopy as a tool for quality assessment of humic substances

Science.gov (United States)

Boguta, Patrycja

2016-04-01

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

A virus-MIPs fluorescent sensor based on FRET for highly sensitive detection of JEV.

Science.gov (United States)

Liang, Caishuang; Wang, Huan; He, Kui; Chen, Chunyan; Chen, Xiaoming; Gong, Hang; Cai, Changqun

2016-11-01

Major stumbling blocks in the recognition and detection of virus are the unstable biological recognition element or the complex detection means. Here a fluorescent sensor based on virus-molecular imprinted polymers (virus-MIPs) was designed for specific recognition and highly sensitive detection of Japanese encephalitis virus (JEV). The virus-MIPs were anchored on the surface of silica microspheres modified by fluorescent dye, pyrene-1-carboxaldehyde (PC). The fluorescence intensity of PC can be enhanced by the principle of fluorescence resonance energy transfer (FRET), where virus acted as energy donor and PC acted as energy acceptor. The enhanced fluorescence intensity was proportional to the concentration of virus in the range of 24-960pM, with a limit of detection (LOD, 3σ) of 9.6pM, and the relative standard deviation was 1.99%. In additional, the specificity study confirmed the resultant MIPs has high-selectivity for JEV. This sensor would become a new key for the detection of virus because of its high sensitive, simple operation, high stability and low cost. Copyright © 2016. Published by Elsevier B.V.

A Dansyl-Rhodamine Based Fluorescent Probe for Detection of Hg2+ and Cu2.

Science.gov (United States)

Yuan, Shizhuang; Su, Wei; Wang, Enju

2017-09-01

A novel fluorescent probe based on dansyl-appended rhodamine B was developed. The probe can selectively recognize and sense Hg2+ and Cu2+ from other common metal ions by showing unique fluorescence and absorption characteristics. In MeCN/HEPES buffer solution, the probe gives a ratiometric fluorescent response to Hg2+, which was ascribed to the fluorescence resonance energy transfer from dansyl moiety to the ring-opened rhodamine B moiety, while the presence of Cu2+ causes fluorescence quenching. Beside the fluorescence change, the presence of Cu2+ and Hg2+ can induce intensive absorption at about 555 nm, which resulted in a color change from colorless to pink.

Phenotypical characteristics of leukocyte membranes in Chernobyl clean-up workers from Latvia: use of the fluorescent probe ABM

International Nuclear Information System (INIS)

Kalnina, I.; Meirovics, I.; Garbuseva, N.; Bruvere, R.; Heisele, O.; Zvagule, T.; Volrate, A.; Feldmane, G.

2001-01-01

A fluorescent probe - aminoderivative of benzanthrone, AMB (developed at the Riga Technical University, Riga, Latvia) - has been previously shown to localise within the phospholipid bilayer of the cell membrane, and shown to affect the structural and functional properties of peripheral blood mononuclear cells (PBMC). The probe ABM was used to characterise the PBMC membranes of 97 Chernobyl clean-up workers from Latvia. The study was conducted in the years 1997-1998. After addition of the probe to PBMC, fluorescence intensity of ABM (F) was measured, the depolarisation value P was calculated, and emission spectra were recorded. Screening of all individuals showed 5 different patterns of fluorescence spectra. Four of the patterns had never been previously observed in healthy individuals or patients with tuberculosis, multiple sclerosis, oncologic patients, etc., examined by us. The spectral patterns of ABM suspensions were associated with ability of leukocytes to produce interferons, with the levels of immunoglobulins A, G, and M, the concentration of lead in peripheral blood, and with several neurologic diseases. The use of ABM allowed to show phenotypic differences in PBMC between Chernobyl clean-up workers and individuals who had never had professional contact with radioactivity. (author)

Surface recognition and fluorescence sensing of histone by dansyl-appended cyclophane-based resorcinarene trimer.

Science.gov (United States)

Hayashida, Osamu; Ogawa, Naoyuki; Uchiyama, Masaki

2007-11-07

A cyclophane-based resorcinarene trimer (3) bearing a dansyl moiety as an environmentally sensitive fluorophore was prepared by stepwise condensation of a tetraaza[6.1.6.1]paracyclophane skeleton with a dansyl moiety and three resorcinarene derivatives having heptacarboxylic acid residues in this sequence. The dansyl-appended cyclophane exhibited the following fluorescence properties regarding solvent polarity dependency and histone surface recognition: With increasing dioxane contents in dioxane/water solvents, the fluorescence intensity originating from the dansyl moiety of 3 increased along with a concomitant blue shift of the fluorescence maximum (lambdaem). The microenvironmentally sensitive fluorescence properties of dansyl fluorophore were maintained, even when the dansyl moiety was covalently attached to a cyclophane. Most interestingly, the cyclophane-based resorcinarene trimer exhibited recognition and fluorescence sensing capabilities toward histone, a small basic protein of eukaryotic chromatins. The fluorescence intensity originating from 3 increased along with a concomitant blue shift of lambdaem upon the addition of histone, reflecting the formation of 3-histone complexes. A relatively large fluorescence polarization (P) value was obtained for the 3-histone complexes (0.15), reflecting highly restricted conformations of 3, and the obtained P value was much larger than that of 3 alone in aqueous medium (0.07). The binding constant (K) of 3 with histone (unit basis) was estimated to be 2.1 x 106 M-1. On the other hand, upon the addition of acetylated histone (Ac-histone) to an aqueous solution containing 3, the extent of change in fluorescence intensity originating from the dansyl group of 3 was almost negligible, indicating that the electrostatic interactions between 3 and Ac-histone were weak. In addition, the fluorescence spectral changes were also small or negligible upon the addition of other proteins such as albumin, ovalbumin, peanut agglutinin

An eco-friendly molecularly imprinted fluorescence composite material based on carbon dots for fluorescent detection of 4-nitrophenol

International Nuclear Information System (INIS)

Hao, Tongfan; Wei, Xiao; Nie, Yijing; Zhou, Zhiping; Xu, Yeqing; Yan, Yongsheng

2016-01-01

We on report an eco-friendly molecularly imprinted material based on carbon dots (C-dots) via a facile and efficient sol–gel polymerization for selective fluorescence detection of 4-nitrophenol (4-NP). The amino-modified C-dots were firstly synthesized by a hydrothermal process using citric acid as the carbon source and poly(ethyleneimine) as the surface modifier, and then after a sol–gel molecular imprinting process, the molecularly imprinted fluorescence material was obtained. The material (MIP-C-dots) showed strong fluorescence from C-dots and high selectivity due to the presence of a molecular imprint. After the detection conditions were optimized, the relative fluorescence intensity (F_0/F) of MIP-C-dots presented a good linearity with 4-NP concentrations in the linear range of 0.2 − 50 μmol L"-"1 with a detection limit (3σ/k) of 0.06 μmol L"-"1. In addition, the correlation coefficient was 0.9978 and the imprinting factor was 2.76. The method was applicable to the determination of trace 4-NP in Yangtze River water samples and good recoveries from 92.6–107.3 % were obtained. The present study provides a general strategy to fabricate materials based on C-dots with good fluorescence property for selective fluorescence detection of organic pollutants. (author)

Comparison of three fluorescence labeling and tracking methods of endothelial progenitor cells in laser-injured retina

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Hui Shi

2018-04-01

Full Text Available AIM: To compare three kinds of fluorescent probes for in vitro labeling and in vivo tracking of endothelial progenitor cells (EPCs in a mouse model of laser-induced retinal injury. METHODS: EPCs were isolated from human umbilical cord blood mononuclear cells and labeled with three different fluorescent probes: 5-(and-6-carboxyfluorescein diacetate succinimidyl ester (CFSE, 1,1′-dilinoleyl-3,3,3′,3′-tetramethylindo-carbocyanine perchlorate linked acetylated low-density lipoprotein (DiI-AcLDL, and green fluorescent protein (GFP. The fluorescent intensity of EPCs was examined by confocal microscopy. Survival rate of labeled EPCs was calculated with trypan blue staining, and their adhesive capability was assessed. A mouse model of retinal injury was induced by laser, and EPCs were injected into the vitreous cavity. Frozen section and fluorescein angiography on flat-mounted retinal samples was employed to track the labeled EPCs in vivo. RESULTS: EPCs labeled with CFSE and DiI-AcLDL exhibited an intense green and red fluorescence at the beginning; the fluorescence intensity decreased gradually to 20.23% and 49.99% respectively, after 28d. On the contrary, the florescent intensity of GFP-labeled EPCs increased in a time-dependent manner. All labeled EPCs showed normal morphology and no significant change in survival and adhesive capability. In the mouse model, transplantation of EPCs showed a protective effect against retinal injury. EPCs labeled with CFSE and DiI-AcLDL were successfully tracked in mice during the development of retinal injury and repair; however, GFP-labeled EPCs were not detected in the laser-injured mouse retina. CONCLUSION: The three fluorescent markers used in this study have their own set of advantages and disadvantages. CFSE and DiI-AcLDL are suitable for short-term EPC-labeling, while GFP should be used for long-term labeling. The choice of fluorescent markers should be guided by the purpose of the study.

Endogenous Fluorescence Signatures in Living Pluripotent Stem Cells Change with Loss of Potency

Science.gov (United States)

Squirrell, Jayne M.; Fong, Jimmy J.; Ariza, Carlos A.; Mael, Amber; Meyer, Kassondra; Shevde, Nirupama K.; Roopra, Avtar; Lyons, Gary E.; Kamp, Timothy J.; Eliceiri, Kevin W.; Ogle, Brenda M.

2012-01-01

The therapeutic potential of stem cells is limited by the non-uniformity of their phenotypic state. Thus it would be advantageous to noninvasively monitor stem cell status. Driven by this challenge, we employed multidimensional multiphoton microscopy to quantify changes in endogenous fluorescence occurring with pluripotent stem cell differentiation. We found that global and cellular-scale fluorescence lifetime of human embryonic stem cells (hESC) and murine embryonic stem cells (mESC) consistently decreased with differentiation. Less consistent were trends in endogenous fluorescence intensity with differentiation, suggesting intensity is more readily impacted by nuances of species and scale of analysis. What emerges is a practical and accessible approach to evaluate, and ultimately enrich, living stem cell populations based on changes in metabolism that could be exploited for both research and clinical applications. PMID:22952742

Fluorescence cell imaging and manipulation using conventional halogen lamp microscopy.

Directory of Open Access Journals (Sweden)

Kazuo Yamagata

Full Text Available Technologies for vitally labeling cells with fluorescent dyes have advanced remarkably. However, to excite fluorescent dyes currently requires powerful illumination, which can cause phototoxic damage to the cells and increases the cost of microscopy. We have developed a filter system to excite fluorescent dyes using a conventional transmission microscope equipped with a halogen lamp. This method allows us to observe previously invisible cell organelles, such as the metaphase spindle of oocytes, without causing phototoxicity. Cells remain healthy even after intensive manipulation under fluorescence observation, such as during bovine, porcine and mouse somatic cell cloning using nuclear transfer. This method does not require expensive epifluorescence equipment and so could help to reduce the science gap between developed and developing countries.

Studying atomic-resolution by X-ray fluorescence holography

International Nuclear Information System (INIS)

Gao Hongyi; Chen Jianwen; Xie Honglan; Zhu Huafeng; Li Ruxin; Xu Zhizhan

2005-01-01

In this work, the results of numerical simulations of X-ray fluorescence holograms and the reconstructed atomic images for Fe single crystal are given. The influences of the recording angles ranges and the polarization effect on the reconstruction of the atomic images are discussed. The process for removing twin images by multiple energy fluorescence holography and expanding the energy range of the incident X-rays to improve the resolution of the reconstructed images is presented

In vivo fluorescence imaging of hepatocellular carcinoma using a novel GPC3-specific aptamer probe

Science.gov (United States)

Zhao, Menglong; Dong, Lili; Liu, Zhuang; Yang, Shuohui

2018-01-01

Background Glypican-3 (GPC3) is highly expressed in most of the hepatocellular carcinomas (HCCs), even in small HCCs. It may be used as a potential biomarker for early detection of HCC. The aptamer is a promising targeting agent with unique advantages over antibody. This study was to introduce a novel GPC3 specific aptamer (AP613-1), to verify its specific binding property in vitro, and to evaluate its targeting efficiency in vivo by performing near-infrared (NIR) fluorescence imaging on an HCC xenograft model. Methods AP613-1 was generated from the systematic evolution of ligands by exponential enrichment. Flow cytometry and aptamer-based immunofluorescence imaging were performed to verify the binding affinity of AP613-1 to GPC3 in vitro. NIR Fluorescence images of nude mice with unilateral (n=12) and bilateral (n=4) subcutaneous xenograft tumors were obtained. Correlation between the tumor fluorescence intensities in vivo and ex vivo was analyzed. Results AP613-1 could specifically bind to GPC3 in vitro. In vivo and ex vivo tumors, fluorescence intensities were in excellent correlation (Pfluorescence intensity is significantly higher in tumors given Alexa Fluor 750 (AF750) labeled AP613-1 than in those given AF750 labeled initial ssDNA library both in vivo (Pfluorescence intensities than A549 tumors both in vivo (P=0.016) and ex vivo (P=0.004). Conclusions AP613-1 displays a specific binding affinity to GPC3 positive HCC. Fluorescently labeled AP613-1 could be used as an imaging probe to subcutaneous HCC in xenograft models. PMID:29675356

Using Continuous In-situ Measurement of Fluorescence to Reveal Hot Spots and Hot Moments of Dissolved Organic Matter Dynamics in a Forested Watershed

Science.gov (United States)

Ryan, K. A.; Hosen, J. D.; Raymond, P. A.; Stubbins, A.; Shanley, J. B.

2017-12-01

River systems serve as net carbon exporters from land to the ocean, fueling downstream aquatic ecosystem food webs. Fluorescence signatures of aquatic organic matter can be used as a proxy for dissolved organic carbon (DOC) concentration and can characterize DOC composition, reactivity, and source to improve our understanding of ecological processes. In-situ measurement of fluorescence using fifteen-minute interval data logging allows greater temporal resolution than laboratory studies. However, in-situ data must be corrected for interferences from temperature, absorbance and turbidity changes occurring in the field. We installed multiparameter water quality sondes (Eureka Mantas) and in-situ fluorometers (Turner Designs Cyclops) at sites nested within streams and riparian zones in the Sleepers River Research Watershed in Vermont in 2017. We coupled these measurements with simultaneous intensive field sampling campaigns and laboratory analysis of DOC and fluorescence Excitation-Emission Matrices. The data loggers from the nested sites recorded fluorescence peaks responding to discharge events and tracked changes in fluorescence occurring from upstream to downstream sites. Laboratory results confirm a nonlinear, hysteretic relationship between discharge and DOC where peak DOC lags peak discharge. This hysteresis is predicted to be controlled by multiple flow paths and DOC sources (i.e. groundwater, overland flow). We conclude that continuous in-situ records of river water fluorescence can be used to inform ecological processes and test new hypotheses concerning dissolved organic matter dynamics in watersheds.

Optimizing plasmon-enhanced fluorescence with nonlocal metallic nanospheres

DEFF Research Database (Denmark)

Tserkezis, Christos; Stefanou, Nikolaos; Wubs, Martijn

, through the recent Generalized Nonlocal Optical Response (GNOR) theory, the concurrent contribution of modal shifts and nonradiative losses, together with a reduced emitter excitation rate due to the decreased field intensity, lead always to a strong reduction of fluorescence (see Fig. 1). Finally, we...... identify situations where the common, intuitive recipe of tuning the NP modes to match λem can in fact lead to strong fluorescence quenching, instead of the anticipated enhancement. Our results highlight the necessity for careful modeling and design of plasmon-field-enhancement based applications....

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

Energy Technology Data Exchange (ETDEWEB)

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

1999-08-01

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

Fluorescence fluctuation of Rhodamine 6G dye for high repetition rate laser excitation

International Nuclear Information System (INIS)

Singh, Nageshwar; Patel, Hemant K.; Dixit, S.K.; Vora, H.S.

2013-01-01

In this paper, fluorescence from Rhodamine 6G dye for stationary and flowing liquid medium, excited by copper vapor laser, operating at 6 kHz pulse repetition frequency, was investigated. Large fluctuations in spectral width (about 5 nm) and spectral intensity in the fluorescence from stationary dye solution were observed, while fluctuations in the spectral width diminish in a flowing dye medium. However, this increases spectral intensity and slightly red shifts the fluorescence peak emission wavelength. Theoretical analysis was carried out to explain the observed results by incorporating the temperature induced refractive index, beam deflection and spectral variation in stationary dye solution. Numerical analysis of thermal load and contour of temperature in the optical pumped region inside the dye cell in stationary, 0.2 and 1.5 m/s flow velocity was also investigated to support our analysis. - Highlights: ► High repetition rate excitation generates inhomogeneity in the gain medium. ► Fluorescence of Rhodamine 6G in stationary and flowing medium was carried out. ► Fluorescence fluctuations lessen in flowing medium in contrast to stationary medium. ► Our theoretical and numerical analysis enlightens the experimented outcome trend.

vuv fluorescence from selective high-order multiphoton excitation of N2

International Nuclear Information System (INIS)

Coffee, Ryan N.; Gibson, George N.

2004-01-01

Recent fluorescence studies suggest that ultrashort pulse laser excitation may be highly selective. Selective high-intensity laser excitation holds important consequences for the physics of multiphoton processes. To establish the extent of this selectivity, we performed a detailed comparative study of the vacuum ultraviolet fluorescence resulting from the interaction of N 2 and Ar with high-intensity infrared ultrashort laser pulses. Both N 2 and Ar reveal two classes of transitions, inner-valence ns ' l ' . From their pressure dependence, we associate each transition with either plasma or direct laser excitation. Furthermore, we qualitatively confirm such associations with the time dependence of the fluorescence signal. Remarkably, only N 2 presents evidence of direct laser excitation. This direct excitation produces ionic nitrogen fragments with inner-valence (2s) holes, two unidentified transitions, and one molecular transition, the N 2 + :X 2 Σ g + 2 Σ u + . We discuss these results in the light of a recently proposed model for multiphoton excitation

Creating infinite contrast in fluorescence microscopy by using lanthanide centered emission

DEFF Research Database (Denmark)

R. Carro-Temboury, Miguel; Arppe, Riikka Matleena; Hempel, Casper

2017-01-01

The popularity of fluorescence microscopy arises from the inherent mode of action, where the fluorescence emission from probes is used to visualize selected features on a presumed dark background. However, the background is rarely truly dark, and image processing and analysis is needed to enhance...... the fluorescent signal that is ascribed to the selected feature. The image acquisition is facilitated by using considerable illumination, bright probes at a relatively high concentration in order to make the fluorescent signal significantly more intense than the background signal. Here, we present two methods......, while method II resolves the fluorescent signal by subtracting a background calculated via the gradient. Both methods improve signal-to-background ratio significantly and we suggest that spectral imaging of lanthanide-centered emission can be used as a tool to obtain absolute contrast in bioimaging....

Fluorescent single walled nanotube/silica composite materials

Science.gov (United States)

Dattelbaum, Andrew M.; Gupta, Gautam; Duque, Juan G.; Doorn, Stephen K.; Hamilton, Christopher E.; DeFriend Obrey, Kimberly A.

2013-03-12

Fluorescent composites of surfactant-wrapped single-walled carbon nanotubes (SWNTs) were prepared by exposing suspensions of surfactant-wrapped carbon nanotubes to tetramethylorthosilicate (TMOS) vapor. Sodium deoxycholate (DOC) and sodium dodecylsulphate (SDS) were the surfactants. No loss in emission intensity was observed when the suspension of DOC-wrapped SWNTs were exposed to the TMOS vapors, but about a 50% decrease in the emission signal was observed from the SDS-wrapped SWNTs nanotubes. The decrease in emission was minimal by buffering the SDS/SWNT suspension prior to forming the composite. Fluorescent xerogels were prepared by adding glycerol to the SWNT suspensions prior to TMOS vapor exposure, followed by drying the gels. Fluorescent aerogels were prepared by replacing water in the gels with methanol and then exposing them to supercritical fluid drying conditions. The aerogels can be used for gas sensing.

The application of anti-ESAT-6 monoclonal antibody fluorescent probe in ex vivo near-infrared fluorescence imaging in mice with pulmonary tuberculosis.

Science.gov (United States)

Feng, Feng; Zhang, Haoling; Zhu, Zhaoqin; Li, Cong; Shi, Yuxin; Zhang, Zhiyong

2014-09-01

Here, we aimed to assess the feasibility of anti-ESAT-6 monoclonal antibody (mAb) coupling with IR783 and rhodamine fluorescent probe in the detection of ESAT-6 expression in tuberculosis tissue of mice using near-infrared fluorescence imaging. IR783 and rhodamine were conjugated to the anti-ESAT-6 mAb or IgG. Mice in the experimental group were injected with fluorescence-labeled mAb probe, and mice in the control group were injected with fluorescence-labeled non-specific IgG antibody. Twenty-four hours later, the lung tissue of mice was examined using ex vivo near-infrared fluorescence imaging. In addition, the contrast-to-noise ratio (CNR) was calculated by measuring the signal intensities of the pulmonary lesions, normal lung tissue and background noise. The frozen lung tissue section was examined under fluorescence microscopy and compared with hemoxylin and eosin (HE) staining. The ex vivo near-infrared fluorescence imaging showed that the fluorescence signal in the lung tuberculosis lesions in the experimental group was significantly enhanced, whereas there was only a weak fluorescence signal or even no fluorescence signal in the control group. CNR values were 64.40 ± 7.02 (n = 6) and 8.75 ± 3.87 (n = 6), respectively (t = 17.01, p fluorescence accumulation distribution detected under fluorescence microscopy was consistent with HE staining of the tuberculosis region. In conclusion, anti-ESAT-6 mAb fluorescent probe could target and be applied in specific ex vivo imaging of mice tuberculosis, and may be of further use in tuberculosis in living mice. Copyright © 2013 John Wiley & Sons, Ltd.

Absorption and fluorescence studies of curcumin bound to liposomes and lymphocytes: effect of γ- irradiation

International Nuclear Information System (INIS)

Kunwar, Amit; Barik, A.; Indira Priyadarsini, K.; Pandey, R.

2006-01-01

Absorption and fluorescence spectral changes in curcumin were employed to follow its binding to liposomes and lymphocytes. The association constants indicated high affinity of curcumin to liposomes. Tumor lymphocytes show mere intense fluorescence of curcumin over the normal lymphocytes. The loss of curcumin in cells after γ-irradiation could be followed by reduction in curcumin fluorescence. The studies indicate that such fluorescence changes can be used as markers to understand the preferential loading of curcumin to cells. (author)

Absorption and fluorescence studies of curcumin bound to liposomes and lymphocytes: effect of {gamma}- irradiation

Energy Technology Data Exchange (ETDEWEB)

Kunwar, Amit; Barik, A; Indira Priyadarsini, K [Radiation and Photochemistry Division, Bhabha Atomic Research Centre, Mumbai (India); Pandey, R [Radiation Biology and Health Sciences Division, Bhabha Atomic Research Centre, Mumbai (India)

2006-01-15

Absorption and fluorescence spectral changes in curcumin were employed to follow its binding to liposomes and lymphocytes. The association constants indicated high affinity of curcumin to liposomes. Tumor lymphocytes show mere intense fluorescence of curcumin over the normal lymphocytes. The loss of curcumin in cells after {gamma}-irradiation could be followed by reduction in curcumin fluorescence. The studies indicate that such fluorescence changes can be used as markers to understand the preferential loading of curcumin to cells. (author)

Compact point-detection fluorescence spectroscopy system for quantifying intrinsic fluorescence redox ratio in brain cancer diagnostics

Science.gov (United States)

Liu, Quan; Grant, Gerald; Li, Jianjun; Zhang, Yan; Hu, Fangyao; Li, Shuqin; Wilson, Christy; Chen, Kui; Bigner, Darell; Vo-Dinh, Tuan

2011-03-01

We report the development of a compact point-detection fluorescence spectroscopy system and two data analysis methods to quantify the intrinsic fluorescence redox ratio and diagnose brain cancer in an orthotopic brain tumor rat model. Our system employs one compact cw diode laser (407 nm) to excite two primary endogenous fluorophores, reduced nicotinamide adenine dinucleotide, and flavin adenine dinucleotide. The spectra were first analyzed using a spectral filtering modulation method developed previously to derive the intrinsic fluorescence redox ratio, which has the advantages of insensitivty to optical coupling and rapid data acquisition and analysis. This method represents a convenient and rapid alternative for achieving intrinsic fluorescence-based redox measurements as compared to those complicated model-based methods. It is worth noting that the method can also extract total hemoglobin concentration at the same time but only if the emission path length of fluorescence light, which depends on the illumination and collection geometry of the optical probe, is long enough so that the effect of absorption on fluorescence intensity due to hemoglobin is significant. Then a multivariate method was used to statistically classify normal tissues and tumors. Although the first method offers quantitative tissue metabolism information, the second method provides high overall classification accuracy. The two methods provide complementary capabilities for understanding cancer development and noninvasively diagnosing brain cancer. The results of our study suggest that this portable system can be potentially used to demarcate the elusive boundary between a brain tumor and the surrounding normal tissue during surgical resection.

Quantitative analysis by computer controlled X-ray fluorescence spectrometer

International Nuclear Information System (INIS)

Balasubramanian, T.V.; Angelo, P.C.

1981-01-01

X-ray fluorescence spectroscopy has become a widely accepted method in the metallurgical field for analysis of both minor and major elements. As encountered in many other analytical techniques, the problem of matrix effect generally known as the interelemental effects is to be dealt with effectively in order to make the analysis accurate. There are several methods by which the effects of matrix on the analyte are minimised or corrected for and the mathematical correction is one among them. In this method the characteristic secondary X-ray intensities are measured from standard samples and correction coefficients. If any, for interelemental effects are evaluated by mathematical calculations. This paper describes attempts to evaluate the correction coefficients for interelemental effects by multiple linear regression programmes using a computer for the quantitative analysis of stainless steel and a nickel base cast alloy. The quantitative results obtained using this method for a standard stainless steel sample are compared with the given certified values. (author)

Multispectral fluorescence imaging techniques for nondestructive food safety inspection

Science.gov (United States)

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

2004-03-01

The use of spectral sensing has gained acceptance as a rapid means for nondestructive inspection of postharvest food produce. Current technologies generally use color or a single wavelength camera technology. The applicability and sensitivity of these techniques can be expanded through the use of multiple wavelengths. Reflectance in the Vis/NIR is the prevalent spectral technique. Fluorescence, compared to reflectance, is regarded as a more sensitive technique due to its dynamic responses to subtle changes in biological entities. Our laboratory has been exploring fluorescence as a potential means for detection of quality and wholesomeness of food products. Applications of fluorescence sensing require an understanding of the spectral characteristics emanating from constituents and potential contaminants. A number of factors affecting fluorescence emission characteristics are discussed. Because of relatively low fluorescence quantum yield from biological samples, a system with a powerful pulse light source such as a laser coupled with a gated detection device is used to harvest fluorescence, in the presence of ambient light. Several fluorescence sensor platforms developed in our laboratory, including hyperspectral imaging, and laser-induced fluorescence (LIF) and steady-state fluorescence imaging systems with multispectral capabilities are presented. We demonstrate the potential uses of recently developed fluorescence imaging platforms in food safety inspection of apples contaminated with animal feces.

Fluorescence spectroscopy for neoplasms control

Science.gov (United States)

Bratchenko, I. A.; Kristoforova, Yu. A.; Myakinin, O. O.; Artemyev, D. N.; Kozlov, S. V.; Moryatov, A. A.; Zakharov, V. P.

2016-04-01

Investigation of malignant skin tumors diagnosis was performed involving two setups for native tissues fluorescence control in visible and near infrared regions. Combined fluorescence analysis for skin malignant melanomas and basal cell carcinomas was performed. Autofluorescence spectra of normal skin and oncological pathologies stimulated by 457 nm and 785 nm lasers were registered for 74 skin tissue samples. Spectra of 10 melanomas and 27 basal cell carcinomas were registered ex vivo. Skin tumors analysis was made on the basis of autofluorescence spectra intensity and curvature for analysis of porphyrins, lipo-pigments, flavins and melanin. Separation of melanomas and basal cell carcinomas was performed on the basis of discriminant analysis. Overall accuracy of basal cell carcinomas and malignant melanomas separation in current study reached 86.5% with 70% sensitivity and 92.6% specificity.

Time-Resolved Fluorescence Immunoassay for C-Reactive Protein Using Colloidal Semiconducting Nanoparticles

Directory of Open Access Journals (Sweden)

Pekka Hänninen

2011-11-01

Full Text Available Besides the typical short-lived fluorescence with decay times in the nanosecond range, colloidal II/VI semiconductor nanoparticles dispersed in buffer also possess a long-lived fluorescence component with decay times in the microsecond range. Here, the signal intensity of the long-lived luminescence at microsecond range is shown to increase 1,000-fold for CdTe nanoparticles in PBS buffer. This long-lived fluorescence can be conveniently employed for time-gated fluorescence detection, which allows for improved signal-to-noise ratio and thus the use of low concentrations of nanoparticles. The detection principle is demonstrated with a time-resolved fluorescence immunoassay for the detection of C-reactive protein (CRP using CdSe-ZnS nanoparticles and green light excitation.

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

Science.gov (United States)

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

2013-09-01

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

Recent Progress on Plasmon-Enhanced Fluorescence

Directory of Open Access Journals (Sweden)

Dong Jun

2015-12-01

Full Text Available The optically generated collective electron density waves on metal–dielectric boundaries known as surface plasmons have been of great scientific interest since their discovery. Being electromagnetic waves on gold or silver nanoparticle’s surface, localised surface plasmons (LSP can strongly enhance the electromagnetic field. These strong electromagnetic fields near the metal surfaces have been used in various applications like surface enhanced spectroscopy (SES, plasmonic lithography, plasmonic trapping of particles, and plasmonic catalysis. Resonant coupling of LSPs to fluorophore can strongly enhance the emission intensity, the angular distribution, and the polarisation of the emitted radiation and even the speed of radiative decay, which is so-called plasmon enhanced fluorescence (PEF. As a result, more and more reports on surface-enhanced fluorescence have appeared, such as SPASER-s, plasmon assisted lasing, single molecule fluorescence measurements, surface plasmoncoupled emission (SPCE in biological sensing, optical orbit designs etc. In this review, we focus on recent advanced reports on plasmon-enhanced fluorescence (PEF. First, the mechanism of PEF and early results of enhanced fluorescence observed by metal nanostructure will be introduced. Then, the enhanced substrates, including periodical and nonperiodical nanostructure, will be discussed and the most important factor of the spacer between molecule and surface and wavelength dependence on PEF is demonstrated. Finally, the recent progress of tipenhanced fluorescence and PEF from the rare-earth doped up-conversion (UC and down-conversion (DC nanoparticles (NPs are also commented upon. This review provides an introduction to fundamentals of PEF, illustrates the current progress in the design of metallic nanostructures for efficient fluorescence signal amplification that utilises propagating and localised surface plasmons.

In vivo multiphoton tomography and fluorescence lifetime imaging of human brain tumor tissue.

Science.gov (United States)

Kantelhardt, Sven R; Kalasauskas, Darius; König, Karsten; Kim, Ella; Weinigel, Martin; Uchugonova, Aisada; Giese, Alf

2016-05-01

High resolution multiphoton tomography and fluorescence lifetime imaging differentiates glioma from adjacent brain in native tissue samples ex vivo. Presently, multiphoton tomography is applied in clinical dermatology and experimentally. We here present the first application of multiphoton and fluorescence lifetime imaging for in vivo imaging on humans during a neurosurgical procedure. We used a MPTflex™ Multiphoton Laser Tomograph (JenLab, Germany). We examined cultured glioma cells in an orthotopic mouse tumor model and native human tissue samples. Finally the multiphoton tomograph was applied to provide optical biopsies during resection of a clinical case of glioblastoma. All tissues imaged by multiphoton tomography were sampled and processed for conventional histopathology. The multiphoton tomograph allowed fluorescence intensity- and fluorescence lifetime imaging with submicron spatial resolution and 200 picosecond temporal resolution. Morphological fluorescence intensity imaging and fluorescence lifetime imaging of tumor-bearing mouse brains and native human tissue samples clearly differentiated tumor and adjacent brain tissue. Intraoperative imaging was found to be technically feasible. Intraoperative image quality was comparable to ex vivo examinations. To our knowledge we here present the first intraoperative application of high resolution multiphoton tomography and fluorescence lifetime imaging of human brain tumors in situ. It allowed in vivo identification and determination of cell density of tumor tissue on a cellular and subcellular level within seconds. The technology shows the potential of rapid intraoperative identification of native glioma tissue without need for tissue processing or staining.

Fluorescence imaging of tryptophan and collagen cross-links to evaluate wound closure ex vivo

Science.gov (United States)

Wang, Ying; Ortega-Martinez, Antonio; Farinelli, Bill; Anderson, R. R.; Franco, Walfre

2016-02-01

Wound size is a key parameter in monitoring healing. Current methods to measure wound size are often subjective, time-consuming and marginally invasive. Recently, we developed a non-invasive, non-contact, fast and simple but robust fluorescence imaging (u-FEI) method to monitor the healing of skin wounds. This method exploits the fluorescence of native molecules to tissue as functional and structural markers. The objective of the present study is to demonstrate the feasibility of using variations in the fluorescence intensity of tryptophan and cross-links of collagen to evaluate proliferation of keratinocyte cells and quantitate size of wound during healing, respectively. Circular dermal wounds were created in ex vivo human skin and cultured in different media. Two serial fluorescence images of tryptophan and collagen cross-links were acquired every two days. Histology and immunohistology were used to validate correlation between fluorescence and epithelialization. Images of collagen cross-links show fluorescence of the exposed dermis and, hence, are a measure of wound area. Images of tryptophan show higher fluorescence intensity of proliferating keratinocytes forming new epithelium, as compared to surrounding keratinocytes not involved in epithelialization. These images are complementary since collagen cross-links report on structure while tryptophan reports on function. HE and immunohistology show that tryptophan fluorescence correlates with newly formed epidermis. We have established a fluorescence imaging method for studying epithelialization processes during wound healing in a skin organ culture model, our approach has the potential to provide a non-invasive, non-contact, quick, objective and direct method for quantitative measurements in wound healing in vivo.

Fluorescence of Alexa fluor dye tracks protein folding.

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Simon Lindhoud

Full Text Available Fluorescence spectroscopy is an important tool for the characterization of protein folding. Often, a protein is labeled with appropriate fluorescent donor and acceptor probes and folding-induced changes in Förster Resonance Energy Transfer (FRET are monitored. However, conformational changes of the protein potentially affect fluorescence properties of both probes, thereby profoundly complicating interpretation of FRET data. In this study, we assess the effects protein folding has on fluorescence properties of Alexa Fluor 488 (A488, which is commonly used as FRET donor. Here, A488 is covalently attached to Cys69 of apoflavodoxin from Azotobacter vinelandii. Although coupling of A488 slightly destabilizes apoflavodoxin, the three-state folding of this protein, which involves a molten globule intermediate, is unaffected. Upon folding of apoflavodoxin, fluorescence emission intensity of A488 changes significantly. To illuminate the molecular sources of this alteration, we applied steady state and time-resolved fluorescence techniques. The results obtained show that tryptophans cause folding-induced changes in quenching of Alexa dye. Compared to unfolded protein, static quenching of A488 is increased in the molten globule. Upon populating the native state both static and dynamic quenching of A488 decrease considerably. We show that fluorescence quenching of Alexa Fluor dyes is a sensitive reporter of conformational changes during protein folding.

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

Science.gov (United States)

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

1994-01-01

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

Fluorescence dye tagging scheme for mercury quantification and speciation

Science.gov (United States)

Jiao, Hong; Catterall, Hannah

2015-09-22

A fluorescent dye or fluorophore capable of forming complexes with mercury comprises 6,8-difluoro-7-hydroxy-2-oxo-2H-chromene-3-carboxylate amide, wherein the amide is formed by reacting the succinimidyl ester (Pacific Blue.TM.) with an amino acid containing a thiol group, such as cysteine or glutathione. Mercury complexes of the fluorophore fluoresce when excited by a UV or violet laser diode, and the detected intensity can be calibrated to quantify the concentration of mercury in a sample reacted with the fluorophore.

Simultaneous neuron- and astrocyte-specific fluorescent marking

Energy Technology Data Exchange (ETDEWEB)

Schulze, Wiebke [Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Hayata-Takano, Atsuko [Molecular Research Center for Children' s Mental Development, United Graduate School of Child Development, Osaka University, Kanazawa University, Hamamatsu University School of Medicine, Chiba University and University of Fukui, 2-2 Yamadaoka, Suita, Osaka 565-0871 (Japan); Kamo, Toshihiko [Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Nakazawa, Takanobu, E-mail: takanobunakazawa-tky@umin.ac.jp [iPS Cell-based Research Project on Brain Neuropharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Nagayasu, Kazuki [iPS Cell-based Research Project on Brain Neuropharmacology and Toxicology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Kasai, Atsushi; Seiriki, Kaoru [Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Interdisciplinary Program for Biomedical Sciences, Institute for Academic Initiatives, Osaka University, 1-1 Yamadaoka, Suita, Osaka 565-0871 (Japan); Shintani, Norihito [Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Ago, Yukio [Laboratory of Medicinal Pharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); Farfan, Camille [Laboratory of Molecular Neuropharmacology, Graduate School of Pharmaceutical Sciences, Osaka University, 1-6 Yamadaoka, Suita, Osaka 565-0871 (Japan); and others

2015-03-27

Systematic and simultaneous analysis of multiple cell types in the brain is becoming important, but such tools have not yet been adequately developed. Here, we aimed to generate a method for the specific fluorescent labeling of neurons and astrocytes, two major cell types in the brain, and we have developed lentiviral vectors to express the red fluorescent protein tdTomato in neurons and the enhanced green fluorescent protein (EGFP) in astrocytes. Importantly, both fluorescent proteins are fused to histone 2B protein (H2B) to confer nuclear localization to distinguish between single cells. We also constructed several expression constructs, including a tandem alignment of the neuron- and astrocyte-expression cassettes for simultaneous labeling. Introducing these vectors and constructs in vitro and in vivo resulted in cell type-specific and nuclear-localized fluorescence signals enabling easy detection and distinguishability of neurons and astrocytes. This tool is expected to be utilized for the simultaneous analysis of changes in neurons and astrocytes in healthy and diseased brains. - Highlights: • We develop a method for the specific fluorescent labeling of neurons and astrocytes. • Neuron-specific labeling is achieved using Scg10 and synapsin promoters. • Astrocyte-specific labeling is generated using the minimal GFAP promoter. • Nuclear localization of fluorescent proteins is achieved with histone 2B protein.

Simultaneous neuron- and astrocyte-specific fluorescent marking

International Nuclear Information System (INIS)

Schulze, Wiebke; Hayata-Takano, Atsuko; Kamo, Toshihiko; Nakazawa, Takanobu; Nagayasu, Kazuki; Kasai, Atsushi; Seiriki, Kaoru; Shintani, Norihito; Ago, Yukio; Farfan, Camille

2015-01-01

Systematic and simultaneous analysis of multiple cell types in the brain is becoming important, but such tools have not yet been adequately developed. Here, we aimed to generate a method for the specific fluorescent labeling of neurons and astrocytes, two major cell types in the brain, and we have developed lentiviral vectors to express the red fluorescent protein tdTomato in neurons and the enhanced green fluorescent protein (EGFP) in astrocytes. Importantly, both fluorescent proteins are fused to histone 2B protein (H2B) to confer nuclear localization to distinguish between single cells. We also constructed several expression constructs, including a tandem alignment of the neuron- and astrocyte-expression cassettes for simultaneous labeling. Introducing these vectors and constructs in vitro and in vivo resulted in cell type-specific and nuclear-localized fluorescence signals enabling easy detection and distinguishability of neurons and astrocytes. This tool is expected to be utilized for the simultaneous analysis of changes in neurons and astrocytes in healthy and diseased brains. - Highlights: • We develop a method for the specific fluorescent labeling of neurons and astrocytes. • Neuron-specific labeling is achieved using Scg10 and synapsin promoters. • Astrocyte-specific labeling is generated using the minimal GFAP promoter. • Nuclear localization of fluorescent proteins is achieved with histone 2B protein

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

Science.gov (United States)

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

2016-05-01

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

Differentiating between heat pain intensities: the combined effect of multiple autonomic parameters.

Science.gov (United States)

Treister, Roi; Kliger, Mark; Zuckerman, Galit; Goor Aryeh, Itay; Eisenberg, Elon

2012-09-01

Although it is well known that pain induces changes in autonomic parameters, the extent to which these changes correlate with the experience of pain is under debate. The aim of the present study was to compare a combination of multiple autonomic parameters and each parameter alone in their ability to differentiate among 4 categories of pain intensity. Tonic heat stimuli (1minute) were individually adjusted to induce no pain, low, medium, and high pain in 45 healthy volunteers. Electrocardiogram, photoplethysmogram, and galvanic skin response were recorded, and the following parameters were calculated: heart rate; heart rate variability-high frequency (0.15 to 0.4Hz) spectral power; skin conductance level; number of skin conduction fluctuations; and photoplethysmographic pulse wave amplitude. A combination of parameters was created by fitting an ordinal cumulative logit model to the data and using linear coefficients of the model. Friedman test with post-hoc Wilcoxon test were used to compare between pain intensity categories for every parameter alone and for their linear combination. All of the parameters successfully differentiated between no pain and all other pain categories. However, none of the parameters differentiated between all 3 pain categories (i.e., low and medium; medium and high; low and high). In contrast, the linear combination of parameters significantly differentiated not only between pain and no pain, but also between all pain categories (Ppain assessment. Copyright © 2012 International Association for the Study of Pain. Published by Elsevier B.V. All rights reserved.

Fluorescence and chemiluminescence behavior of distyrylbenzene bearing two arms of dipicolylaminomethyl groups: Interactions with zinc ion and ATP

Science.gov (United States)

Motoyoshiya, Jiro; Wada, Jun-ya; Itoh, Keiko; Wakabayashi, Kazuaki; Maruyama, Takayuki; Ono, Kazuki; Fukasawa, Kota; Fujimoto, Tetsuya; Akaiwa, Yuji; Nonaka, Eiji

2018-04-01

The absorption and fluorescence spectral study of the distyrylbenzene bearing two arms of the dipicolylaminomethyl groups, the effective ligands for Zn2+, was studied in the presence of Zn2+ and ATP. Upon complexation of the distyrylbenzene with zinc ions in acetonitrile, enhancement of the fluorescence intensity was observed due to inhibition of intramolecular PET (photo-induced electron transfer) quenching, but no effect was found in aqueous media because the equilibrium laid to the free form of the ligands. In contrast, the addition of ATP disodium salt was effective to enhance the fluorescence intensity of the combination of the distyrylbenzne and Zn2+ in aqueous media. This assembly was applied to the peroxyoxalate chemiluminescence system and a significant increase in the intensity was observed, which provides a potential detection for ATP by chemiluminescence.

Laser induced uranium fluorescence as an analytical method

International Nuclear Information System (INIS)

Krutman, I.

1985-01-01

A laser induced fluorescence system was developed to measure uranium trace level amounts in aqueous solution with reliable and simple materials and electronics. A nitrogen pulsed laser was built with the storage energy capacitor directly coupled to laser tube electrodes as a transmission line device. This laser operated at 3Hz repetition rate with peak intensity around 21 Kw and temporal width of 4.5 x 10 -9 s. A sample compartment made of rigid PVC and a photomultiplier housing of aluminium were constructed and assembled forming a single integrated device. As a result of this prototype system we made several analytical measurements with U dissolved in nitric acid to obtain a calibration curve. We obtained a straight line from a plot of U concentration versus fluorescence intensity fitted by a least square method that produced a regression coefficient of 0.994. The lower limit of U determination was 30 ppb -+ 3.5%. (Author) [pt

Nuclear resonance fluorescence of {sup 203,205}Tl

Energy Technology Data Exchange (ETDEWEB)

Pfeifer, Fabian; Fritzsche, Matthias; Pietralla, Norbert; Savran, Deniz; Weller, Henry; Zweidinger, Markus [Institut fuer Kernphysik, Technische Universitaet, Darmstadt (Germany); Rusev, Gencho; Tonchev, Anton P.; Tornow, Werner [Triangle Universities Nuclear Laboratory, Duke University, Durham (United States); Zilges, Andreas [Institut fuer Kernphysik, Universitaet Koeln (Germany)

2009-07-01

In order to investigate the dipole strength distribution in Thalium isotopes we have studied Nuclear Resonance Fluorescence of a sample composed of natural Thallium (consisting of 30% {sup 203}Tl and 70% {sup 205}Tl). Unpolarized bremsstrahlung with photo energies up to 7.5 MeV was used at the High Intensity Photon Setup (HIPS) at S-DALINAC at the IKP Darmstadt. 24 fluorescent {gamma}-ray transitions were observed, 19 of them for the first time. For the assignment of the polarity of two prominent {gamma}-ray transitions, one at 4.7 MeV and one at 4.9 MeV, the polarized photon beam of the High Intensity {gamma}-ray Source (HI{gamma}S) at Duke University was used. The experiment at HI{gamma}S revealed the existence of a photo-excited state of {sup 205}Tl at an excitation energy of 4.971 MeV that exhibits a transition to the first excited state at 203 keV.

Laser excited fluorescence spectrum of Ho3+:SrF2 single crystal

International Nuclear Information System (INIS)

Lal, Bansi; Ramachandra Rao, D.

1980-01-01

The fluorescence spectrum of Ho 3+ : SrF 2 single crystal excited by the various lines of an Ar + laser, is reported. The three fluorescence groups recorded in the region 5300-7700 A, correspond to the transitions from ( 5 F 4 , 5 S 2 ) to 5 I 8 , 5 F 5 to 5 I 8 , 5 F 3 to 5 I 7 and ( 5 F 4 , 5 S 2 ) to 5 I 7 . Marked changes in the total integrated intensity of the various fluorescence groups with the change in the exciting wavelength are observed. (author)

5-ALA induced fluorescent image analysis of actinic keratosis

Science.gov (United States)

Cho, Yong-Jin; Bae, Youngwoo; Choi, Eung-Ho; Jung, Byungjo

2010-02-01

In this study, we quantitatively analyzed 5-ALA induced fluorescent images of actinic keratosis using digital fluorescent color and hyperspectral imaging modalities. UV-A was utilized to induce fluorescent images and actinic keratosis (AK) lesions were demarcated from surrounding the normal region with different methods. Eight subjects with AK lesion were participated in this study. In the hyperspectral imaging modality, spectral analysis method was utilized for hyperspectral cube image and AK lesions were demarcated from the normal region. Before image acquisition, we designated biopsy position for histopathology of AK lesion and surrounding normal region. Erythema index (E.I.) values on both regions were calculated from the spectral cube data. Image analysis of subjects resulted in two different groups: the first group with the higher fluorescence signal and E.I. on AK lesion than the normal region; the second group with lower fluorescence signal and without big difference in E.I. between two regions. In fluorescent color image analysis of facial AK, E.I. images were calculated on both normal and AK lesions and compared with the results of hyperspectral imaging modality. The results might indicate that the different intensity of fluorescence and E.I. among the subjects with AK might be interpreted as different phases of morphological and metabolic changes of AK lesions.

Theoretical Influence Coefficients For X-Ray Fluorescence Analysis Of Alloys

International Nuclear Information System (INIS)

Okunade, I.O.

2004-01-01

The problem of quantifications in X-ray fluorescence analysis has over the years been narrowed down to matrix effects arising from the presence of other elements in the sample, which may either lead to the reduction or enhancement in the measured intensities of the analytic element. This paper describes a mathematical matrix correction method, which yield certain fundamental coefficients that account for the inter-element effects. The application of these influence coefficients in quantitative analysis however relies on the knowledge of pure element intensities of the analyse element, its mass absorption coefficients (for exciting and fluorescent radiation) of other elements in the sample that are responsible for the matrix effects. The quantification method using these coefficients are thereafter established for binary systems and further extended to multi-component systems such as ternary and quaternary alloys

Plant Cell Imaging Based on Nanodiamonds with Excitation-Dependent Fluorescence

Science.gov (United States)

Su, Li-Xia; Lou, Qing; Jiao, Zhen; Shan, Chong-Xin

2016-09-01

Despite extensive work on fluorescence behavior stemming from color centers of diamond, reports on the excitation-dependent fluorescence of nanodiamonds (NDs) with a large-scale redshift from 400 to 620 nm under different excitation wavelengths are so far much fewer, especially in biological applications. The fluorescence can be attributed to the combined effects of the fraction of sp2-hybridized carbon atoms among the surface of the fine diamond nanoparticles and the defect energy trapping states on the surface of the diamond. The excitation-dependent fluorescent NDs have been applied in plant cell imaging for the first time. The results reported in this paper may provide a promising route to multiple-color bioimaging using NDs.

Plant Cell Imaging Based on Nanodiamonds with Excitation-Dependent Fluorescence.

Science.gov (United States)

Su, Li-Xia; Lou, Qing; Jiao, Zhen; Shan, Chong-Xin

2016-12-01

Despite extensive work on fluorescence behavior stemming from color centers of diamond, reports on the excitation-dependent fluorescence of nanodiamonds (NDs) with a large-scale redshift from 400 to 620 nm under different excitation wavelengths are so far much fewer, especially in biological applications. The fluorescence can be attributed to the combined effects of the fraction of sp(2)-hybridized carbon atoms among the surface of the fine diamond nanoparticles and the defect energy trapping states on the surface of the diamond. The excitation-dependent fluorescent NDs have been applied in plant cell imaging for the first time. The results reported in this paper may provide a promising route to multiple-color bioimaging using NDs.

Quantitative x-ray fluorescent analysis using fundamental parameters

International Nuclear Information System (INIS)

Sparks, C.J. Jr.

1976-01-01

A monochromatic source of x-rays for sample excitation permits the use of pure elemental standards and relatively simple calculations to convert the measured fluorescent intensities to an absolute basis of weight per unit weight of sample. Only the mass absorption coefficients of the sample for the exciting and the fluorescent radiation need be determined. Besides the direct measurement of these absorption coefficients in the sample, other techniques are considered which require fewer sample manipulations and measurements. These fundamental parameters methods permit quantitative analysis without recourse to the time-consuming process of preparing nearly identical standards

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

International Nuclear Information System (INIS)

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

2003-01-01

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

Properties of alginate fiber spun-dyed with fluorescent pigment dispersion.

Science.gov (United States)

Wang, Ping; Tawiah, Benjamin; Tian, Anli; Wang, Chunxia; Zhang, Liping; Fu, Shaohai

2015-03-15

Spun-dyed alginate fiber was prepared by the spun-dyeing method with the mixture of fluorescent pigment dispersion and sodium alginate fiber spinning solution, and its properties were characterized by SEM, TGA, DSC, and XRD. The results indicate that fluorescent pigment dispersion prepared with esterified poly (styrene-alt maleic acid) had excellent compatibility with sodium alginate fiber spinning solution, and small amount of fluorescent pigment could reduce the viscosity of spun-dyed spinning solutions. SEM photo of spun-dyed alginate fiber indicated that fewer pigment particles deposited on its surface. TGA, DSC, and XRD results suggested that thermal properties and crystal phase of spun-dyed alginate fibers had slight changes compared to the original alginate fibers. The fluorescence intensity of spun-dyed alginate fiber reached its maximum when the content of fluorescent pigment was 4%. The spun-dyed alginate fiber showed excellent rubbing and washing fastness. Copyright © 2014 Elsevier Ltd. All rights reserved.

Fluorescent Nanoparticle Uptake for Brain Tumor Visualization

Directory of Open Access Journals (Sweden)

Rachel Tréhin

2006-04-01

Full Text Available Accurate delineation of tumor margins is vital to the successful surgical resection of brain tumors. We have previously developed a multimodal nanoparticle CLIO-Cy5.5, which is detectable by both magnetic resonance imaging and fluorescence, to assist in intraoperatively visualizing tumor boundaries. Here we examined the accuracy of tumor margin determination of orthotopic tumors implanted in hosts with differing immune responses to the tumor. Using a nonuser-based signal intensity method applied to fluorescent micrographs of 9L gliosarcoma green fluorescent protein (GFP tumors, mean overestimations of 2 and 24 µm were obtained using Cy5.5 fluorescence, compared to the true tumor margin determined by GFP fluorescence, in nude mice and rats, respectively. To resolve which cells internalized the nanoparticle and to quantitate degree of uptake, tumors were disaggregated and cells were analyzed by flow cytometry and fluorescence microscopy. Nanoparticle uptake was seen in both CD11b+ cells (representing activated microglia and macrophages and tumor cells in both animal models by both methods. CD11b+ cells were predominantly found at the tumor margin in both hosts, but were more pronounced at the margin in the rat model. Additional metastatic (CT26 colon and primary (Gli36 glioma brain tumor models likewise demonstrated that the nanoparticle was internalized both by tumor cells and by host cells. Together, these observations suggest that fluorescent nanoparticles provide an accurate method of tumor margin estimation based on a combination of tumor cell and host cell uptake for primary and metastatic tumors in animal model systems and offer potential for clinical translation.

Denaturing of single electrospun fibrinogen fibers studied by deep ultraviolet fluorescence microscopy.

Science.gov (United States)

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.

Copper spherical cavity arrays: Fluorescence enhancement in PFO films

Energy Technology Data Exchange (ETDEWEB)

Spada, Edna R., E-mail: edspada@gmail.com [Instituto de Física de São Carlos, Universidade de São Paulo, Caixa Postal 369, 13560-970, São Carlos, SP (Brazil); Valente, Gustavo T.; Pereira-da-Silva, Marcelo A. [Instituto de Física de São Carlos, Universidade de São Paulo, Caixa Postal 369, 13560-970, São Carlos, SP (Brazil); Sartorelli, Maria L. [Departamento de Física, Universidade Federal de Santa Catarina, Caixa Postal 476, 88040-900, Florianópolis, SC (Brazil); Guimarães, Francisco E.G.; Faria, Roberto M. [Instituto de Física de São Carlos, Universidade de São Paulo, Caixa Postal 369, 13560-970, São Carlos, SP (Brazil)

2017-01-15

This manuscript addresses the use of a well-ordered antidot copper nanostructure as a active substrate for surface enhancement fluorescence (SEF). The antidot array was produced by electrodeposition and nanosphere lithography and characterized by microscopy technique, its successful application as SEF-active substrates was verified using polyfluorene (PFO) as a probe layer. Atomic force microscopy (AFM) was used to evaluate the regularity of the metal surface as well PFO coated process and confocal laser fluorescence microscopy (CLSM) to determine the behavior exhibited by the fluorescent layer due to the existence of the nanostructured surface. No accumulation PFO in the cavities was detected and the more intense emission regions coincides with the position of the cavities and is at about one order of magnitude higher.

Fluorescence Imaging in Genipin Crosslinked Chitosan–Poly(vinyl pyrrolidone Hydrogels

Directory of Open Access Journals (Sweden)

Simon Matcham

2016-10-01

Full Text Available Recent research has identified genipin as a promising natural crosslinking agent for biocompatible hydrogels as genipin is significantly less cytotoxic than current synthetic crosslinking agents, such as glutaraldehyde. Conveniently, fluorophores can be produced when genipin crosslinks. In this study, fluorescence intensity measurements of genipin crosslinked chitosan-poly(vinyl pyrrolidone hydrogels have been explored as a dynamic, in situ method for tracing sol-gel transition. These pH-responsive smart materials have a future in medical applications, in particular in tissue engineering and drug delivery, where methods to follow the process in situ and in real-time are crucial for future advancement. Samples were prepared using deionised water, pH 4, and pH 10 solutions, and studied at 24 and 37 °C over a 24 h period. Both temperature and pH have been found to affect sol-gel transition in the hydrogels studied. The transition from acidic (pH 4 to basic (pH 10 solution resulted in reduced fluorescence intensity suggesting that, under more basic conditions, genipin molecules self-polymerise, reducing the number of molecules available for reaction with the amino groups of chitosan. Three-dimensional representations of the fluorescence present in a hydrogel sample have also been produced from the data, enabling the visualisation of variation in fluorescence with time at the surface of the hydrogel.

Development of a portable system of X-ray fluorescence spectrometry

International Nuclear Information System (INIS)

Mantuano, Andrea; Crisostomo, Jose V.V.; Barros, Mariana J.; Oliveira, Luis F.; Barroso, Regina C.

2009-01-01

This paper develops a compact and portable spectrometry system that will be used at the Laboratory of Applied Physics to the Biomedical and Environmental Sciences of the Institute of Physics/UERJ, Rio de Janeiro, Brazil. The laboratory both prepares the samples and develops the X-ray spectrometry techniques. The techniques of X-ray diffraction and fluorescence on various samples (biological, industrial and environmental) are used, attending to pos-graduation and graduation students, with multidisciplinary characteristics. The Mini-X system consists of X-ray mini tube MINI-X from Amptek with tungsten (W) target, and a compact spectrometer X123, also from Amptek that includes a detector, pre-amplifier, digital pulse processor, and multichannel. All the system is controlled by dedicated microprocessor. This work will present both a methodology for alignment and calibration of the system as far the first measurements performed using the X-ray fluorescence technique on standard samples. The multi elementary analysis by X-ray fluorescence (XRF) is based on the measurements of the characteristic X-ray intensity emitted by the chemical elements components of the samples when excited. Therefore, from the development of this compact and versatile system it will be possible to obtain the fluorescent intensities of the analysed samples at the Laboratory, not only at the research area but at the teaching area. Besides, new laboratory practices are being developed for the discipline of medical physics

X-ray fluorescence hologram data collection with a cooled avalanche photodiode

CERN Document Server

Hayashi, K; Matsubara, E I; Kishimoto, S; Mori, T; Tanaka, M

2002-01-01

A high counting rate X-ray detector with an appropriate energy resolution is desired for high quality X-ray fluorescence hologram measurements because a holographic pattern is detected as extremely small intensity variations of X-ray fluorescence on a large intensity background. A cooled avalanche photodiode (APD), which has about 10% energy resolution and is designed for a high counting rate, fits the above requirements. Reconstructed atomic images from experimental holograms using the APD system provide us a clear view of the first and second neighbor atoms around an emitter. The present result proved that a combination of this APD system and a synchrotron X-ray source enables us to measure a high quality hologram for a reasonable measurement time.

Hyperspectral solar-induced chlorophyll fluorescence of urban tree leaves: Analyses and applications

Science.gov (United States)

Van Wittenberghe, Shari

Solar energy is the primary energy source for life on Earth which is converted into chemical energy through photosynthesis by plants, algae and cyanobacteria, releasing fuel for the organisms' activities. To dissipate excess of absorbed light energy, plants emit chlorophyll (Chl) fluorescence (650-850 nm) from the same location where photosynthesis takes place. Hence, it provides information on the efficiency of primary energy conversion. From this knowledge, many applications on vegetation and crop stress monitoring could be developed, a necessity for our planet under threat of a changing global climate. Even though the Chl fluorescence signal is weak against the intense reflected radiation background, methods for retrieving the solar-induced Chl fluorescence have been refined over the last years, both at leaf and airborne scale. However, a lack of studies on solar-induced Chl fluorescence gives difficulties for the interpretation of the signal. Within this thesis, hyperspectral upward and downward solar-induced Chl fluorescence is measured at leaf level. Fluorescence yield (FY) is calculated as well as different ratios characterizing the emitted Chl fluorescence shape. The research in this PhD dissertation illustrates the influence of several factors on the solar-induced Chl fluorescence signal. For instance, both the intensity of FY and its spectral shape of urban tree leaves are able to change under influence of stress factors such as traffic air pollution. This shows how solar-induced Chl fluorescence could function as an early stress indicator for vegetation. Further, it is shown that the signal contains information on the ultrastructure of the photosynthetic apparatus. Also, it is proven that the leaf anatomical structure and related light scattering properties play a role in the partitioning between upward and downward Chl fluorescence emission. All these findings indicate how the Chl fluorescence spectrum is influenced by factors which also influence

Fluorescence spectroscopic studies on binding of a flavonoid ...

Indian Academy of Sciences (India)

Unknown

six principal binding sites have been identified for several important biomolecules.4 ... lized), tryptophan and quercetin from Sigma were used as received. .... +..... (6). Here Fo and F are the fluorescence intensity from the fluorophore, albumin, at 342 nm in the absence and the presence of different concentrations of ...

Synthesis of novel fluorescent probe Tb(III)-7-carboxymethoxy-4-methylcoumarin complex for sensing of DNA

International Nuclear Information System (INIS)

Hussein, Belal H.M.; Azab, Hassan A.; Fathalla, Walid; Ali, Sherin A.M.

2013-01-01

New fluorescent probe Tb(III) (7-carboxymethoxy-4-methylcoumarin)2(SCN) (C2H5OH)(H2O) was synthesized and characterized by spectroscopy and thermal analysis. The absorption and fluorescence spectra of 7-carboxymethoxy-4-methylcoumarin (CMMC) and Tb(III)–CMMC complex have been measured in different solvents. The interactions of Tb(III)–CMMC complex with calf thymus nucleic acid (CT-DNA) have been investigated using steady state fluorescence measurements. The changes in the fluorescence intensity have been used for the quantitative determination of DNA with LOD of 3.45 ng in methanol–water (9:1, v/v). The association constants of DNA with Tb(III)–CMMC complex was found to be 2.62×1010 M −1 . - Highlights: ► New fluorescent probe Terbium (III)-7-carboxy methoxy-4-methylcoumarin complex has been synthesized and characterized. ► FTIR spectrum of Tb(III)-complex shows a characteristic band for thiocyanate group. ► DNA interaction with Terbium (III)-7-carboxy methoxy-4-methylcoumarin has been studied by fluorescence techniques. ► The change in the fluorescence intensity has been used for the quantitative determination of DNA. ► The result was better than most of the well-known methods including the ethidium bromide method.

COMPARISON OF TWO TEMPERATURE MEASUREMENT METHODS BY UPCONVERSION FLUORESCENCE SPECTRA OF ERBIUM-DOPED LEAD-FLUORIDE NANO-GLASS-CERAMICS

Directory of Open Access Journals (Sweden)

V. A. Aseev

2015-05-01

Full Text Available The study and compare of two temperature measurement methods is performed for the case of a lead-fluoride nano-glassceramics in the range from 317 to 423 K with a view to their application to temperature sensors. A method of temperature measurement by means of violet, green and red upconversion fluorescence spectra regression on latent structures and a method of temperature measurement by two fluorescence bands intensity ratio in green range are considered. It is shown that a four-dimensional space of latent structures is an optimum one in terms of temperature measurement accuracy. It made possible temperature determining with a relative error not larger than 0.15% at temperatures higher than 340 K by making use of fluorescence spectra training set with the step of 10 K. The method using two green bands fluorescence intensity ratio is inferior by the accuracy. Independence of pump power fluctuations is a significant advantage of the second method. To take advantage of the first method a stabilization of the pump power is necessary. The results of the work can be taken into account while developing optical temperature sensors with a better performance (in relation to accuracy and measurement range compared to existing ones which utilize temperature redistribution of fluorescence intensities in two closely-spaced bands or temperature dependence of fluorescence lifetime.

Synthesis and photophysical properties of fluorescence sensing diester-terminated 1,8-naphthalimide

International Nuclear Information System (INIS)

Bojinov, N. I. Georgiev. V. B.

2011-01-01

Full text: The immediate detection and determination of environmental pollutants have been gaining particular importance. In recent years, extensive research has been carried out on fluorescent organic compounds whose photophysical properties are sensitive to environmental changes. Such interest is due to the possibility of tailoring the design of molecular devices for environmental pollution caused by heavy and transition metal ions. Photoinduced electron transfer (PET) using the 'fluorophore-spacer-receptor' format, developed by de Silva, is one of the most popular approaches to the design of fluorescent sensors.; This work reports the synthesis and sensor activity of a 1,8-naphthalimide sensor based on the 'fluorophore-spacer-receptor' format. The diester-terminated 1,8-naphthalimide was found to display sensitive fluorescence signal amplification over a wide pH scale, which has been ascribed to a photoinduced electron transfer from the tertiary amine receptor to the fluorophore. From the changes in the fluorescence intensity, a pKa value of 4.42 was determined, making the synthesized compound of potential use as pH chemosensing material.; In addition, the ability to detect ions has been evaluated in DMF by monitoring the quenching of the fluorescence intensity. Different ions have been tested: Zn 2+ , Ni 2+ , Pb 2+ , Co 2+ , Cu 2+ , and Fe 3+ for this purpose. The results have clearly shown that only Fe 3+ could be efficiently detected

Synthesis, characterization and fluorescence performance of a waterborne polyurethane-based polymeric dye

Energy Technology Data Exchange (ETDEWEB)

Xianhai, Hu, E-mail: hxyh@aiai.edu.cn [CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026 (China); School of Materials and Chemical Engineering, Building Energy Efficiency Research Institute, Anhui University of Architecture, Hefei 230022 (China); Zhang, Xingyuan, E-mail: zxym@ustc.edu.cn [CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026 (China); Liu, Jin [School of Materials and Chemical Engineering, Building Energy Efficiency Research Institute, Anhui University of Architecture, Hefei 230022 (China); Dai, Jiabing [CAS Key Laboratory of Soft Matter Chemistry, Department of Polymer Science and Engineering, University of Science and Technology of China, Hefei 230026 (China)

2013-10-15

A novel anionic waterborne polyurethane-based fluorescent dye WPU-DV26 was synthesized by incorporating the molecular structure of disperse violet 26 (DV26) into the polyurethane chain. The structure of WPU-DV26 was confirmed by means of Fourier transform infrared spectroscopy and UV–vis absorption analysis. Comparing to the UV–vis spectrum of DV26, WPU-DV26 showed a hypsochromic shift from the absorption maxima of 518, 558, 609 nm to 510, 548, 586 nm, respectively. WPU-DV26 can form stable latex in water. The number average molecular weight and its distribution index, and average latex particle size for WPU-DV26 were determined to be 2.33×10{sup 4}, 1.36 and 80 nm, respectively. The improved thermal stability of WPU-DV26 can be attributed to the embedded anthraquinone unit of DV26. It was found that both the intensity and stability of the fluorescence of WPU-DV26 latex were improved significantly compared with those of DV26. -- Highlights: ► A waterborne polyurethane-based polymeric dye was synthesized. ► The fluorescence intensity of WPU-DV26 emulsion was enhanced greatly compared with that of DV26. ► The fluorescence stability of WPU-DV26 emulsion was fine not only for long term storage but also for fluorescence quencher.

Effect of surfactants on the fluorescence spectra of water-soluble ...

Indian Academy of Sciences (India)

TECS

Effect of surfactants on the fluorescence spectra of water-soluble. MEHPPV ... polyacrylic acid (PAA) chains grafted onto their backbone were found to be water soluble, and they exhi- ..... in other words the variation of emission intensity.

Quantitative analysis with energy dispersive X-ray fluorescence analyser

International Nuclear Information System (INIS)

Kataria, S.K.; Kapoor, S.S.; Lal, M.; Rao, B.V.N.

1977-01-01

Quantitative analysis of samples using radioisotope excited energy dispersive x-ray fluorescence system is described. The complete set-up is built around a locally made Si(Li) detector x-ray spectrometer with an energy resolution of 220 eV at 5.94 KeV. The photopeaks observed in the x-ray fluorescence spectra are fitted with a Gaussian function and the intensities of the characteristic x-ray lines are extracted, which in turn are used for calculating the elemental concentrations. The results for a few typical cases are presented. (author)

Solvent-dependent fluorescence enhancement and piezochromism of a carbazole-substituted naphthopyran

Energy Technology Data Exchange (ETDEWEB)

Liu, Lihui; Wang, Aixia [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Wang, Guang, E-mail: wangg923@nenu.edu.cn [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Munyentwari, Alexis [Faculty of Chemistry, Northeast Normal University, Changchun 130024 (China); Zhou, Yihan, E-mail: yhzhou@ciac.ac.cn [National Analytical Research Center of Electrochemistry and Spectroscopy, Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, Changchun 130022 (China)

2015-09-15

A novel carbazole-substituted naphthopyran, 3,3-bis-(4-carbazolylphenyl)-[3H]-naphtho[2,1-b]pyran (CzNP) was designed and synthesized. The new compound exhibited normal photochromism in dichloromethane solution and the UV irradiation did not influence its fluorescence. On the contrary, the fluorescence of CzNP in N,N-dimethylformamide (DMF) was intensively enhanced to 29 times after 60 min of the UV irradiation and this enhanced fluorescence can be quenched by addition of triethylamine (TEA). The study of enhanced extent of fluorescence of CzNP in solvents with different polarities and in mixed solvents demonstrated that the enhanced fluorescence is dependent on the polarity of solvents. The larger the polarity of solvent was, the stronger was the fluorescence of CzNP. CzNP also exhibited piezochromic performance and the pressure led to the cleavage of the C–O bond of pyran ring. - Highlights: • A carbazole-substituted photochromic naphthopyran was designed and synthesized. • The fluorescence was enhanced under the existence of DMF and UV irradiation. • The polarity of solvent was the dominating factor to affect the fluorescence. • The new compound also displayed piezochromic performance.

Preparation of fluorescent polyaniline nanoparticles in aqueous solutions

Energy Technology Data Exchange (ETDEWEB)

Alves, Kleber G. B. [Universidade Federal de Pernambuco, Departamento de Engenharia Mecanica (Brazil); Melo, Etelino F. de [Universidade Federal de Pernambuco, Departamento de Quimica Fundamental (Brazil); Andrade, Cesar A. S. [Universidade Federal de Pernambuco, Departamento de Bioquimica (Brazil); Melo, Celso P. de, E-mail: celso@df.ufpe.br [Universidade Federal de Pernambuco, Departamento de Fisica (Brazil)

2013-01-15

We report the synthesis of stable polyaniline nanoparticles (PANI{sub N}Ps) based on the chemical oxidative polymerization of aniline in aqueous solutions of surfactants. Surfactants of three different types-cationic (dodecyltrimethylammonium bromide-DTAB), anionic (sodium dodecyl sulfate-SDS), and non-ionic (Triton X-405-TX-405)-were used. The resulting PANI{sub N}Ps{sub s}urfactant samples were characterized through UV-Vis, fluorescence and Fourier transform infrared spectroscopies, and scanning electronic microscopy (SEM). We have verified that the color of the PANI{sub N}Ps{sub s}urfactant dispersions is affected by a change in the pH of the solution. The PANI-NPs{sub s}urfactant colloidal suspensions in aqueous solution present a surprising high fluorescence quantum yield value (ranging from 1.9 Multiplication-Sign 10{sup -3} to 6.9 Multiplication-Sign 10{sup -3}) that can be controlled as a function of the pH, a fact that we associate to the corresponding protonation degree of the PANI polymeric chains. We suggest that these fluorescent nanocomposites can find important technological applications in different areas such as organic light emitting devices, biosensors, and pigments for coatings.

Morphing hydrogel patterns by thermo-reversible fluorescence switching.

Science.gov (United States)

Bat, Erhan; Lin, En-Wei; Saxer, Sina; Maynard, Heather D

2014-07-01

Stimuli responsive surfaces that show reversible fluorescence switching behavior in response to temperature changes were fabricated. Oligo(ethylene glycol) methacrylate thermoresponsive polymers with amine end-groups were prepared by atom transfer radical polymerization (ATRP). The polymers were patterned on silicon surfaces by electron beam (e-beam) lithography, followed by conjugation of self-quenching fluorophores. Fluorophore conjugated hydrogel thin films were bright when the gels were swollen; upon temperature-induced collapse of the gels, self-quenching of the fluorophores led to significant attenuation of fluorescence. Importantly, the fluorescence was regained when the temperature was cooled. The fluorescence switching behavior of the hydrogels for up to ten cycles was investigated and the swelling-collapse was verified by atomic force microscopy. Morphing surfaces that change shape several times upon increase in temperature were obtained by patterning multiple stimuli responsive polymers. © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

Optical absorption and fluorescence studies of praseodymium ion in chloroborophosphate glasses

International Nuclear Information System (INIS)

Sharma, Y.K.; Tandon, S.P.

1998-01-01

Full text: The interest in optical absorption and fluorescence studies of rare earth ions in glassy materials is increasing continuously in connection with laser research and related application. The absorption and fluorescence spectra of praseodymium ion in chloroborophosphate glasses have been recorded at room temperature. The chloroborophosphate glass specimens having composition in mob.% Na 2 0 (26.08), B 2 0 3 (14.57), P 2 0 5 (44.85), ZnCl 2 (14.50), Pr 6 0 11 (R) [R= 0.0,0.1 and 0.2 moi.%] have been prepared by melt quenching technique. The spectra consists of seven absorption bands and three fluorescence bands. The observed optical spectra are discussed in terms of energy state and the intensity of the transitions. The various energy interaction parameters like Slater-Condon, Lande', Racah and bonding parameters have been computed. Judd-Ofeit intensity parameters and laser parameters have also been computed. These results shows that praseodymium doped chloroborophosphate glass specimen can be considered as good hosts for laser applications

Fluorescence of Bacteria, Pollens, and Naturally Occurring Airborne Particles: Excitation/Emission Spectra

National Research Council Canada - National Science Library

Hill, Steven C; Mayo, Michael W; Chang, Richard K

2009-01-01

The fluorescence intensity as a function of excitation and emission wavelengths (EEM spectra) was measured for different species of bacteria, biochemical constituents of cells, pollens, and vegetation...

Integrated ultrasonic particle positioning and low excitation light fluorescence imaging

International Nuclear Information System (INIS)

Bernassau, A. L.; Al-Rawhani, M.; Beeley, J.; Cumming, D. R. S.

2013-01-01

A compact hybrid system has been developed to position and detect fluorescent micro-particles by combining a Single Photon Avalanche Diode (SPAD) imager with an acoustic manipulator. The detector comprises a SPAD array, light-emitting diode (LED), lenses, and optical filters. The acoustic device is formed of multiple transducers surrounding an octagonal cavity. By stimulating pairs of transducers simultaneously, an acoustic landscape is created causing fluorescent micro-particles to agglomerate into lines. The fluorescent pattern is excited by a low power LED and detected by the SPAD imager. Our technique combines particle manipulation and visualization in a compact, low power, portable setup

Fluorescence decay time imaging using an imaging photon detector with a radio frequency photon correlation system

Science.gov (United States)

Morgan, Christopher G.; Mitchell, A. C.; Murray, J. G.

1990-05-01

An imaging photon detector has been modified to incorporate fast timing electronics coupled to a custom built photon correlator interfaced to a RISC computer. Using excitation with intensity- muodulated light, fluorescence images can be readily obtained where contrast is determined by the decay time of emission, rather than by intensity. This technology is readily extended to multifrequency phase/demodulation fluorescence imaging or to differential polarised phase fluorometry. The potential use of the correlator for confocal imaging with a laser scanner is also briefly discussed.

Determination of Cerium (IV) Using Rhodamine 6G Fluorescence Quenching

Science.gov (United States)

Zhao, Zh.; Sheng, L.; Su, B.; Tao, C.; Jing, W.

2017-11-01

The interaction between rhodamine 6G (Rh6G) and cerium sulfate was studied by the fluorescence quenching method. In a sulfuric acid medium, the interaction of Ce(IV) with Rh6G results in Rh6G fluorescence quenching. The maximum excitation wavelength (λex) and the maximum emission wavelength (λem) are 530 nm and 555 nm, respectively. A good linearity between the relative fl uorescence intensity (ΔF) and Ce(IV) was observed in the range 0.12-1.08 μg/mL. The detection limit was 1.4 × 10-3 μg/mL. The optimum reaction conditions, influencing factors, and effect of coexisting substances were investigated in the experiment. We found that the concentration of Rh6G was 3.2 × 10-6 mol/L, and the fl uorescence intensity was maximum.

Laser investigation of the non-uniformity of fluorescent species in dental enamel

Science.gov (United States)

Tran, Stephanie U.; Ridge, Jeremy S.; Nelson, Leonard Y.; Seibel, Eric J.

In the present study, artificial type I and type II erosions were created on dental specimen using acetic acid and EDTA respectively. Specimens were prepared by etching extracted teeth samples in acid to varying degrees, after which the absolute fluorescence intensity ratio of the etched enamel relative to sound enamel was recorded for each specimen using 405 and 532 nm laser excitation. Results showed differences in the fluorescence ratio of etched to sound enamel for type I and II erosions. These findings suggest a non-uniform distribution of fluorescent species in the interprismatic region as compared to the prismatic region.

High-quality substrate for fluorescence enhancement using agarose-coated silica opal film.

Science.gov (United States)

Xu, Ming; Li, Juan; Sun, Liguo; Zhao, Yuanjin; Xie, Zhuoying; Lv, Linli; Zhao, Xiangwei; Xiao, Pengfeng; Hu, Jing; Lv, Mei; Gu, Zhongze

2010-08-01

To improve the sensitivity of fluorescence detection in biochip, a new kind of substrates was developed by agarose coating on silica opal film. In this study, silica opal film was fabricated on glass substrate using the vertical deposition technique. It can provide stronger fluorescence signals and thus improve the detection sensitivity. After coating with agarose, the hybrid film could provide a 3D support for immobilizing sample. Comparing with agarose-coated glass substrate, the agarose-coated opal substrates could selectively enhance particular fluorescence signals with high sensitivity when the stop band of the silica opal film in the agarose-coated opal substrate overlapped the fluorescence emission wavelength. A DNA hybridization experiment demonstrated that fluorescence intensity of special type of agarose-coated opal substrates was about four times that of agarose-coated glass substrate. These results indicate that the optimized agarose-coated opal substrate can be used for improving the sensitivity of fluorescence detection with high quality and selectivity.

Fluorescent-Spectroscopic Research of in Vivo Tissues Pathological Conditions

Science.gov (United States)

Giraev, K. M.; Ashurbekov, N. A.; Medzhidov, R. T.

The steady-state spectra of autofluorescence and the reflection coefficient on the excitation wavelength of some stomach tissues in vivo with various pathological conditions (surface gastritis, displasia, cancer) are measured under excitation by the nitrogen laser irradiation (λex=337.1 nm). The contour expansion of obtained fluorescence spectra into contributions of components is conducted by the Gaussian-Lorentzian curves method. It is shown that at least 7 groups of fluorophores forming a total luminescence spectrum can be distinguished during the development of displasia and tumor processes. The correlation of intensities of flavins and NAD(P)·H fluorescence is determined and the degree of respiratory activity of cells for the functional condition considered is estimated. The evaluations of the fluorescence quantum yield of the tissue's researched are given.

Fundamental parameters method for quantitative energy dispersive x-ray fluorescence analysis

International Nuclear Information System (INIS)

Demirel, H.; Zararsiz, A.

1986-01-01

In this study, the requirement of the standart material in photon excited energy distributed X-ray fluorescence analysis has been removed. The interaction of X-rays with matter has been taken into account. A computer program has been developed by using the fundamental parameters of X-ray fluorescence technique and the spectral intensity 'K' of pure elements at saturation thickness has been obtained. For experimental purpose a convenient source-target-detector geometry has been designed. In order to excite the samples,Cd-109 radioisotope source has been used. The peak intensities has been obtained in a vacum chamber by counting the emitted X-rays. The calculation of concentration has been performed for double mixed samples correcting the effects of absorption and enchancement factors. The results were in conformity with their certificate values. (author)

Tools for the quantitative analysis of sedimentation boundaries detected by fluorescence optical analytical ultracentrifugation.

Directory of Open Access Journals (Sweden)

Huaying Zhao

Full Text Available Fluorescence optical detection in sedimentation velocity analytical ultracentrifugation allows the study of macromolecules at nanomolar concentrations and below. This has significant promise, for example, for the study of systems of high-affinity protein interactions. Here we describe adaptations of the direct boundary modeling analysis approach implemented in the software SEDFIT that were developed to accommodate unique characteristics of the confocal fluorescence detection system. These include spatial gradients of signal intensity due to scanner movements out of the plane of rotation, temporal intensity drifts due to instability of the laser and fluorophores, and masking of the finite excitation and detection cone by the sample holder. In an extensive series of experiments with enhanced green fluorescent protein ranging from low nanomolar to low micromolar concentrations, we show that the experimental data provide sufficient information to determine the parameters required for first-order approximation of the impact of these effects on the recorded data. Systematic deviations of fluorescence optical sedimentation velocity data analyzed using conventional sedimentation models developed for absorbance and interference optics are largely removed after these adaptations, resulting in excellent fits that highlight the high precision of fluorescence sedimentation velocity data, thus allowing a more detailed quantitative interpretation of the signal boundaries that is otherwise not possible for this system.

Theory of analytical curves in atomic fluorescence flame spectrometry

NARCIS (Netherlands)

Hooymayers, H.P.

An explicit expression for the intensity of atomic resonance fluorescence as a function of atomic concentration in a flame is derived under certain idealized conditions. The expression is generally valid for a pure Doppler absorption line profile as well as for a combined Doppler and collisional

Fluorescence spectroscopy of conformational changes of single LH2 complexes

NARCIS (Netherlands)

Rutkauskas, D.; Novoderezhkin, V.; Cogdell, R.J.; van Grondelle, R.

2005-01-01

We have investigated the energy landscape of the bacterial photosynthetic peripheral light-harvesting complex LH2 of purple bacterium Rhodopseudomonas acidophila by monitoring sequences of fluorescence spectra of single LH2 assemblies, at room temperature, with different excitation intensities as

Intrinsic fluorescence of protein in turbid media using empirical relation based on Monte Carlo lookup table

Science.gov (United States)

Einstein, Gnanatheepam; Udayakumar, Kanniyappan; Aruna, Prakasarao; Ganesan, Singaravelu

2017-03-01

Fluorescence of Protein has been widely used in diagnostic oncology for characterizing cellular metabolism. However, the intensity of fluorescence emission is affected due to the absorbers and scatterers in tissue, which may lead to error in estimating exact protein content in tissue. Extraction of intrinsic fluorescence from measured fluorescence has been achieved by different methods. Among them, Monte Carlo based method yields the highest accuracy for extracting intrinsic fluorescence. In this work, we have attempted to generate a lookup table for Monte Carlo simulation of fluorescence emission by protein. Furthermore, we fitted the generated lookup table using an empirical relation. The empirical relation between measured and intrinsic fluorescence is validated using tissue phantom experiments. The proposed relation can be used for estimating intrinsic fluorescence of protein for real-time diagnostic applications and thereby improving the clinical interpretation of fluorescence spectroscopic data.

Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

International Nuclear Information System (INIS)

Gartia, Manas Ranjan; Hsiao, Austin; Logan Liu, G; Sivaguru, Mayandi; Chen Yi

2011-01-01

We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

Energy Technology Data Exchange (ETDEWEB)

Gartia, Manas Ranjan [Department of Nuclear, Plasma and Radiological Engineering, University of Illinois, Urbana, IL 61801 (United States); Hsiao, Austin; Logan Liu, G [Department of Bioengineering, University of Illinois, Urbana, IL 61801 (United States); Sivaguru, Mayandi [Institute for Genomic Biology, University of Illinois, Urbana, IL 61801 (United States); Chen Yi, E-mail: loganliu@illinois.edu [Department of Electrical and Computer Engineering, University of Illinois, Urbana, IL 61801 (United States)

2011-09-07

We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

Segmentation and classification of cell cycle phases in fluorescence imaging.

Science.gov (United States)

Ersoy, Ilker; Bunyak, Filiz; Chagin, Vadim; Cardoso, M Christina; Palaniappan, Kannappan

2009-01-01

Current chemical biology methods for studying spatiotemporal correlation between biochemical networks and cell cycle phase progression in live-cells typically use fluorescence-based imaging of fusion proteins. Stable cell lines expressing fluorescently tagged protein GFP-PCNA produce rich, dynamically varying sub-cellular foci patterns characterizing the cell cycle phases, including the progress during the S-phase. Variable fluorescence patterns, drastic changes in SNR, shape and position changes and abundance of touching cells require sophisticated algorithms for reliable automatic segmentation and cell cycle classification. We extend the recently proposed graph partitioning active contours (GPAC) for fluorescence-based nucleus segmentation using regional density functions and dramatically improve its efficiency, making it scalable for high content microscopy imaging. We utilize surface shape properties of GFP-PCNA intensity field to obtain descriptors of foci patterns and perform automated cell cycle phase classification, and give quantitative performance by comparing our results to manually labeled data.

Control of the blue fluorescent protein with advanced evolutionary pulse shaping

International Nuclear Information System (INIS)

Tkaczyk, Eric R.; Mauring, Koit; Tkaczyk, Alan H.; Krasnenko, Veera; Ye, Jing Yong; Baker, James R.; Norris, Theodore B.

2008-01-01

We demonstrate optical coherent control of the two-photon fluorescence of the blue fluorescent protein (BFP), which is of interest in investigations of protein-protein interactions. In addition to biological relevance, BFP represents an interesting target for coherent control from a chemical perspective due to its many components of highly nonexponential fluorescence decay and low quantum yield resulting from excited state isomerization. Using a genetic algorithm with a multiplicative (rather than ratiometric) fitness parameter, we are able to control the ratio of BFP fluorescence to second-harmonic generation without a considerable drop in the maximized signal. The importance of linear chirp and power-scaling on the discrimination process is investigated in detail

Quantum electrodynamics of the internal source x-ray holographies: Bremsstrahlung, fluorescence, and multiple-energy x-ray holography

International Nuclear Information System (INIS)

Miller, G.A.; Sorensen, L.B.

1997-01-01

Quantum electrodynamics (QED) is used to derive the differential cross sections measured in the three new experimental internal source ensemble x-ray holographies: bremsstrahlung (BXH), fluorescence (XFH), and multiple-energy (MEXH) x-ray holography. The polarization dependence of the BXH cross section is also obtained. For BXH, we study analytically and numerically the possible effects of the virtual photons and electrons which enter QED calculations in summing over the intermediate states. For the low photon and electron energies used in the current experiments, we show that the virtual intermediate states produce only very small effects. This is because the uncertainty principle limits the distance that the virtual particles can propagate to be much shorter than the separation between the regions of high electron density in the adjacent atoms. We also find that using the asymptotic form of the scattering wave function causes about a 5 10% error for near forward scattering. copyright 1997 The American Physical Society

Increased signal intensities in the dentate nucleus and globus pallidus on unenhanced T1-weighted images: evidence in children undergoing multiple gadolinium MRI exams

Energy Technology Data Exchange (ETDEWEB)

Hu, Houchun H.; Pokorney, Amber; Towbin, Richard B.; Miller, Jeffrey H. [Phoenix Children' s Hospital, Department of Medical Imaging and Radiology, Phoenix, AZ (United States)

2016-10-15

Recent reports have suggested residual gadolinium deposition in the brain in subjects undergoing multiple contrast-enhanced MRI exams. These findings have raised some concerns regarding gadolinium-based contrast agent (GBCA) usage and retention in brain tissues. To summarize findings of hyperintense brain structures on precontrast T1-weighted images in 21 children undergoing multiple GBCA MRI exams. This retrospective study involved 21 patients, each of whom received multiple MRI examinations (range: 5-37 exams) with GBCA over the course of their medical treatment (duration from first to most recent exam: 1.2-12.9 years). The patients were between 0.9 and 14.4 years of age at the time of their first GBCA exam. Regions of interest were drawn in the dentate nucleus and the globus pallidus on 2-D fast spin echo images acquired at 1.5 T. The signal intensities of these two structures were normalized by that of the corpus callosum genu. Signal intensity ratios from these patients were compared to control patients of similar ages who have never received GBCA. Signal intensity ratios increased between the first and the most recent MRI exam in all 21 patients receiving GBCA, with an increase of 18.6%±12.7% (range: 0.5% to 47.5%) for the dentate nucleus and 12.4%±7.4% (range: -1.2% to 33.7%) for the globus pallidus (P<0.0001). Signal intensity ratios were also higher in GBCA patients than in controls (P<0.01). The degree of signal intensity enhancement did not correlate with statistical significance to the cumulative number or volume of GBCA administrations each patient received, the patient's age or the elapsed time between the first and most recent GBCA MRI exams. These results in children are consistent with recent findings in adults, suggesting possible gadolinium deposition in the brain. (orig.)

The sensitivity and selectivity properties of a fluorescence sensor based on quinoline-Bodipy

Energy Technology Data Exchange (ETDEWEB)

Nuri Kursunlu, Ahmed, E-mail: ankursunlu@gmail.com; Guler, Ersin

2014-01-15

A novel florescence sensor (Q-BODIPY) based on quinoline-Bodipy (quinoline-boradiazaindacene) was prepared by ‘click chemistry’ in several stages. The sensing actions of Q-BODIPY were confirmed by UV–vis titration, emission and excitation spectroscopic studies in presence of Mn{sup 2+}, Co{sup 2+}, Ni{sup 2+}, Cu{sup 2+}, Zn{sup 2+}, Cd{sup 2+}, Sn{sup 2+}, Hg{sup 2+}, Pb{sup 2+}, La{sup 3+}, Ga{sup 3+}, Er{sup 3+} and Yb{sup 3+} ions in methanol:H{sub 2}O (1:1) medium. Whereas some metal ions can only cause quenching effect on the fluorescence intensity of Q-BODIPY, some of them show an increase in fluorescence intensity. The stoichiometry of host–guest complexes formed was determined by Job′s plot method. The binding constants were calculated by Stern–Volmer method. As a fluorescence sensor, Q-BODIPY shows the best selectivity performance against Zn{sup 2+} ions in according to all spectroscopic data. -- Highlights: • Q-BODIPY prepared by several techniques shows a fluorescent behavior toward p, d and f block metal ions. • Q-BODIPY has both a more sensitivity and more effective ability for the detection of Zn(II) ion. • The synthesis strategies to produce Bodipy′s with metal coordinating offer a new approach for the design of novel fluorescence sensors.

Spectral ’Fingerprinting’ of Phytoplankton Populations by Two-Dimensional Fluorescence and Fourier-Transform-Based Pattern Recognition.

Science.gov (United States)

1985-07-08

Chlorella vulgaris and Aaphidinium carterae ) were synthetically added to each other in varying proportions from 0% C. vulgaris fluorescence to 100...C. vulgaris fluorescence in 10% increments. The proportions were calculated based on the maximum fluorescence intensity. The series of mixtures was...then correlated against the original 9 standard library in which the C. vulgaris and A. Carterae REs were represented. Table IV presents the results of

Detection of intaoral lesions using a fluorescence camera

Science.gov (United States)

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.

Fluorescent carbon dots nanosensor for label-free determination of vitamin B12 based on inner filter effect

Science.gov (United States)

Ding, Longhua; Yang, Hongmei; Ge, Shenguang; Yu, Jinghua

2018-03-01

A simple and effective fluorescent assay for the determination of vitamin B12 was developed. In this study, carbon dots (CDs) were prepared by one-pot hydrothermal method and directly used as a fluorophore in the inner filter effect (IFE). Both of the maximum absorption peak of vitamin B12 and excitation maxima of CDs are located at 360 nm, hence, the excited light of CDs can be absorbed by vitamin B12, resulting in the fluorescence reduction of CDs. And the fluorescence intensity of CDs decreases with the increasing concentration of vitamin B12. This IFE-based sensing strategy shows a good linear relationship between the normalized fluorescence intensity and the concentration of vitamin B12 ranging from 0 to 60 μM, with a limit of detection (LOD) of 0.1 μM at a signal-to-noise ratio of 3. Furthermore, this proposed approach was successfully applied to vitamin B12 sensing in injections. This IFE sensing platform based on various fluorescent nanomaterials has a high promise for the detection of other biomolecules due to its inherent convenience.

Fluorescent cadmium sulfide nanoparticles for selective and sensitive detection of toxic pesticides in aqueous medium

International Nuclear Information System (INIS)

Walia, Shanka; Acharya, Amitabha

2014-01-01

The detection of pesticide residues in ground water, food, or soil samples is extremely important. The currently available laboratory techniques have several drawbacks and needs to be replaced. Fluorescent chemosensors for pesticide detection were reported in the literature, with few reports published on quantum dot-based pesticide sensors, but none of these were focused toward differentiating organophosphorus and organochlorine pesticides specifically. In this respect, glutathione-coated CdS nanoparticles were synthesized and characterized. The TEM studies of the nanoparticles suggested mostly monodispersed spherical particles, with size in the range of 11.5±1 nm. The prepared fluorescent nanoparticles were found to selectively recognize organochlorine pesticide dicofol among all the other pesticides studied, by increasing the fluorescence intensity of the nanoparticles ∼ 2.5 times. Similar studies carried out with organophosphorous pesticide dimethoate did not result any change in the fluorescence intensity of the nanoparticles. Further studies carried out with commercially available pesticide solutions, also confirmed similar results. The TEM, SEM, and DLS studies suggested aggregation of the nanoparticles in the presence of dicofol. Control experiments suggested possible role of both amine and carboxylic acid functional groups of glutathione in the recognition of dicofol. The limit of detection of dicofol was found to be ∼ 55±11 ppb.Graphical AbstractGlutathione-coated CdS nanoparticles selectively recognize organochlorine pesticide dicofol among all the other pesticides studied, by increasing the fluorescence intensity of the nanoparticles. The TEM, SEM, and DLS studies suggested aggregation of the nanoparticles in the presence of dicofol

Fluorescent cadmium sulfide nanoparticles for selective and sensitive detection of toxic pesticides in aqueous medium

Energy Technology Data Exchange (ETDEWEB)

Walia, Shanka; Acharya, Amitabha, E-mail: amitabhachem@gmail.com [CSIR-Institute of Himalayan Bioresource Technology, Biotechnology Division (India)

2014-12-15

The detection of pesticide residues in ground water, food, or soil samples is extremely important. The currently available laboratory techniques have several drawbacks and needs to be replaced. Fluorescent chemosensors for pesticide detection were reported in the literature, with few reports published on quantum dot-based pesticide sensors, but none of these were focused toward differentiating organophosphorus and organochlorine pesticides specifically. In this respect, glutathione-coated CdS nanoparticles were synthesized and characterized. The TEM studies of the nanoparticles suggested mostly monodispersed spherical particles, with size in the range of 11.5±1 nm. The prepared fluorescent nanoparticles were found to selectively recognize organochlorine pesticide dicofol among all the other pesticides studied, by increasing the fluorescence intensity of the nanoparticles ∼ 2.5 times. Similar studies carried out with organophosphorous pesticide dimethoate did not result any change in the fluorescence intensity of the nanoparticles. Further studies carried out with commercially available pesticide solutions, also confirmed similar results. The TEM, SEM, and DLS studies suggested aggregation of the nanoparticles in the presence of dicofol. Control experiments suggested possible role of both amine and carboxylic acid functional groups of glutathione in the recognition of dicofol. The limit of detection of dicofol was found to be ∼ 55±11 ppb.Graphical AbstractGlutathione-coated CdS nanoparticles selectively recognize organochlorine pesticide dicofol among all the other pesticides studied, by increasing the fluorescence intensity of the nanoparticles. The TEM, SEM, and DLS studies suggested aggregation of the nanoparticles in the presence of dicofol.

Glycine Insertion Makes Yellow Fluorescent Protein Sensitive to Hydrostatic Pressure

Science.gov (United States)

Watanabe, Tomonobu M.; Imada, Katsumi; Yoshizawa, Keiko; Nishiyama, Masayoshi; Kato, Chiaki; Abe, Fumiyoshi; Morikawa, Takamitsu J.; Kinoshita, Miki; Fujita, Hideaki; Yanagida, Toshio

2013-01-01

Fluorescent protein-based indicators for intracellular environment conditions such as pH and ion concentrations are commonly used to study the status and dynamics of living cells. Despite being an important factor in many biological processes, the development of an indicator for the physicochemical state of water, such as pressure, viscosity and temperature, however, has been neglected. We here found a novel mutation that dramatically enhances the pressure dependency of the yellow fluorescent protein (YFP) by inserting several glycines into it. The crystal structure of the mutant showed that the tyrosine near the chromophore flipped toward the outside of the β-can structure, resulting in the entry of a few water molecules near the chromophore. In response to changes in hydrostatic pressure, a spectrum shift and an intensity change of the fluorescence were observed. By measuring the fluorescence of the YFP mutant, we succeeded in measuring the intracellular pressure change in living cell. This study shows a new strategy of design to engineer fluorescent protein indicators to sense hydrostatic pressure. PMID:24014139

Time resolved fluorescence of cow and goat milk powder

Science.gov (United States)

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

2017-01-01

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

Optical quantification of caries-like lesions in vitro by use of a fluorescent dye

International Nuclear Information System (INIS)

Van de Rijke, J.W.; Ten Bosch, J.J.

1990-01-01

An experimental method was developed for measurement of the fluorescence intensity of a dye that was introduced into caries-like lesions in vitro. A distinct pattern of change of fluorescence intensity with time appeared, displaying a plateau value and a peak value for each measurement. Both plateau and peak values showed a linear correlation with calcium loss, as measured with longitudinal microradiography. The correlation coefficients were r = 0.87 for plateau values and r = 0.89 for peak values. The difference in scattering by dry and wet caries lesions was also measured with the same equipment, which showed a linear correlation with calcium loss of r = -0.53

Giemsa as a fluorescent dye for mineralizing bone-like nodules in vitro

International Nuclear Information System (INIS)

Querido, W; Farina, M; Balduino, A

2012-01-01

Giemsa was first used as a fluorescent dye for mineralized bone and cartilage in tissue sections. The aim of this study was to establish the use of Giemsa as a fluorescent dye for mineralizing bone-like nodules produced in cell cultures. Osteoblasts were grown under mineralizing conditions for 14 days, producing typical bone-like nodules. Upon staining with Giemsa stock solution for 1 min, the mineralizing nodules could be selectively visualized emitting intense green and red fluorescence when observed under blue and green illumination, respectively. The textural details of the nodules were clearly observed under fluorescence microscopy, allowing to identify regions with different degrees of mineralization. The mineralized nature of the nodules was confirmed using von Kossa's method, Alizarin Red S staining and x-ray mapping for Ca and P in a scanning electron microscope, showing a strong correlation between the mineralizing and the fluorescent nodules. The selective fluorescence was related to the mineral phase, being absent in decalcified samples. The use of Giemsa as a fluorescent dye for mineralizing bone-like nodules presents a simple alternative method to quickly analyze biomineralization assays in vitro under fluorescence microscopy, particularly in the biological evaluation of biomaterials. (communication)

Enhancement of single-molecule fluorescence signals by colloidal silver nanoparticles in studies of protein translation.

Science.gov (United States)

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.

Enhancement of Single Molecule Fluorescence Signals by Colloidal Silver Nanoparticles in Studies of Protein Translation

Science.gov (United States)

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

Detection of ultra-low oxygen concentration based on the fluorescence blinking dynamics of single molecules

Science.gov (United States)

Wu, Ruixiang; Chen, Ruiyun; Zhou, Haitao; Qin, Yaqiang; Zhang, Guofeng; Qin, Chengbing; Gao, Yan; Gao, Yajun; Xiao, Liantuan; Jia, Suotang

2018-01-01

We present a sensitive method for detection of ultra-low oxygen concentrations based on the fluorescence blinking dynamics of single molecules. The relationship between the oxygen concentration and the fraction of time spent in the off-state, stemming from the population and depopulation of triplet states and radical cationic states, can be fitted with a two-site quenching model in the Stern-Volmer plot. The oxygen sensitivity is up to 43.42 kPa-1 in the oxygen partial pressure region as low as 0.01-0.25 kPa, which is seven times higher than that of the fluorescence intensity indicator. This method avoids the limitation of the sharp and non-ignorable fluctuations that occur during the measurement of fluorescence intensity, providing potential applications in the field of low oxygen-concentration monitoring in life science and industry.

Microplate-compatible total internal reflection fluorescence microscopy for receptor pharmacology

Science.gov (United States)

Chen, Minghan; Zaytseva, Natalya V.; Wu, Qi; Li, Min; Fang, Ye

2013-05-01

We report the use of total internal reflection fluorescence (TIRF) microscopy for analyzing receptor pharmacology and the development of a microplate-compatible TIRF imaging system. Using stably expressed green fluorescence protein tagged β2-adrenergic receptor as the reporter, we found that the activation of different receptors results in distinct kinetic signatures of the TIRF intensity of cells. These TIRF signatures closely resemble the characteristics of their respective label-free dynamic mass redistribution signals in the same cells. This suggests that TIRF in microplate can be used for profiling and screening drugs.

Study on serum fluorescence spectra based on wavelet transform

African Journals Online (AJOL)

STORAGESEVER

2010-02-08

Feb 8, 2010 ... Blood plays an important role in clinical diagnosis and treatment and as such, the analysis of blood spectrum will be of very important practical significance. Serum fluorescence emission intensity is closely related with the excitation wavelength; when the excitation wavelength is 230 nm, the blood.

Study on serum fluorescence spectra based on wavelet transform ...

African Journals Online (AJOL)

Blood plays an important role in clinical diagnosis and treatment and as such, the analysis of blood spectrum will be of very important practical significance. Serum fluorescence emission intensity is closely related with the excitation wavelength; when the excitation wavelength is 230 nm, the blood lipid concentration and ...

Fluorescence-based detection of nitric oxide in aqueous and methanol media using a copper(II) complex.

Science.gov (United States)

Mondal, Biplab; Kumar, Pankaj; Ghosh, Pokhraj; Kalita, Apurba

2011-03-14

The quenched fluorescent intensity of a copper(II) complex, 1, of a fluorescent ligand, in degassed methanol or aqueous (buffered at pH 7.2) solution, was found to reappear on exposure to nitric oxide. Thus, it can function as a fluorescence based nitric oxide sensor. It has been found that the present complex can be used to sense nanomolar quantities of nitric oxide in both methanol and pH 7.2 buffered-water medium.

Effect of powder sample granularity on fluorescent intensity and on thermal parameters in x-ray diffraction Rietveld analysis

International Nuclear Information System (INIS)

Sparks, C.J.; Specht, E.D.; Ice, G.E.; Kumar, R.; Zschack, P.; Shiraishi, T.; Hisatsune, K.

1991-01-01

The effect of sample granularity on diffracted x-ray intensity was evaluated by measuring the 2θ dependence of x-ray fluorescence from various samples. Measurements were made in the symmetric geometry on samples ranging from single crystals to highly absorbing coarse powders. A characteristic shape for the absorption correction was observed. A demonstration of the sensitivity of Rietveld refined site occupation parameters is made on CuAu and Cu 50 Au 44 Ni 6 alloys refined with and without granularity corrections. These alloys provide a good example of the effect of granularity due to their large linear x-ray absorption coefficients. Sample granularity and refined thermal parameters obtained from the Rietveld analysis were found to be correlated. Without a granularity correction, the refined thermal parameters are too low and can actually become negative in an attempt to compensate for granularity. A general shape for granularity correction can be included in refinement procedures. If no granularity correction is included, data should be restricted to above 30 degrees 2θ, and thermal parameters should be ignored unless extreme precautions are taken to produce >5 μm particles and high packing densities

DAF-fluorescence without NO: elicitor treated tobacco cells produce fluorescing DAF-derivatives not related to DAF-2 triazol.

Science.gov (United States)

Rümer, Stefan; Krischke, Markus; Fekete, Agnes; Mueller, Martin J; Kaiser, Werner M

2012-08-15

Diaminofluorescein-dyes (DAFs) are widely used for visualizing NO· production in biological systems. Here it was examined whether DAF-fluorescence could be evoked by other means than nitrosation. Tobacco (Nicotiana tabacum) suspension cells treated with the fungal elicitor cryptogein released compound(s) which gave a fluorescence increase in the cell-free filtrate after addition of DAF-2 or DAF-FM or DAR-4M. DAF-reactive compounds were relatively stable and identified as reaction products of H(2)O(2) plus apoplastic peroxidase (PO). CPTIO prevented formation of these products. Horseradish-peroxidase (HR-PO) plus H(2)O(2) also generated DAF-fluorescence in vitro. Using RP-HPLC with fluorescence detection, DAF derivatives were further analyzed. In filtrates from cryptogein-treated cells, fluorescence originated from two novel DAF-derivatives also obtained in vitro with DAF-2+HR-PO+H(2)O(2). DAF-2T was only detected when an NO donor (DEA-NO) was present. Using high resolution mass spectrometry, the two above-described novel DAF-reaction products were tentatively identified as dimers. In cells preloaded with DAF-2 DA and incubated with or without cryptogein, DAF-fluorescence originated from a complex pattern of multiple products different from those obtained in vitro. One specific peak was responsive to exogenous H(2)O(2), and another, minor peak eluted at or close to DAF-2T. Thus, in contrast to the prevailing opinion, DAF-2 can be enzymatically converted into a variety of highly fluorescing derivatives, both inside and outside cells, of which none (outside) or only a minor part (inside) appeared NO· dependent. Accordingly, DAF-fluorescence and its prevention by cPTIO do not necessarily indicate NO· production. Copyright © 2012 Elsevier Inc. All rights reserved.

Measurement of fluorescence emission spectrum of few strongly driven atoms using an optical nanofiber.

Science.gov (United States)

Das, Manoj; Shirasaki, A; Nayak, K P; Morinaga, M; Le Kien, Fam; Hakuta, K

2010-08-02

We show that the fluorescence emission spectrum of few atoms can be measured by using an optical nanofiber combined with the optical heterodyne and photon correlation spectroscopy. The observed fluorescence spectrum of the atoms near the nanofiber shows negligible effects of the atom-surface interaction and agrees well with the Mollow triplet spectrum of free-space atoms at high excitation intensity.

A novel and sensitive fluorescence sensor for glutathione detection by controlling the surface passivation degree of carbon quantum dots.

Science.gov (United States)

Pan, Jiahong; Zheng, Zengyao; Yang, Jianying; Wu, Yaoyu; Lu, Fushen; Chen, Yaowen; Gao, Wenhua

2017-05-01

A novel fluorescence sensor based on controlling the surface passivation degree of carbon quantum dots (CQDs) was developed for glutathione (GSH) detection. First, we found that the fluorescence intensity of the CQDs which was obtained by directly pyrolyzing citric acid would increased largely after the surface passivation treatment by 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide (EDC). In the light of this phenomenon, we designed a simple, rapid and selective fluorescence sensor based on the surface passivated CQDs. A certain and excess amount of EDC were mixed with GSH, part of EDC would form a stable complex with GSH owing to the exposed sulfhydryl group of GSH. As the synthesized CQDs were added into the above mixture solution, the fluorescence intensity of the (EDC/GSH)/CQDs mixture solution could be directly related to the amount of GSH. Compared to other fluorescence analytical methods, the fluorescence sensor we design is neither the traditional fluorescent "turn on" probes nor "turn off" probes. It is a new fluorescence analytical method that target object indirectly control the surface passivation degree of CQDs so that it can realize the detection of the target object. Moreover, the proposed method manifested great advantages including short analysis time, low cost and ease of operation. Copyright © 2017 Elsevier B.V. All rights reserved.