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Sample records for fluorescence microprobe sensitivity

  1. Development and applications of an epifluorescence module for synchrotron x-ray fluorescence microprobe imaging

    SciTech Connect

    Miller, Lisa M.; Smith, Randy J.; Ruppel, Meghan E.; Ott, Cassandra H.; Lanzirotti, Antonio

    2005-06-15

    Synchrotron x-ray fluorescence (XRF) microprobe is a valuable analysis tool for imaging trace element composition in situ at a resolution of a few microns. Frequently, epifluorescence microscopy is beneficial for identifying the region of interest. To date, combining epifluorescence microscopy with x-ray microprobe has involved analyses with two different microscopes. We report the development of an epifluorescence module that is integrated into a synchrotron XRF microprobe beamline, such that visible fluorescence from a sample can be viewed while collecting x-ray microprobe images simultaneously. This unique combination has been used to identify metal accumulation in Alzheimer's disease plaques and the mineral distribution in geological samples. The flexibility of this accessory permits its use on almost any synchrotron x-ray fluorescence microprobe beamline and applications in many fields of science can benefit from this technology.

  2. A wavelength dispersive detector for synchrotron x-ray fluorescence microprobe analysis (abstract)

    NASA Astrophysics Data System (ADS)

    Rivers, Mark L.; Sutton, Stephen R.

    1995-02-01

    The synchrotron x-ray fluorescence (SXRF) microprobe has proven to be a valuable tool for trace element research. It permits analysis down to a few parts per million of many elements in a spot size of less than 10 μm. Existing SXRF microprobes are using energy dispersive detectors (EDS), either Si(Li) or intrinsic Ge diodes. Such detectors have the advantage of collecting the entire fluorescence spectrum at once. They can also be positioned to collect a relatively large solid angle. However, EDS detectors suffer from several significant problems: resolution at Fe Kα is about 150 eV, which is roughly 60 times the natural linewidth; the maximum count rate is less than 20 000 counts/s in the entire spectrum; there is significant low-energy background due to scattering and incomplete charge collection in the device. For geochemical analyses these limitations preclude trace element analyses in the presence of a large amount of a high atomic number element: for example, trace element studies of galena (PbS) and zircon (ZrSiO4), or measurements of Cr or Ti in minerals with more than a few percent Fe or Mn. The poor energy resolution prevents the measurement of small amounts of rare-earth elements in samples with significant concentrations of first-row transition elements. Wavelength dispersive spectrometers, based upon Bragg diffraction from a bent crystal, have several distinct advantages over EDS detectors. The resolution at Fe Kα is about 10 eV, or only 4 times the natural linewidth. This permits the analysis of rare-earth elements and also lowers the background which improves detection limits to the 0.1 ppm range. The WDS spectrometer only detects a single energy at once, so it is possible to measure trace elements in the presence of intense fluorescence of a major element. We have installed a commercial wavelength dispersive spectrometer (model WDX-3PC from Microspec Corp., Fremont, CA) on the X-26A microprobe beamline at the NSLS. The spectrometer can scan the

  3. A High-Speed Detector System for X-ray Fluorescence Microprobes.

    SciTech Connect

    Siddons,P.D.; Dragone, A.; De Geronimo, g.; Kuczewski, A.; Kuczewski, J.; O

    2006-10-29

    We have developed a high-speed system for collecting x-ray fluorescence microprobe data, based on ASICs developed at BNL and high-speed processors developed by CSIRO. The system can collect fluorescence data in a continuous raster scan mode, and present elemental images in real time using Ryan's Dynamic Analysis algorithm. We will present results from a 32-element prototype array illustrating the concept. The final instrument will have 384 elements arranged in a square array around a central hole.

  4. A hard x-ray scanning microprobe for fluorescence imaging and microdiffraction at the Advanced Photon Source

    SciTech Connect

    Cai, L.; Lai, B.; Yun, W.; Ilinski, P.; Legnini, D.; Maser, J.; Rodrigues, W.

    1999-11-02

    A hard x-ray scanning microprobe based on zone plate optics and undulator radiation, in the energy region from 6 to 20 keV, has reached a focal spot size (FWHM) of 0.15 {micro}m (v) x 0.6 {micro}m (h), and a photon flux of 4 x 10{sup 9} photons/sec/0.01%BW. Using a slit 44 meters upstream to create a virtual source, a circular beam spot of 0.15 {micro}m in diameter can be obtained with a photon flux of one order of magnitude less. During fluorescence mapping of trace elements in a single human ovarian cell, the microprobe exhibited an imaging sensitivity for Pt (L{sub a} line) of 80 attograms/{micro}m{sup 2} for a count rate of 10 counts per second. The x-ray microprobe has been used to map crystallographic strain and multiquantum well thickness in micro-optoelectronic devices produced with the selective area growth technique.

  5. Using Synchrotron X-ray Fluorescence Microprobes in the Study of Metal Homeostasis in Plants

    SciTech Connect

    Punshon, T.; Guerinot, M; Lanzirotti, A

    2009-01-01

    Background and Aims: This Botanical Briefing reviews the application of synchrotron X-ray fluorescence (SXRF) microprobes to the plant sciences; how the technique has expanded our knowledge of metal(loid) homeostasis, and how it can be used in the future. Scope: The use of SXRF microspectroscopy and microtomography in research on metal homeostasis in plants is reviewed. The potential use of SXRF as part of the ionomics toolbox, where it is able to provide fundamental information on the way that plants control metal homeostasis, is recommended. Conclusions: SXRF is one of the few techniques capable of providing spatially resolved in-vivo metal abundance data on a sub-micrometre scale, without the need for chemical fixation, coating, drying or even sectioning of samples. This gives researchers the ability to uncover mechanisms of plant metal homeostasis that can potentially be obscured by the artefacts of sample preparation. Further, new generation synchrotrons with smaller beam sizes and more sensitive detection systems will allow for the imaging of metal distribution within single living plant cells. Even greater advances in our understanding of metal homeostasis in plants can be gained by overcoming some of the practical boundaries that exist in the use of SXRF analysis.

  6. Quantifying trace elements in individual aquatic protist cells with a synchrotron x-ray fluorescence microprobe.

    SciTech Connect

    Twining, B. S.; Baines, S. B.; Fisher, N. S.; Maser, J.; Vogt, S.; Jacobsen, C.; Tovar-Sanchez, A.; Sanudo-Wihelmy, S. A.; Experimental Facilities Division; Stony Brook Univ.

    2003-01-01

    The study of trace metal cycling by aquatic protists is limited by current analytical techniques. Standard 'bulk' element analysis techniques that rely on physical separations to concentrate cells for analysis cannot separate cells from co-occurring detrital material or other cells of differing taxonomy or trophic function. Here we demonstrate the ability of a synchrotron-based X-ray fluorescence (SXRF) microprobe to quantify the elements Si, Mn, Fe, Ni, and Zn in individual aquatic protist cells. This technique distinguishes between different types of cells in an assemblage and between cells and other particulate matter. Under typical operating conditions, the minimum detection limits are 7.0 x 10{sup -16} mol {mu}m{sup -2} for Si and between 5.0 x 10{sup -20} and 3.9 x 10{sup -19} mol {mu}m{sup -2} for Mn, Fe, Ni, and Zn; this sensitivity is sufficient to detect these elements in cells from even the most pristine waters as demonstrated in phytoplankton cells collected from remote areas of the Southern Ocean. Replicate analyses of single cells produced variations of <5% for Si, Mn, Fe, and Zn and <10% for Ni. Comparative analyses of cultured phytoplankton cells generally show no significant differences in cellular metal concentrations measured with SXRF and standard bulk techniques (spectrophotometry and graphite furnace atomic absorption spectrometry). SXRF also produces two-dimensional maps of element distributions in cells, thereby providing information not available with other analytical approaches. This technique enables the accurate and precise measurement of trace metals in individual aquatic protists collected from natural environments.

  7. Validation of a Geant4 model of the X-ray fluorescence microprobe at the Australian Synchrotron.

    PubMed

    Dimmock, Matthew Richard; de Jonge, Martin Daly; Howard, Daryl Lloyd; James, Simon Alexander; Kirkham, Robin; Paganin, David Maurice; Paterson, David John; Ruben, Gary; Ryan, Chris Gregory; Brown, Jeremy Michael Cooney

    2015-03-01

    A Geant4 Monte Carlo simulation of the X-ray fluorescence microprobe (XFM) end-station at the Australian Synchrotron has been developed. The simulation is required for optimization of the scan configuration and reconstruction algorithms. As part of the simulation process, a Gaussian beam model was developed. Experimental validation of this simulation has tested the efficacy for use of the low-energy physics models in Geant4 for this synchrotron-based technique. The observed spectral distributions calculated in the 384 pixel Maia detector, positioned in the standard back-scatter configuration, were compared with those obtained from experiments performed at three incident X-ray beam energies: 18.5, 11.0 and 6.8 keV. The reduced χ-squared (\\chi^{2}_{\\rm{red}}) was calculated for the scatter and fluorescence regions of the spectra and demonstrates that the simulations successfully reproduce the scatter distributions. Discrepancies were shown to occur in the multiple-scatter tail of the Compton continuum. The model was shown to be particularly sensitive to the impurities present in the beryllium window of the Maia detector and their concentrations were optimized to improve the \\chi^{2}_{\\rm{red}} parametrization in the low-energy fluorescence regions of the spectra.

  8. Laser-excited fluorescence of rare earth elements in fluorite: Initial observations with a laser Raman microprobe

    USGS Publications Warehouse

    Burruss, R.C.; Ging, T.G.; Eppinger, R.G.; Samson, a.M.

    1992-01-01

    Fluorescence emission spectra of three samples of fluorite containing 226-867 ppm total rare earth elements (REE) were excited by visible and ultraviolet wavelength lines of an argon ion laser and recorded with a Raman microprobe spectrometer system. Narrow emission lines ( 0.9 for Eu2+ and 0.99 for Er3+. Detection limits for three micrometer spots are about 0.01 ppm Eu2+ and 0.07 ppm Er3+. These limits are less than chondrite abundance for Eu and Er, demonstrating the potential microprobe analytical applications of laser-excited fluorescence of REE in fluorite. However, application of this technique to common rock-forming minerals may be hampered by competition between fluorescence emission and radiationless energy transfer processes involving lattice phonons. ?? 1992.

  9. The BioCAT Microprobe for X-Ray Fluorescence Imaging, MicroXAFS and Microdiffraction Studies on Biological Samples

    SciTech Connect

    Barrea, R.A.; Gore, D.; Kondrashkina, E.; Weng, T.; Heurich, R.; Vukonich, M.; Orgel, J.; Davidson, M.; Collingwood, J.F.; Mikhaylova, A.; Irving, T.C.

    2007-07-31

    Microbeam capabilities have been recently added to the Biophysics Collaborative Access Team (BioCAT) beamline 18-ID at the Advanced Photon Source to allow x-ray elemental mapping, micro x-ray absorption fine structure and microdiffraction studies on biological samples. The microprobe setup comprises a pair of platinum coated silicon KB mirrors; a sample holder mounted in a high precision positioner (100 nm accuracy); fluorescence detectors including a Si drift detector, Fe and Zn Bent Laue analyzers and a Ge detector; and a CCD detector for micro-diffraction experiments. The energy range of the microprobe is from 3.5 keV up to 17 keV. The fast scanning capabilities of the Bio-CAT beamline facilitate rapid acquisition of x-ray elemental images and micro-XAFS spectra. This paper reports the results of commissioning the KB mirror system and its performance in initial x-ray fluorescence mapping and micro-diffraction studies.

  10. Asymmetric Distribution of Metals in the Xenopus Laevis Oocyte: a Synchrotron X-Ray Fluorescence Microprobe Study

    SciTech Connect

    Popescu, B.F.Gh.; Belak, Z.R.; Ignatyev, K.; Ovsenek, N.; Nichol, H.

    2009-06-04

    The asymmetric distribution of many components of the Xenopus oocyte, including RNA, proteins, and pigment, provides a framework for cellular specialization during development. During maturation, Xenopus oocytes also acquire metals needed for development, but apart from zinc, little is known about their distribution. Synchrotron X-ray fluorescence microprobe was used to map iron, copper, and zinc and the metalloid selenium in a whole oocyte. Iron, zinc, and copper were asymmetrically distributed in the cytoplasm, while selenium and copper were more abundant in the nucleus. A zone of high copper and zinc was seen in the animal pole cytoplasm. Iron was also concentrated in the animal pole but did not colocalize with zinc, copper, or pigment accumulations. This asymmetry of metal deposition may be important for normal development. Synchrotron X-ray fluorescence microprobe will be a useful tool to examine how metals accumulate and redistribute during fertilization and embryonic development.

  11. Asymmetri Distribution of Metals in the Xenopus Laevis Oocyte: a Synchrotron X-Ray Fluorescence Microprobe Study

    SciTech Connect

    Popescu, B.F.G.; Belak, Z.R.; Ignatyev, K.; Ovsenek, N.; Nichol, H.; /Saskatchewan U. /SLAC, SSRL

    2009-04-29

    The asymmetric distribution of many components of the Xenopus oocyte, including RNA, proteins, and pigment, provides a framework for cellular specialization during development. During maturation, Xenopus oocytes also acquire metals needed for development, but apart from zinc, little is known about their distribution. Synchrotron X-ray fluorescence microprobe was used to map iron, copper, and zinc and the metalloid selenium in a whole oocyte. Iron, zinc, and copper were asymmetrically distributed in the cytoplasm, while selenium and copper were more abundant in the nucleus. A zone of high copper and zinc was seen in the animal pole cytoplasm. Iron was also concentrated in the animal pole but did not colocalize with zinc, copper, or pigment accumulations. This asymmetry of metal deposition may be important for normal development. Synchrotron X-ray fluorescence microprobe will be a useful tool to examine how metals accumulate and redistribute during fertilization and embryonic development.

  12. Quantification and localization of trace metals in natural plancton using a synchrotron x-ray fluorescence microprobe.

    SciTech Connect

    Twining, B. S.; Baines, S. B.; Fisher, N. S.; Jacobsen, C.; Maser, J.; State Univ. of New York at Stony Brook

    2003-03-01

    The accumulation of trace metals by planktonic protists influences the growth of primary producers, metal biogeochemical cycling, and metal bioaccumulation in aquatic food chains. Despite their importance, unequivocal measurements of trace element concentrations in individual plankton cells have not been possible to date. We have used the 2-ID-E side-branch hard x-ray microprobe at the Advanced Photon Source to measure trace elements in individual marine plankton cells. This microprobe employs zoneplate optics to produce the sub-micron spatial resolution and low background fluorescence required to produce trace element maps of planktonic protist cells ranging in size from 3 to >50 {micro}m. We have developed preservation, rinsing, and mounting protocols that remove most of the salt from our marine samples, thus simplifying the identification of unknown cells and reducing high Cl-related background fluorescence. We have also developed spectral modeling techniques that account for the frequent overlap of adjacent fluorescence peaks and non-uniform detector response. Finally, we have used parallel soft x-ray transmission and epifluorescence microscopy images to estimate C normalized trace element concentrations, identify functional cell types (e.g., photosynthetic vs. non-photosynthetic), and correlate cell structures with spatial patterns in trace element fluorescence.

  13. Mapping Metal Elements of Shuangbai Dinosaur Fossil by Synchrotron X-ray Fluorescence Microprobe

    SciTech Connect

    Wang, Y.; Qun, Y; Ablett, J

    2008-01-01

    The metal elements mapping of Shuangbai dinosaur fossil, was obtained by synchrotron x-ray fluorescence (SXRF). Eight elements, Ca, Mn, Fe, Cu, Zn, As, Y and Sr were determined. Elements As and Y were detected for the first time in the dinosaur fossil. The data indicated that metal elements are asymmetrical on fossil section. This is different from common minerals. Mapping metals showed that metal element As is few. The dinosaur most likely belongs to natural death. This is different from Zigong dinosaurs which were found dead from poisoning. This method has been used to find that metals Fe and Mn are accrete, and the same is true for Sr and Y. This study indicated that colloid granule Fe and Mn, as well as Sr and Y had opposite electric charges in lithification process of fossils. By this analysis, compound forms can be ascertained. Synchrotron light source x-ray fluorescence is a complementary method that shows mapping of metal elements at the dinosaur fossil, and is rapid, exact and intuitionist. This study shows that dinosaur fossil mineral imaging has a potential in reconstructing the paleoenvironment and ancient geology.

  14. [Mapping metal elements of Shuangbai dinosaur fossil by synchrotron X-ray fluorescence microprobe].

    PubMed

    Wang, Yi-Lin; Yang, Qun; Ablett, J M

    2008-05-01

    The metal elements mapping of Shuangbai dinosaur fossil, was obtained by synchrotron x-ray fluorescence (SXRF). Eight elements, Ca, Mn, Fe, Cu, Zn, As, Y and Sr were determined. Elements As and Y were detected for the first time in the dinosaur fossil. The data indicated that metal elements are asymmetrical on fossil section. This is different from common minerals. Mapping metals showed that metal element As is few. The dinosaur most likely belongs to natural death. This is different from Zigong dinosaurs which were found dead from poisoning. This method has been used to find that metals Fe and Mn are accrete, and the same is ture for Sr and Y. This study indicated that colloid granule Fe and Mn, as well as Sr and Y had opposite electric charges in lithification process of fossils. By this analysis, compound forms can be ascertained. Synchrotron light source x-ray fluorescence is a complementary method that shows mapping of metal elements at the dinosaur fossil, and is rapid, exact and intuitionist. This study shows that dinosaur fossil mineral imaging has a potential in reconstructing the paleoenvironment and ancient geology.

  15. Development of a High Resolution-High Sensitivity Ion Microprobe Facility for Cosmochemical Applications

    NASA Technical Reports Server (NTRS)

    McKeegan, Kevin D.

    1998-01-01

    NASA NAGW-4112 has supported development of the CAMECA ims 1270 ion microprobe at UCLA for applications in cosmochemistry. The instrument has been brought to an operational status and techniques developed for accurate, precise microbeam analysis of oxygen isotope ratios in polished thin-sections. We made the first oxygen isotopic (delta(18)O and delta(17)O) measurements of rare mafic silicates in the most chemically primitive meteorites, the a chondrites (Leshin et al., 1997). The results have implications for both high temperature processing in the nebula and low-T aqueous alteration on the CI asteroid. We have performed measurements of oxygen isotopic compositions of magnetite and co-existing olivine from carbonaceous (Choi et al., 1997) and unequilibrated ordinary chondrites (Choi et al., in press). This work has identified a significant new oxygen isotope reservoir in the early solar system: water characterized by a very high Delta(17)) value of approx. 5 % per thousand. We have determined the spatial distributions of oxygen isotopic anomalies in all major mineral phases of a type B CAI from Allende. We have also studied an unusual fractionated CAI from Leoville and made the first oxygen isotopic measurements in rare CAIs from ordinary chondrites.

  16. Subgroup report on hard x-ray microprobes

    SciTech Connect

    Ice, G.E.; Barbee, T.; Bionta, R.; Howells, M.; Thompson, A.C.; Yun, W.

    1994-09-01

    The increasing availability of synchrotron x-ray sources has stimulated the development of advanced hard x-ray (E{>=}5 keV) microprobes. New x-ray optics have been demonstrated which show promise for achieving intense submicron hard x-ray probes. These probes will be used for extraordinary elemental detection by x-ray fluorescence/absorption and for microdiffraction to identify phase and strain. The inherent elemental and crystallographic sensitivity of an x-ray microprobe and its inherently nondestructive and penetrating nature makes the development of an advanced hard x-ray microprobe an important national goal. In this workshop state-of-the-art hard x-ray microprobe optics were described and future directions were discussed. Gene Ice, Oak Ridge National Laboratory (ORNL), presented an overview of the current status of hard x-ray microprobe optics and described the use of crystal spectrometers to improve minimum detectable limits in fluorescent microprobe experiments. Al Thompson, Lawrence Berkeley Laboratory (LBL), described work at the Center for X-ray Optics to develop a hard x-ray microprobe based on Kirkpatrick-Baez (KB) optics. Al Thompson also showed the results of some experimental measurements with their KB optics. Malcolm Howells presented a method for bending elliptical mirrors and Troy Barbee commented on the use of graded d spacings to achieve highest efficiency in KB multilayer microfocusing. Richard Bionta, Lawrence Livermore National Laboratory (LLNL), described the development of the first hard x-ray zone plates and future promise of so called {open_quotes}jelly roll{close_quotes} or sputter slice zone plates. Wenbing Yun, Argonne National Laboratory (ANL), described characterization of jelly roll and lithographically produced zone plates and described the application of zone plates to focus extremely narrow bandwidths by nuclear resonance. This report summarizes the presentations of the workshop subgroup on hard x-ray microprobes.

  17. The proton (nuclear) microprobe

    NASA Astrophysics Data System (ADS)

    Legge, G. J. F.

    1989-04-01

    The scanning proton microprobe (SPMP) is closely related to the scanning electron microprobe (SEMP) or scanning electron microscope (SEM) with X-ray detector. Though the much greater elemental sensitivity of the SPMP is inherent in the physics, the generally inferior spatial resolution of the SPMP is not inherent and big improvements are possible, As its alternative name would imply, the SPMP is often used with heavier particle beams and with nuclear rather than atomic reactions. Its versatility and quantitative accuracy have justified greater instrumentation and computer power than that associated with other microprobes. It is fast becoming an industrially and commercially important instrument and there are few fields of scientific research in which it has not played a part. Notable contributions have been made in biology, medicine, agriculture, semiconductors, geology, mineralogy, extractive metallurgy, new materials, archaeology, forensic science, catalysis, industrial problems and reactor technology.

  18. Isotopic Investigations of Nebular and Parent Body Processes with a High Sensitivity Ion Microprobe

    NASA Technical Reports Server (NTRS)

    McKeegan, Kevin D.

    2005-01-01

    NASA supported the development of the CAMECA ims 1270 ion microprobe at UCLA for applications in cosmochemistry. The primary investigations centered on measuring the microscopic distributions of key isotopic abundances in primitive meteoritic materials as a means of constraining the nature of important thermal and chemical processes in the solar nebula and the timescales associated with those processes. Our prior work on oxygen isotope anomalies in a wide variety of meteoritic materials had led us to a view of a spatially heterogeneous nebula, and in particular, a restricted region for CAI formation that is characterized by O-16-rich gas. Because of its production of CAIs in the energetic local environment near the protosun, the existence of a natural transport mechanism via bipolar outflows, and a general astrophysical plausibility, we were attracted to the fluctuating X-wind model which had been put forward by Frank Shu, Typhoon Lee, and colleagues. With our collaborators, we undertook a series of investigations to test the viability of this hypothesis; this work led directly to the discovery of live Be in CAIs and a clear demonstration of the existence of 160-rich condensates, which necessarily implies an O-16-rich gaseous reservoir in the nebula. Both of these observations fit well within the context of X-wind type models, i.e. formation of CAIs (or condensation of their precursors) in the reconnection ring sunward of the inner edge of the accretion disk, however much work remains to be done to test whether the physical parameters of the model can quantitatively predict not only the thermal histories of CAIs but also their radioactivity. The issue of spatial heterogeneity in the nebula, central to the X-wind model, is also at the heart of any chronology based on short-lived radioisotopes. In this work, we followed up on strong hints for presence of exireme:j: (53 day) short-lived Be-7, and have prepared a manuscript (in revision). We also measured A1-Mg

  19. Development of an x-ray fluorescence microprobe at the National Synchrotron Light Source, Brookhaven National Laboratory: Early results: Comparison with data from other techniques

    SciTech Connect

    Smith, J.V.; Rivers, M.L.; Sutton, S.R.; Jones, K.W.; Hanson, A.L.; Gordon, B.M.

    1986-01-01

    Theoretical predictions for the detection levels in x-ray fluorescence analysis with a synchrotron storage ring are being achieved experimentally at several laboratories. This paper is deliberately restricted to the state of development of the Brookhaven National Laboratory/University of Chicago instruments. Analyses at the parts per million (ppM) level are being made using white light apertured to 20 ..mu..m and an energy dispersive system. This system is particularly useful for elements with Z > 20 in materials dominated by elements with Z < 20. Diffraction causes an interference for crystalline materials. Development of a focusing microprobe for tunable monochromatic x-rays and a wavelength dispersive spectrometer (WDS) is delayed by problems in shaping an 8:1 focusing mirror to the required accuracy. Reconnaissance analyses with a wiggler source on the CHESS synchrotron have been made in the K spectrum up to Z = 80.

  20. Position-sensitive scanning fluorescence correlation spectroscopy.

    PubMed

    Skinner, Joseph P; Chen, Yan; Müller, Joachim D

    2005-08-01

    Fluorescence correlation spectroscopy (FCS) uses a stationary laser beam to illuminate a small sample volume and analyze the temporal behavior of the fluorescence fluctuations within the stationary observation volume. In contrast, scanning FCS (SFCS) collects the fluorescence signal from a moving observation volume by scanning the laser beam. The fluctuations now contain both temporal and spatial information about the sample. To access the spatial information we synchronize scanning and data acquisition. Synchronization allows us to evaluate correlations for every position along the scanned trajectory. We use a circular scan trajectory in this study. Because the scan radius is constant, the phase angle is sufficient to characterize the position of the beam. We introduce position-sensitive SFCS (PSFCS), where correlations are calculated as a function of lag time and phase. We present the theory of PSFCS and derive expressions for diffusion, diffusion in the presence of flow, and for immobilization. To test PSFCS we compare experimental data with theory. We determine the direction and speed of a flowing dye solution and the position of an immobilized particle. To demonstrate the feasibility of the technique for applications in living cells we present data of enhanced green fluorescent protein measured in the nucleus of COS cells.

  1. U-Pb geochronology of zircons form lunar Breccia 73217 using a sensitive high mass-resolution ion microprobe

    NASA Astrophysics Data System (ADS)

    Compston, W.; Williams, I. S.; Meyer, C.

    1984-02-01

    U-Pb age determinations on four lunar zircons from existing thin-sections of one highland breccia, 73217, using the recently constructed ion microprobe SHRIMP, are reported. The analytical reproducibility of SHRIMP is demonstrated, and procedures for measuring Pb/U, Th/U, and corecting for initial Pb are explained. Electron microprobe analyses for the zircons are also are reported. The results show that the four zircons survived the lunar cataclysm without any identifiable effects on their U-Pb systematics. All four indicate a single age of 4356 +23 or -14 m.y. The zircons have experienced small variable amounts of Pb loss since crystallization, from almost zero up to about 10 percent. If this occurred during one later event, then age of the latter is between 1100 and 2300 m.y.

  2. U-Pb geochronology of zircons form lunar Breccia 73217 using a sensitive high mass-resolution ion microprobe

    NASA Technical Reports Server (NTRS)

    Compston, W.; Williams, I. S.; Meyer, C.

    1984-01-01

    U-Pb age determinations on four lunar zircons from existing thin-sections of one highland breccia, 73217, using the recently constructed ion microprobe SHRIMP, are reported. The analytical reproducibility of SHRIMP is demonstrated, and procedures for measuring Pb/U, Th/U, and corecting for initial Pb are explained. Electron microprobe analyses for the zircons are alsoar reported. The results show that the four zircons survived the lunar cataclysm without any identifiable effects on their U-Pb systematics. All four indicate a single age of 4356 +23 or -14 m.y. The zircons have experienced small variable amounts of Pb loss since crystallization, from almost zero up to about 10 percent. If this occurred during one later event, then age of the latter is between 1100 and 2300 m.y.

  3. U-Pb geochronology of zircons form lunar Breccia 73217 using a sensitive high mass-resolution ion microprobe

    SciTech Connect

    Compston, W.; Williams, I.S.

    1984-02-15

    U-Pb age determinations on four lunar zircons from existing thin-sections of one highland breccia, 73217, using the recently constructed ion microprobe SHRIMP, are reported. The analytical reproducibility of SHRIMP is demonstrated, and procedures for measuring Pb/U, Th/U, and corecting for initial Pb are explained. Electron microprobe analyses for the zircons are also reported. The results show that the four zircons survived the lunar cataclysm without any identifiable effects on their U-Pb systematics. All four indicate a single age of 4356 +23 or -14 m.y. The zircons have experienced small variable amounts of Pb loss since crystallization, from almost zero up to about 10 percent. If this occurred during one later event, then age of the latter is between 1100 and 2300 m.y. 18 references.

  4. Improved resolution and sensitivity on the ANSTO microprobe and it’s application to μ-PIXE

    NASA Astrophysics Data System (ADS)

    Siegele, R.; Kachenko, A. G.; Ionescu, M.; Cohen, D. D.

    2009-06-01

    We report on the improved spatial resolution of the ANSTO heavy ion microprobe, achieved through the use of a higher brightness ion source for hydrogen. The improved resolution will be demonstrated for applications of μ-PIXE. With the higher brightness source, a 3 μm resolution was achieved for μ-PIXE elemental analysis. This is illustrated in high resolution images of nickel (Ni)-hyperaccumulating Hybanthus floribundus subsp. floribundus leaf tissues, where individual cells were clearly visible in the acquired elemental images. The higher resolution images illustrated that Ni was localised in epidermal cell walls.

  5. Fluorescence microscopy studies on ALA-sensitized tissues

    NASA Astrophysics Data System (ADS)

    Huettmann, Gereon; Achtelik, Wolfgang; Loening, Martin; Sommer, Konrad; Diddens, Heyke C.

    1996-12-01

    Fluorescence microscopy has the potential to study the spatial distribution of photosensitizers in tissue samples with cellular or subcellular resolution. A fluorescence microscope was developed to study the distribution of photosensitizer in tissue samples by acquiring fluorescence images in various spectral ranges and spatially resolved fluorescence spectra both from identical samples. Both methods provide complementary information, since the fluorescence images show the distribution of the sensitizers with a high spatial resolution whereas spatially resolved fluorescence spectra can identify the sensitizers and separate their fluorescence from background light emission by the spectral shape of the fluorescence. Protoporphyrin IX (PPIX) distribution induced by 5-aminolevulinic acid (ALA) was studied by fluorescence microscopy in basal cell carcinoma (BCC) and in cervical intraepithelial neoplasia (CIN). In an attempt to understand the varying success in treating BCC with topically applied ALA the PPIX distribution was studied in BCC samples of 10 patients. A strong fluorescence was observed in tumor cells as well as in epidermis, sebaceous glands, and hair follicles. The depth of PPIX sensitization of the BCCs ranged from 0.4 to 3 mm and the ratio of tumor versus epidermal fluorescence of uninvolved skin was near one. In the BCCs an uneven sensitization with a lower fluorescence in the center of the tumor was often observed. Samples of the cervical mucosa also showed PPIX fluorescence in the endothelial layer, the malignant tissues and the glands. No increased fluorescence of the dysplastic lesions compared to the epithelium was observed.

  6. Spectral light measurements in microbenthic phototrophic communities with a fiber-optic microprobe coupled to a sensitive diode array detector

    SciTech Connect

    Kuehl, M. ); Joergensen, B.B. )

    1992-12-01

    A diode array detector system for microscale light measurements with fiber-optic microprobes was developed; it measures intensities of 400-900-nm light over >6 orders of magnitude with a spectral resolution of 2-5 nm. Fiber-optic microprobes to measure field radiance or scalar irradiance were coupled to the detector system and used for spectral light measurements in hypersaline microbial mats and in laminated phototrophic communities of coastal sediments. The vertical distribution of major photopigments of microalgae, cyanobacteria, and anoxygenic phototrophic bacteria could be identified from extinction maxima in measured radiance spectra at 430-550 nm (Chl a and carotenoids), 620-625 nm (phycocyanin), 675 nm (Chl a), 745-750 nm (BChl c), 800-810 nm, and 860-880 nm (BChl a). Scalar irradiance spectra exhibited a different spectral composition and a higher light intensity at the sediment surface as compared to incident light. IR light thus reached 200% of incident at the sediment surface. Maximal light penetration was found for IR light, whereas visible light was strongly attenuated in the upper 0-2 mm of the sediment. Measurements of photon scalar irradiance (400-700 nm) were combined with microelectrode measurements of oxygenic photosynthesis in the coastal sediment. With an incident light intensity of 200 [mu]Einst m[sup [minus]2]s[sup [minus]1], photon scalar irradiance reached a maximum of 283 [mu]Einst m[sup [minus]2]s[sup [minus]1] at the sediment surface. The lower boundary of the euphotic zone was 2.2 mm below the surface at a light intensity of 12 [mu]Einst m[sup [minus]2]s[sup [minus]1]. 20 refs., 6 figs.

  7. Materials analysis with a nuclear microprobe

    SciTech Connect

    Maggiore, C.J.

    1980-01-01

    The ability to produce focused beams of a few MeV light ions from Van de Graaff accelerators has resulted in the development of nuclear microprobes. Rutherford backscattering, nuclear reactions, and particle-induced x-ray emission are used to provide spatially resolved information from the near surface region of materials. Rutherford backscattering provides nondestructive depth and mass resolution. Nuclear reactions are sensitive to light elements (Z < 15). Particle-induced x-ray analysis is similar to electron microprobe analysis, but 2 orders of magnitude more sensitive. The focused beams are usually produced with specially designed multiplets of magnetic quadrupoles. The LASL microprobe uses a superconducting solenoid as a final lens. The data are acquired by a computer interfaced to the experiment with CAMAC. The characteristics of the information acquired with a nuclear microprobe are discussed; the means of producing the beams of nuclear particles are described; and the limitations and applications of such systems are given.

  8. Development of a Laboratory Micron-Resolution X-ray Microprobe to Map Mineralogy and Trace Elements at PPM Sensitivity for Digital Rock, Magma, and Mining Applications

    NASA Astrophysics Data System (ADS)

    Yun, W.; Lewis, S.; Stripe, B.; Chen, S.; Reynolds, D.; Spink, I.; Lyon, A.

    2015-12-01

    We are developing a patent-pending x-ray microprobe with substantially unprecedented performance attributes: <5 μm spot on the sample (with 1 μm targeted), large working distances of >2 cm, narrow spectral bandwidth, and large x-ray flux. The outstanding performance is enabled by: (1) a revolutionary new type of high flux x-ray source designed to be >10X brighter than the brightest rotating anode x-ray source available; (2) an axially symmetric x-ray mirror lens with large solid angle collection and high focusing efficiency; and (3) a detector configuration that enables the collection of 10X more x-rays than current microXRF designs. The sensitivity will be ppm-scale, far surpassing charged particle analysis (e.g. EPMA and SEM-EDS), and >1000X throughput over the leading micro-XRFs. Despite the introduction of a number of laboratory microXRF systems in the past decade, the state-of-the-art has been limited primarily by low resolution (~30 μm) and low throughput. This is substantially attributable to a combination of low x-ray source brightness and poor performance x-ray optics. Here we present our initial results in removing the x-ray source bottleneck, in which we use a novel x-ray source using Fine Anode Array Source Technology (Sigray FAAST™). When coupled with our proprietary high efficiency x-ray mirror lens, the throughput achieved is comparable to that of many synchrotron microXRF beamlines. Potential applications of the x-ray microprobe include high throughput mapping of mineralogy at high resolution, including trace elements, such as rare earth metals, and deposits (e.g. siderite, clays), with ppm sensitivity, providing information for properties such as permeability and elastic/mechanical properties, and to provide compositional information for Digital Rock. Additional applications include those in which the limited penetration of electrons limits achieving adequate statistics, such as determining the concentration of precious minerals in mine

  9. An X-ray microprobe facility using synchrotron radiation.

    PubMed

    Gordon, B M; Jones, K W; Hanson, A L; Pounds, J G; Rivers, M L; Spanne, P; Sutton, S R

    1990-01-01

    An X-ray microprobe for trace elemental analysis at micrometer spatial resolutions, using synchrotron radiation (SR), is under development. The facility consists of two beamlines, one including a 1:1 focusing mirror and the other an 8:1 ellipsoidal mirror. At present, "white light" is used for excitation of the characteristic X-ray fluorescence lines. Sensitivities in thin biological samples are in the range of 2-20 fg in 100 microns2 areas in 5 min irradiation times. Scanning techniques, as well as microtomography and chemical speciation, are discussed. Application to a specific biomedical study is included.

  10. An x-ray microprobe facility using synchrotron radiation

    SciTech Connect

    Gordon, B.M.; Jones, K.W.; Hanson, A.L.; Pounds, J.G.; Rivers, M.L.; Spanne, P.; Sutton, S.R.

    1989-01-01

    A x-ray microprobe for trace elemental analysis at micrometer spatial resolutions using synchrotron radiation (SR) is under development. The facility consists of two beamlines, one including a 1:1 focusing mirror and the other an 8:1 ellipsoidal mirror. At present ''white light''' is used for excitation of the characteristic x-ray fluorescence lines. Sensitivities in thin biological samples are in the range of 2-20 fg in 100 ..mu..m/sup 2/ areas in 5 min irradiation times. Scanning techniques as well as microtomography and chemical speciation are discussed. Application to a specific biomedical study is included. 13 refs., 2 figs.

  11. Position sensitive detector for fluorescence lifetime imaging

    NASA Astrophysics Data System (ADS)

    Prokazov, Y.; Turbin, E.; Weber, A.; Hartig, R.; Zuschratter, W.

    2014-12-01

    We present a detector system with a microchannel plate based photomultiplier tube (MCP-PMT) and its application for fluorescence lifetime imaging (FLIM) in visible light. A capacity coupled imaging technique (charge image) combined with a charge division anode is employed for the positional readout. Using an artificial neural network's (ANN) computation model we are able to reconstruct the position of the incident photon as precise as 20 microns over the detector active area of 25 mm diameter. Thus, the resulting image quality corresponds roughly to a megapixel conventional CCD camera. Importantly, it is feasible to reach such resolution using only 9 charge acquisition channels supporting the anode structure of 14 interconnected readout electrodes. Additionally, the system features better than 50 ps temporal resolution allowing single photon counting FLIM acquisition with a regular fluorescence wide-field microscope.

  12. Octyl gallate: An antioxidant demonstrating selective and sensitive fluorescent property.

    PubMed

    Wang, Qing; Zhang, Yongkui; Li, Hui

    2017-03-15

    Octyl gallate (OG) is an internationally recognized antioxidant that demonstrates selective and sensitive fluorescent property. The fluorescence of OG can be selectively enhanced in the presence of human serum albumin (HSA) and bovine serum albumin (BSA). The specific structures of HSA and BSA provided the basic conditions for fluorescence enhancement. OG yielded approximately 49- and 11-fold increments in emission intensity in the presence of HSA and BSA at a molar ratio of 1:1, respectively. The lifetimes of HSA and BSA correspondingly decreased. A Förster resonance energy transfer phenomenon occurred during interaction between OG and HSA or BSA. Our in-depth investigation of OG-HSA interaction showed that formation of a stable complex was an important prerequisite to efficiently enhance the fluorescence of OG. The selective and sensitive fluorescent property of OG can possibly be used to determine OG concentration via the standard addition method, which must be performed under certain conditions.

  13. Sensitive turn-on fluorescent detection of tartrazine based on fluorescence resonance energy transfer.

    PubMed

    Huang, Sheng Tian; Shi, Yan; Li, Nian Bing; Luo, Hong Qun

    2012-01-18

    We introduce a sensitive, rapid, label-free and general fluorescent method for the determination of tartrazine by competitive binding to reduced graphene oxide (rGO) against fluorescein, and the fluorescence recovery upon fluorescein desorption from rGO provides a quantitative readout for tartrazine, giving a detection limit of 0.53 ng mL(-1).

  14. Ultra-Sensitive Nanofiber Fluorescence Detection in a Microfluidic Chip

    PubMed Central

    Li, Zhiyong; Xu, Yingxin; Fang, Wei; Tong, Limin; Zhang, Lei

    2015-01-01

    We report an ultra-sensitive and robust fluorescence sensor made by using a biconical taper with a waist diameter of 720 nm for both excitation and fluorescence collection. To enhance the stability of the fluorescence sensor, the biconical taper has been embedded in a 125 µm wide microchannel with a detection length of 2.5 cm. Investigated by measuring the fluorescence intensity of rhodamine 6G (R6G), the sensor shows a detection limit down to 100 pM, with excellent reversibility in a concentration range of 0–10 nM. The sensor has also been applied to quantum dot (QD)-labeled streptavidin measurements, yielding a detection sensitivity down to 10 pM for QDs. In addition, the small sample volume (ca. 500 nL), high sampling throughput, and seamless connection between the biconical taper and standard optical fibers offer a number of attractive advantages for chemical and biosensing applications. PMID:25808762

  15. Coolwater culmination: Sensitive high-resolution ion microprobe (SHRIMP) U-Pb and isotopic evidence for continental delamination in the Syringa Embayment, Salmon River suture, Idaho

    USGS Publications Warehouse

    Lund, K.; Aleinikoff, J.N.; Yacob, E.Y.; Unruh, D.M.; Fanning, C.M.

    2008-01-01

    During dextral oblique translation along Laurentia in western Idaho, the Blue Mountains superterrane underwent clockwise rotation and impinged into the Syringa embayment at the northern end of the Salmon River suture. Along the suture, the superterrane is juxtaposed directly against western Laurentia, making this central Cordilleran accretionary-margin segment unusually attenuated. In the embayment, limited orthogonal contraction produced a crustal wedge of oceanic rocks that delaminated Laurentian crust. The wedge is exposed through Laurentian crust in the Coolwater culmination as documented by mapping and by sensitive high-resolution ion microprobe U-Pb, Sri, and ??Nd data for gneisses that lie inboard of the suture. The predominant country rock is Mesoproterozoic paragneiss overlying Laurentian basement. An overlying Neoproterozoic (or younger) paragneiss belt in the Syringa embayment establishes the form of the Cordilleran miogeocline and that the embayment is a relict of Rodinia rifting. An underlying Cretaceous paragneiss was derived from arc terranes and suture-zone orogenic welt but also from Laurentia. The Cretaceous paragneiss and an 86-Ma orthogneiss that intruded it formed the wedge of oceanic rocks that were inserted into the Laurentian margin between 98 and 73 Ma, splitting supracrustal Laurentian rocks from their basement. Crustal thickening, melting and intrusion within the wedge, and folding to form the Coolwater culmination continued until 61 Ma. The embayment formed a restraining bend at the end of the dextral transpressional suture. Clockwise rotation of the impinging superterrane and overthrusting of Laurentia that produced the crustal wedge in the Coolwater culmination are predicted by oblique collision into the Syringa embayment. Copyright 2008 by the American Geophysical Union.

  16. Determination of ethambutol by a sensitive fluorescent probe

    NASA Astrophysics Data System (ADS)

    Wu, Wen-Ying; Yang, Ji-Yuan; Du, Li-Ming; Wu, Hao; Li, Chang-Feng

    2011-08-01

    The competitive reaction between ethambutol and two fluorescent probes (i.e., berberine and palmatine) for occupancy of the cucurbit[7]uril (CB[7]) cavity was studied by spectrofluorometry. The CB[7] reacts with these probes to form stable complexes, and the fluorescence intensity of the complexes is greatly enhanced. In addition, the excitation and emission wavelengths of their complexes moved to wavelengths of 343 nm and 495 nm, respectively. However, the addition of ethambutol dramatically quenches the fluorescence intensity of the two complexes. Accordingly, a couple of new fluorescence quenching methods for the determination of ethambutol were established. The methods can be applied for quantifying ethambutol. A linear relationship between the fluorescence quenching values (Δ F) and ethambutol concentration exists in the range of 5.0-1000.0 ng mL -1, with a correlation coefficient ( r) of 0.9997. The detection limit is 1.7 ng mL -1. The fluorescent probe of berberine has higher sensitivity than palmatine. This paper also discusses the mechanism of fluorescence indicator probes.

  17. Sensitivity of activatable reactive oxygen species probes by fluorescence spectroelectrochemistry

    PubMed Central

    Wang, Steven T.; Zhegalova, Natalia G.; Gustafson, Tiffany P.; Zhou, Andy; Sher, Joel; Achilefu, Samuel; Berezinand, Oleg Y.

    2013-01-01

    We have developed a new analytical method of evaluating activatable fluorescent probes for ROS detection using integrated fluorescence spectroelectrochemistry. Tafel formalism was applied to describe the process of the probes’ oxidation under electrochemical conditions and identify a novel parameter defined as the threshold oxidation potential. This potential can serve as an approximation to the equilibrium potential and can be utilized for determining the sensitivity of a probe to oxidation. Based upon the measured values of threshold potentials, the order of sensitivity towards oxidation among several mostly used probes was determined to be following (from highest to lowest): 2,7-dichlorodihydrofluorescein > dihydroethidium > dihydrorhodamine 123 > dihydrorhodamine 6G. The presented approach opens up a new direction in synthesizing and screening novel ROS probes with a well-defined sensitivity for in vitro and in vivo applications. PMID:23736882

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

    NASA Astrophysics Data System (ADS)

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

    2005-06-01

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

  19. Thermosensitive polymer-modified gold nanoparticles with sensitive fluorescent properties

    NASA Astrophysics Data System (ADS)

    Luo, Chunhua; Dong, Qiujing; Qian, Meijuan; Zhang, Hong

    2016-11-01

    Two types of thermosensitive polymer-modified gold nanoparticles (GNPs), P(DMAM-co-MADMAC)-modified GNPs (GNPs@PDM) and P(NIPAM-co-MADMAC)-modified GNPs (GNPs@PNM), are fabricated by the "grafting through" polymerization technique. The as-prepared GNPs are characterized by UV-vis, TEM, XPS, TGA, FT-IR and 1H NMR spectroscopy. The thermosensitivity and fluorescence of the GNPs are investigated. It is found that all GNPs aqueous solutions but GNPs@PDM1 exhibit thermosensitivity originated from thermosensitive polymer chains and sensitive fluorescence from the dimethylaminochalcone group. The lower critical solution temperature (LCST) of the GNPs decreases with the increasing content of MADMAC unit in the GNPs. The GNPs aqueous solution shows weak fluorescence after the temperature increases from 25 °C to 45 °C, or after β-cyclodextrin (β-CD) is added. Furthermore, it exhibits strong fluorescence when the solvent is changed to ethanol or chloroform, and the fluorescent wavelength undergoes a blue shift from ethanol to chloroform.

  20. Mars Microprobe Entry Analysis

    NASA Technical Reports Server (NTRS)

    Braun, Robert D.; Mitcheltree, Robert A.; Cheatwood, F. McNeil

    1998-01-01

    The Mars Microprobe mission will provide the first opportunity for subsurface measurements, including water detection, near the south pole of Mars. In this paper, performance of the Microprobe aeroshell design is evaluated through development of a six-degree-of-freedom (6-DOF) aerodynamic database and flight dynamics simulation. Numerous mission uncertainties are quantified and a Monte-Carlo analysis is performed to statistically assess mission performance. Results from this 6-DOF Monte-Carlo simulation demonstrate that, in a majority of the cases (approximately 2-sigma), the penetrator impact conditions are within current design tolerances. Several trajectories are identified in which the current set of impact requirements are not satisfied. From these cases, critical design parameters are highlighted and additional system requirements are suggested. In particular, a relatively large angle-of-attack range near peak heating is identified.

  1. Determination of phenformin hydrochloride employing a sensitive fluorescent probe

    NASA Astrophysics Data System (ADS)

    Shi, Lin; Xie, Jian-Hong; Du, Li-Ming; Chang, Yin-xia; Wu, Hao

    2016-06-01

    A complexation of non-fluorescent phenformin hydrochloride (PFH) with cucurbit [7]uril (CB [7]) in aqueous solution was investigated using the fluorescent probe of palmatine (PAL) coupled with CB [7]. The fluorescent probe of CB [7]-PAL exhibited strong fluorescence in aqueous solution, which was quenched gradually with the increase of PFH. This effect is observed because when PFH was added to the host-guest system of CB [7]-PAL, PFH and PAL competed to occupy the CB [7] cavity. Portions of the PAL molecule were expelled from the CB [7] cavity owing to the introduction of PFH. Based on the significant quenching of the supramolecular complex fluorescence intensity, a fluorescence method of high sensitivity and selectivity was developed to determine PFH with good precision and accuracy for the first time. The linear range of the method was 0.005-1.9 μg mL- 1 with a detection limit of 0.003 μg mL- 1. In this work, association constants (K) of PFH with CB [7] were also determined. KCB [7]-PFH = (2.52 ± 0.05) × 105 L mol- 1. The ability of PFH to bind with CB [7] is stronger than that of PAL. The results of a density functional theory calculation authenticated that the moiety of PFH was embedded in the hydrophobic cavity of CB [7] tightly, and the nitrogen atom is located in the vicinity of a carbonyl-laced portal in the energy-minimized structure. The molecular modelling of the interaction between PFH and CB [7] was also confirmed by 1H NMR spectra (Bruker 600 MHz).

  2. Determination of amantadine and rimantadine using a sensitive fluorescent probe

    NASA Astrophysics Data System (ADS)

    Wang, Guang-Quan; Qin, Yan-Fang; Du, Li-Ming; Li, Jun-Fei; Jing, Xu; Chang, Yin-Xia; Wu, Hao

    2012-12-01

    Amantadine hydrochloride (AMA) and rimantadine hydrochloride (RIM) are non-fluorescent in aqueous solutions. This property makes their determination through direct fluorescent method difficult. The competing reactions and the supramolecular interaction mechanisms between the two drugs and coptisine (COP) as they fight for occupancy of the cucurbit[7]uril (CB[7]) cavity, were studied using spectrofluorimetry, 1H NMR, and molecular modeling calculations. Based on the significant quenching of the supramolecular complex fluorescence intensity, a fluorescent probe method of high sensitivity and selectivity was developed to determine AMA or RIM in their pharmaceutical dosage forms and in urine samples with good precision and accuracy. The linear range of the method was from 0.0040 to 1.0 μg mL-1 with a detection limit ranging from 0.0012 to 0.0013 μg mL-1. This shows that the proposed method has promising potential for therapeutic monitoring and pharmacokinetics and for clinical application.

  3. An ultra sensitive fluorescent nanosensor for detection of ionic copper.

    PubMed

    Kacmaz, Sibel; Ertekin, Kadriye; Mercan, Deniz; Oter, Ozlem; Cetinkaya, Engin; Celik, Erdal

    2015-01-25

    A stable and ultra sensitive nano-scale fluorescent chemo-sensor for trace amounts of Cu(2+) was proposed. The Cu(2+) selective fluoroionophore 2-{[(2-aminophenyl)imino]methyl}-4,6-di-tert-butylphenol (DMK-7) was encapsulated in polymeric ethyl cellulose. The sensing membranes were fabricated in form of nanofibers and thin films. When embedded in polymers, the exploited DMK-7 dye exhibited enhanced photophysical characteristics in absorbance, Stoke's shift, fluorescence quantum yield, and short and long-term photostability with respect to the solution phase. Sensing abilities of the nanofibers and thin films were tested by steady state and time resolved fluorescence spectroscopy. To our knowledge, this is the first attempt using the DMK-7-doped electrospun nanofibrous materials for copper sensing. The offered sensor displayed a sensitive response with a detection limit of 3.3×10(-13) M for Cu(2+) ions over a wide concentration range of 5.0×10(-12)-5.0×10(-5). Additionally, exhibited high selectivity over convenient cations; Na(+), K(+), Ca(2+), Mg(2+), NH4(+) and Ag(+), Al(3+), Ba(2+), Co(2+), Cr(3+), Fe(3+), Fe(2+), Hg(2+), Li(+), Mn(2+), Ni(2+), Pb(2+), Sn(2+) and Zn(2+).

  4. An ultra sensitive fluorescent nanosensor for detection of ionic copper

    NASA Astrophysics Data System (ADS)

    Kacmaz, Sibel; Ertekin, Kadriye; Mercan, Deniz; Oter, Ozlem; Cetinkaya, Engin; Celik, Erdal

    2015-01-01

    A stable and ultra sensitive nano-scale fluorescent chemo-sensor for trace amounts of Cu2+ was proposed. The Cu2+ selective fluoroionophore 2-{[(2-aminophenyl)imino]methyl}-4,6-di-tert-butylphenol (DMK-7) was encapsulated in polymeric ethyl cellulose. The sensing membranes were fabricated in form of nanofibers and thin films. When embedded in polymers, the exploited DMK-7 dye exhibited enhanced photophysical characteristics in absorbance, Stoke's shift, fluorescence quantum yield, and short and long-term photostability with respect to the solution phase. Sensing abilities of the nanofibers and thin films were tested by steady state and time resolved fluorescence spectroscopy. To our knowledge, this is the first attempt using the DMK-7-doped electrospun nanofibrous materials for copper sensing. The offered sensor displayed a sensitive response with a detection limit of 3.3 × 10-13 M for Cu2+ ions over a wide concentration range of 5.0 × 10-12-5.0 × 10-5. Additionally, exhibited high selectivity over convenient cations; Na+, K+, Ca2+, Mg2+, NH4+ and Ag+, Al3+, Ba2+, Co2+, Cr3+, Fe3+, Fe2+, Hg2+, Li+, Mn2+, Ni2+, Pb2+, Sn2+ and Zn2+.

  5. Highly sensitive turn-on fluorescence detection of thrombomodulin based on fluorescence resonance energy transfer

    NASA Astrophysics Data System (ADS)

    Kong, Liyan; Zhu, Jiaming; Wang, Wen; Jin, Lehe; Fu, Yanjiao; Duan, Bohui; Tan, Liang

    2017-02-01

    As an integral glycoprotein on the surface of endothelial cells, thrombomodulin (TM) has very high affinity for thrombin. TM has been regarded to be a marker of endothelial damage since it can be released during endothelial cell injury. In this work, a highly sensitive fluorescence method for the quantitative detection of TM was developed. TM antibody (Ab) and bovine serum albumin (BSA) were bound on gold nanoparticles (AuNPs) to construct BSA-AuNPs-Ab nanocomposites and they were characterized by transmission electron microscope and UV-vis spectrophotometry. The fluorescence of acridine orange (AO) was quenched by the prepared gold nanocomposites based on fluorescence resonance energy transfer (FRET). In the presence of TM, the fluorescence was turned on due to the effective separation of AO from the surface of gold nanocomposites. Under optimum conditions, the enhanced fluorescence intensity displayed a linear relationship with the logarithm of the TM concentration from 0.1 pg mL- 1 to 5 ng mL- 1 with a low detection limit of 12 fg mL- 1. The release of soluble thrombomodulin (sTM) by the injured HUVEC-C cells in the presence of H2O2 was investigated using the proposed method. The released sTM content in the growth medium was found to be increased with the enhancement of contact time of the cells with H2O2.

  6. Environment-sensitive behavior of fluorescent molecular rotors

    PubMed Central

    2010-01-01

    Molecular rotors are a group of fluorescent molecules that form twisted intramolecular charge transfer (TICT) states upon photoexcitation. When intramolecular twisting occurs, the molecular rotor returns to the ground state either by emission of a red-shifted emission band or by nonradiative relaxation. The emission properties are strongly solvent-dependent, and the solvent viscosity is the primary determinant of the fluorescent quantum yield from the planar (non-twisted) conformation. This viscosity-sensitive behavior gives rise to applications in, for example, fluid mechanics, polymer chemistry, cell physiology, and the food sciences. However, the relationship between bulk viscosity and the molecular-scale interaction of a molecular rotor with its environment are not fully understood. This review presents the pertinent theories of the rotor-solvent interaction on the molecular level and how this interaction leads to the viscosity-sensitive behavior. Furthermore, current applications of molecular rotors as microviscosity sensors are reviewed, and engineering aspects are presented on how measurement accuracy and precision can be improved. PMID:20843326

  7. Ionic liquid sensitized fluorescence determination of four isoquinoline alkaloids.

    PubMed

    Wu, Hao; Zhang, Li-Bing; Du, Li-Ming

    2011-07-15

    The fluorescence spectra of berberine, palmatine, jatrorrhizine, and coptisine in ionic liquids were studied and found to increase significantly in ionic liquids, with [C(8)MIM][PF(6)] having the greatest increase. Further studies showed that these drugs could be extracted from an aqueous solution by [C(8)MIM][PF(6)] using the temperature-assisted ionic liquid dispersive liquid phase microextraction method. The enrichment factors were 81.8-82.3, and the extraction recovery was 98.5%, 98.1%, 98.3%, and 98.8% for berberine, palmatine, jatrorrhizine, and coptisine, respectively. Based on the [C(8)MIM][PF(6)] preconcentration, separation, and sensitized fluorescence for these drugs, a new selective and sensitive method for the determination of concentration of these four drugs in aqueous samples was presented. At optimum conditions, the linear relationship was obtained in the ranges of 0.8-130 ng mL(-1), 0.9-160 ng mL(-1), 0.7-140 ng mL(-1), and 0.6-110 ng mL(-1), respectively. The proposed method was successfully applied for the determination of the drugs in pharmaceutical preparations, urine, and plasma samples.

  8. On the distribution of uranium in hair: Non-destructive analysis using synchrotron radiation induced X-ray fluorescence microprobe techniques

    NASA Astrophysics Data System (ADS)

    Israelsson, A.; Eriksson, M.; Pettersson, H. B. L.

    2015-06-01

    In the present study the distribution of uranium in single human hair shafts has been evaluated using two synchrotron radiation (SR) based micro X-ray fluorescence techniques; SR μ-XRF and confocal SR μ-XRF. The hair shafts originated from persons that have been exposed to elevated uranium concentrations. Two different groups have been studied, i) workers at a nuclear fuel fabrication factory, exposed mainly by inhalation and ii) owners of drilled bedrock wells exposed by ingestion of water. The measurements were carried out on the FLUO beamline at the synchrotron radiation facility ANKA, Karlsruhe. The experiment was optimized to detect U with a beam size of 6.8 μm × 3 μm beam focus allowing detection down to ppb levels of U in 10 s (SR μ-XRF setup) and 70 s (SR confocal μ-XRF setup) measurements. It was found that the uranium was present in a 10-15 μm peripheral layer of the hair shafts for both groups studied. Furthermore, potential external hair contamination was studied by scanning of unwashed hair shafts from the workers. Sites of very high uranium signal were identified as particles containing uranium. Such particles, were also seen in complementary analyses using variable pressure electron microscope coupled with energy dispersive X-ray analyzer (ESEM-EDX). However, the particles were not visible in washed hair shafts. These findings can further increase the understanding of uranium excretion in hair and its potential use as a biomonitor.

  9. Stardust Interstellar Preliminary Examination VII: Synchrotron X-Ray Fluorescence Analysis of Six Stardust Interstellar Candidates Measured with the Advanced Photon Source 2-ID-D Microprobe

    NASA Technical Reports Server (NTRS)

    Allen, Carlton C.; Anderson, David; Bastien, Ron K.; Brenker, Frank E.; Flynn, George J.; Frank, David; Gainsforth, Zack; Sandford, Scott A.; Simionovici, Alexandre S.; Zolensky, Michael E.

    2014-01-01

    The NASA Stardust spacecraft exposed an aerogel collector to the interstellar dust passing through the solar system. We performed X-ray fluorescence element mapping and abundance measurements, for elements 19 < or = Z < or = 30, on six "interstellar candidates," potential interstellar impacts identified by Stardust@Home and extracted for analyses in picokeystones. One, I1044,3,33, showed no element hot-spots within the designated search area. However, we identified a nearby surface feature, consistent with the impact of a weak, high-speed particle having an approximately chondritic (CI) element abundance pattern, except for factor-of-ten enrichments in K and Zn and an S depletion. This hot-spot, containing approximately 10 fg of Fe, corresponds to an approximately 350 nm chondritic particle, small enough to be missed by Stardust@Home, indicating that other techniques may be necessary to identify all interstellar candidates. Only one interstellar candidate, I1004,1,2, showed a track. The terminal particle has large enrichments in S, Ti, Cr, Mn, Ni, Cu, and Zn relative to Fe-normalized CI values. It has high Al/Fe, but does not match the Ni/Fe range measured for samples of Al-deck material from the Stardust sample return capsule, which was within the field-of-view of the interstellar collector. A third interstellar candidate, I1075,1,25, showed an Al-rich surface feature that has a composition generally consistent with the Al-deck material, suggesting that it is a secondary particle. The other three interstellar candidates, I1001,1,16, I1001,2,17, and I1044,2,32, showed no impact features or tracks, but allowed assessment of submicron contamination in this aerogel, including Fe hot-spots having CI-like Ni/Fe ratios, complicating the search for CI-like interstellar/interplanetary dust.

  10. A CMOS In-Pixel CTIA High Sensitivity Fluorescence Imager

    PubMed Central

    Murari, Kartikeya; Etienne-Cummings, Ralph; Thakor, Nitish; Cauwenberghs, Gert

    2012-01-01

    Traditionally, charge coupled device (CCD) based image sensors have held sway over the field of biomedical imaging. Complementary metal oxide semiconductor (CMOS) based imagers so far lack sensitivity leading to poor low-light imaging. Certain applications including our work on animal-mountable systems for imaging in awake and unrestrained rodents require the high sensitivity and image quality of CCDs and the low power consumption, flexibility and compactness of CMOS imagers. We present a 132×124 high sensitivity imager array with a 20.1 μm pixel pitch fabricated in a standard 0.5 μ CMOS process. The chip incorporates n-well/p-sub photodiodes, capacitive transimpedance amplifier (CTIA) based in-pixel amplification, pixel scanners and delta differencing circuits. The 5-transistor all-nMOS pixel interfaces with peripheral pMOS transistors for column-parallel CTIA. At 70 fps, the array has a minimum detectable signal of 4 nW/cm2 at a wavelength of 450 nm while consuming 718 μA from a 3.3 V supply. Peak signal to noise ratio (SNR) was 44 dB at an incident intensity of 1 μW/cm2. Implementing 4×4 binning allowed the frame rate to be increased to 675 fps. Alternately, sensitivity could be increased to detect about 0.8 nW/cm2 while maintaining 70 fps. The chip was used to image single cell fluorescence at 28 fps with an average SNR of 32 dB. For comparison, a cooled CCD camera imaged the same cell at 20 fps with an average SNR of 33.2 dB under the same illumination while consuming over a watt. PMID:23136624

  11. Metallic Nanomaterials for Sensitivity Enhancement of Fluorescence Detection

    PubMed Central

    Goldys, Ewa M.; Xie, Fang

    2008-01-01

    Utrasensitive detection of trace analytes by fluorescence benefits for fluorescence amplifying substrates. We review here our recent work concerned with understanding of enhancement mechanisms and formation of three such substrates: silver fractals, silver coated gold nanoparticles deposited on glass and fluorescence enhancing gold colloids. PMID:27879741

  12. Positron microprobe at LLNL

    SciTech Connect

    Asoka, P; Howell, R; Stoeffl, W

    1998-11-01

    The electron linac based positron source at Lawrence Livermore National Laboratory (LLNL) provides the world's highest current beam of keV positrons. We are building a positron microprobe that will produce a pulsed, focused positron beam for 3-dimensional scans of defect size and concentration with sub-micron resolution. The widely spaced and intense positron packets from the tungsten moderator at the end of the 100 MeV LLNL linac are captured and trapped in a magnetic bottle. The positrons are then released in 1 ns bunches at a 20 MHz repetition rate. With a three-stage re-moderation we will compress the cm-sized original beam to a 1 micro-meter diameter final spot on the target. The buncher will compress the arrival time of positrons on the target to less than 100 ps. A detector array with up to 60 BaF2 crystals in paired coincidence will measure the annihilation radiation with high efficiency and low background. The energy of the positrons can be varied from less than 1 keV up to 50 keV.

  13. Increased sensitivity of bacterial detection in cerebrospinal fluid by fluorescent staining on low-fluorescence membrane filters.

    PubMed

    Durtschi, Jacob D; Erali, Maria; Bromley, L Kathryn; Herrmann, Mark G; Petti, Cathy A; Smith, Roger E; Voelkerding, Karl V

    2005-09-01

    A membrane-filter-based, fluorescent Gram stain method for bacterial detection in cerebrospinal fluid samples was developed and evaluated as a rapid, sensitive alternative to standard Gram stain protocols. A recently developed, modified version of the aluminium oxide membrane Anopore with low-fluorescence optical properties showed superior performance in this application. Other aspects of the fluorescent Gram stain system that were evaluated include membrane filter selection, strategies to reduce fluorescence fading and the effect of patient blood cells on bacterial detection in the fluorescently stained cerebrospinal fluid samples. The combination of the membrane filter's bacteria-concentrating ability and absolute retention along with high-contrast, fluorescent Gram discriminating dyes enabled rapid bacterial detection and Gram discrimination, with a 1-1.5 order of magnitude increase in the bacterial concentration limit of detection.

  14. Fluorescent Probe Encapsulated in SNAP-Tag Protein Cavity To Eliminate Nonspecific Fluorescence and Increase Detection Sensitivity.

    PubMed

    Zeng, Yan-Syun; Gao, Ruo-Cing; Wu, Ting-Wei; Cho, Chien; Tan, Kui-Thong

    2016-08-17

    Despite the promising improvements made recently on fluorescence probes for the detection of enzymes and reactive small molecules, two fundamental problems remain: weaker fluorescence of many dyes in aqueous buffers and strong nonspecific signals in samples containing high protein levels. In this paper, we introduce a novel fluorescent probe encapsulated in protein cavity (FPEPC) concept as demonstrated by SNAP-tag protein and three environment-sensitive fluorescence probes to overcome these two problems. The probes were constructed by following the current probe design for enzymes and reactive small molecules but with an additional benzylguanine moiety for selective SNAP-tag conjugation. The SNAP-tag conjugated probes achieved quantitative nitroreductase and hydrogen sulfide detection in blood plasma, whereas analyte concentrations were overestimated up to 700-fold when bare fluorescent probes were employed for detection. Furthermore, detection sensitivity was increased dramatically, as our probes displayed 390-fold fluorescence enhancement upon SNAP-tag conjugation, in stark contrast to the weak fluorescence of the free probes in aqueous solutions. Compared with the conventional approaches where fluorescent probes are encapsulated into polymers and nanoparticles, our simple and general approach successfully overcame many key issues such as dye leakage, long preparation steps, inconsistent dye-host ratios, difficulty in constructing in situ in a complex medium, and limited application to detect only small metabolites.

  15. Focusing optics for a synchrotron x radiation microprobe

    SciTech Connect

    Ice, G.E.; Sparks, C.J. Jr.

    1983-01-01

    We propose two constant deviation and energy-tunable fluorescent microprobe optical designs which efficiently use x rays available from ending magnets and insertion devices of synchrotron radiation sources. The simpler system consists of a cylindrically bent multilayer to focus the vertical opening angle by in-plane scattering, a fixed radius cylindrically curved multilayer which sagittally focuses the horizontal divergence, and a pinhole to further reduce the beam to microprobe dimensions. A more versatile system has a pair of flat nondispersively arranged diffracting optics followed by crossed elliptical mirrors. These nondispersive combinations can produce a fixed-exit beam. We compare the relative intensity with other optical systems.

  16. Trace Element Zoning and Incipient Metamictization in a Lunar Zircon: Application of Three Microprobe Techniques

    NASA Technical Reports Server (NTRS)

    Wopenka, Brigitte; Jollife, Bradley L.; Zinner, Ernst; Kremser, Daniel T.

    1996-01-01

    We have determined major (Si, Zr, Hf), minor (Al, Y, Fe, P), and trace element (Ca, Sc, Ti, Ba, REE, Th, U) concentrations and Raman spectra of a zoned, 200 microns zircon grain in lunar sample 14161,7069, a quartz monzodiorite breccia collected at the Apollo 14 site. Analyses were obtained on a thin section in situ with an ion microprobe, an electron microprobe, and a laser Raman microprobe. The zircon grain is optically zoned in birefringence, a reflection of variable (incomplete) metamictization resulting from zo- nation in U and Th concentrations. Variations in the concentrations of U and Th correlate strongly with those of other high-field-strength trace elements and with changes in Raman spectral parameters. Concentrations of U and Th range from 21 to 55 ppm and 6 to 31 ppm, respectively, and correlate with lower Raman peak intensities, wider Raman peaks, and shifted Si-O peak positions. Concentrations of heavy rare earth elements range over a factor of three to four and correlate with intensities of fluorescence peaks. Correlated variations in trace element concentrations reflect the original magmatic differentiation of the parental melt approx. 4 b.y. ago. Degradation of the zircon structure, as reflected by the observed Raman spectral parameters, has occurred in this sample over a range of alpha-decay event dose from approx. 5.2 x 10(exp 14) to 1.4 x 10(exp 15) decay events per milligram of zircon, as calculated from the U and Th concentrations. This dose is well below the approx. 10(exp 16) events per milligram cumulative dose that causes complete metamictization and indicates that laser Raman microprobe spectroscopy is an analytical technique that is very sensitive to the radiation-induced damage in zircon.

  17. A microenvironment-sensitive fluorescent pyrimidine ribonucleoside analogue: synthesis, enzymatic incorporation, and fluorescence detection of a DNA abasic site.

    PubMed

    Tanpure, Arun A; Srivatsan, Seergazhi G

    2011-11-04

    Base-modified fluorescent ribonucleoside-analogue probes are valuable tools in monitoring RNA structure and function because they closely resemble the structure of natural nucleobases. Especially, 2-aminopurine, a highly environment-sensitive adenosine analogue, is the most extensively utilized fluorescent nucleoside analogue. However, only a few isosteric pyrimidine ribonucleoside analogues that are suitable for probing the structure and recognition properties of RNA molecules are available. Herein, we describe the synthesis and photophysical characterization of a small series of base-modified pyrimidine ribonucleoside analogues derived from tagging indole, N-methylindole, and benzofuran onto the 5-position of uracil. One of the analogues, based on a 5-(benzofuran-2-yl)pyrimidine core, shows emission in the visible region with a reasonable quantum yield and, importantly, displays excellent solvatochromism. The corresponding triphosphate substrate is effectively incorporated into oligoribonucleotides by T7 RNA polymerase to produce fluorescent oligoribonucleotide constructs. Steady-state and time-resolved spectroscopic studies with fluorescent oligoribonucleotide constructs demonstrate that the fluorescent ribonucleoside photophysically responds to subtle changes in its environment brought about by the interaction of the chromophore with neighboring bases. In particular, the emissive ribonucleoside, if incorporated into an oligoribonucleotide, positively reports the presence of a DNA abasic site with an appreciable enhancement in fluorescence intensity. The straightforward synthesis, amicability to enzymatic incorporation, and sensitivity to changes in the microenvironment highlight the potential of the benzofuran-conjugated pyrimidine ribonucleoside as an efficient fluorescent probe to investigate nucleic acid structure, dynamics, and recognition events.

  18. Electron microprobe mineral analysis guide

    NASA Technical Reports Server (NTRS)

    Brown, R. W.

    1980-01-01

    Electron microprobe mineral analysis guide is a compilation of X-ray tables and spectra recorded from various mineral matrices. Spectra were obtained using electron microprobe, equipped with LiF geared, curved crystal X-ray spectrometers, utilizing typical analytical operating conditions: 15 Kv acceleration potential, 0.02 microampere sample current as measured on a clinopyroxene standard (CP19). Tables and spectra are presented for the majority of elements, fluorine through uranium, occurring in mineral samples from lunar, meteoritic and terrestrial sources. Tables for each element contain relevant analytical information, i.e., analyzing crystal, X-ray peak, background and relative intensity information, X-ray interferences and a section containing notes on the measurement. Originally intended to cover silicates and oxide minerals the tables and spectra have been expanded to cover other mineral phases. Electron microprobe mineral analysis guide is intended as a spectral base to which additional spectra can be added as the analyst encounters new mineral matrices.

  19. Sensitive high resolution ion microprobe (SHRIMP) detrital zircon geochronology provides new evidence for a hidden neoproterozoic foreland basin to the Grenville Orogen in the eastern Midwest, U.S.A

    USGS Publications Warehouse

    Santos, J.O.S.; Hartmann, L.A.; McNaughton, N.J.; Easton, R. M.; Rea, R.G.; Potter, P.E.

    2002-01-01

    A sensitive high resolution ion microprobe (SHRIMP) was used in combination with backscattered electron (BSE) and cathodoluminescence (CL) images to determine the age of detrital zircons from sandstones in the Neoproterozoic Middle Run Formation of the eastern Midwest, United States. Eleven samples from seven drill cores of the upper part of the Middle Run Formation contain detrital zircons ranging in age from 1030 to 1982 Ma (84 analyses), with six distinctive modes at 1.96, 1.63, 1.47, 1.34, 1.15, and 1.08 Ga. This indicates that most, but not all, of the zircon at the top of the Middle Run Formation was derived from the Grenville Orogen. The youngest concordant detrital zircon yields a maximum age of 1048 ?? 22 Ma for the Middle Run Formation, indicating that the formation is younger than ca. 1026 Ma minus the added extra time needed for later uplift, denudation, thrusting, erosion, and transport to southwestern Ohio. Thus, as judged by proximity, composition, thickness, and geochronology, it is a North American equivalent to other Neoproterozoic Grenvillian-derived basins, such as the Torridon Group of Scotland and the Palmeiral Formation of South America. An alternate possibility, although much less likely in our opinion, is that it could be much younger, any time between 1048 ?? 22 Ma and the deposition of the Middle Cambrian Mount Simon Sandstone at about 510 Ma, and still virtually almost all derived from rocks of the Grenville Orogen.

  20. Improved sensitivity to fluorescence for cancer detection in wide-field image-guided neurosurgery

    PubMed Central

    Jermyn, Michael; Gosselin, Yoann; Valdes, Pablo A.; Sibai, Mira; Kolste, Kolbein; Mercier, Jeanne; Angulo, Leticia; Roberts, David W.; Paulsen, Keith D.; Petrecca, Kevin; Daigle, Olivier; Wilson, Brian C.; Leblond, Frederic

    2015-01-01

    In glioma surgery, Protoporphyrin IX (PpIX) fluorescence may identify residual tumor that could be resected while minimizing damage to normal brain. We demonstrate that improved sensitivity for wide-field spectroscopic fluorescence imaging is achieved with minimal disruption to the neurosurgical workflow using an electron-multiplying charge-coupled device (EMCCD) relative to a state-of-the-art CMOS system. In phantom experiments the EMCCD system can detect at least two orders-of-magnitude lower PpIX. Ex vivo tissue imaging on a rat glioma model demonstrates improved fluorescence contrast compared with neurosurgical fluorescence microscope technology, and the fluorescence detection is confirmed with measurements from a clinically-validated spectroscopic probe. Greater PpIX sensitivity in wide-field fluorescence imaging may improve the residual tumor detection during surgery with consequent impact on survival. PMID:26713218

  1. Determination of flumioxazin residue in food samples through a sensitive fluorescent sensor based on click chemistry.

    PubMed

    Lu, Lijun; Yang, Linlin; Cai, Huijian; Zhang, Lan; Lin, Zhenyu; Guo, Longhua; Qiu, Bin; Chen, Guonan

    2014-11-01

    A sensitive and selective fluorescent sensor for flumioxazin was designed based on the formation of strong fluorescence compound (1,2,3-triazole compounds) via the reaction of the alkynyl group in flumioxazin with 3-azido-7-hydroxycoumarin, a weak-fluorescent compound, through the Cu(+)-catalysed azide-alkyne cycloaddition (CuAAC) reaction. The fluorescence increase factor (represented by F/F0) of the system exhibited a good linear relationship with the concentrations of flumioxazin in the range of 0.25-6.0 μg/L with a detection limit of 0.18 μg/L (S/N=3). Also, the proposed fluorescent sensor demonstrated good selectivity for flumioxazin assay even in the presence of high concentration of other pesticides. Based on such high sensitivity and selectivity, the proposed fluorescent sensor has been applied to test the flumioxazin residue in some vegetable and water samples with satisfied results.

  2. Improved sensitivity to fluorescence for cancer detection in wide-field image-guided neurosurgery.

    PubMed

    Jermyn, Michael; Gosselin, Yoann; Valdes, Pablo A; Sibai, Mira; Kolste, Kolbein; Mercier, Jeanne; Angulo, Leticia; Roberts, David W; Paulsen, Keith D; Petrecca, Kevin; Daigle, Olivier; Wilson, Brian C; Leblond, Frederic

    2015-12-01

    In glioma surgery, Protoporphyrin IX (PpIX) fluorescence may identify residual tumor that could be resected while minimizing damage to normal brain. We demonstrate that improved sensitivity for wide-field spectroscopic fluorescence imaging is achieved with minimal disruption to the neurosurgical workflow using an electron-multiplying charge-coupled device (EMCCD) relative to a state-of-the-art CMOS system. In phantom experiments the EMCCD system can detect at least two orders-of-magnitude lower PpIX. Ex vivo tissue imaging on a rat glioma model demonstrates improved fluorescence contrast compared with neurosurgical fluorescence microscope technology, and the fluorescence detection is confirmed with measurements from a clinically-validated spectroscopic probe. Greater PpIX sensitivity in wide-field fluorescence imaging may improve the residual tumor detection during surgery with consequent impact on survival.

  3. Charge-sensitive fluorescent nanosensors created from nanodiamonds.

    PubMed

    Petrakova, V; Rehor, I; Stursa, J; Ledvina, M; Nesladek, M; Cigler, P

    2015-08-07

    We show that fluorescent nanodiamonds (FNDs) are among the few types of nanosensors that enable direct optical reading of noncovalent molecular events. The unique sensing mechanism is based on switching between the negatively charged and neutral states of NV centers which is induced by the interaction of the FND surface with charged molecules.

  4. Mn-Cr relative sensitivity factor in ferromagnesian olivines defined for SIMS measurements with a Cameca ims-1280 ion microprobe: Implications for dating secondary fayalite

    NASA Astrophysics Data System (ADS)

    Doyle, Patricia M.; Jogo, Kaori; Nagashima, Kazuhide; Huss, Gary R.; Krot, Alexander N.

    2016-02-01

    The short-lived radionuclide 53Mn, which decays to 53Cr with a half-life of ∼3.7 Myr, is useful for sequencing objects that formed within the first 20 Myr of Solar System evolution. 53Mn-53Cr relative chronology enables aqueously formed secondary minerals such as fayalite and various carbonates in ordinary and carbonaceous chondrites to be dated, thereby providing chronological constraints on aqueous alteration processes. In situ measurements of Mn-Cr isotope systematics in fayalite by secondary ion mass spectrometry (SIMS) require consideration of the relative sensitivities of the 55Mn+ and 52Cr+ ions, for which a relative sensitivity factor [RSF = (55Mn+/52Cr+)SIMS/(55Mn/52Cr)true] is defined using appropriate standards. In the past, San Carlos olivine (Fa∼10) was commonly used for this purpose, but a growing body of evidence suggests that it is an unsuitable standard for meteoritic fayalite (Fa>90). Natural fayalite also cannot be used as a standard because it contains only trace amounts of chromium, which makes determining a true 55Mn/52Cr ratio and its degree of heterogeneity very difficult. To investigate the dependence of the Mn-Cr RSF on ferromagnesian olivine compositions, we synthesized a suite of compositionally homogeneous Mn,Cr-bearing liquidus-phase ferromagnesian olivines (Fa31-99). Manganese-chromium isotopic measurements of San Carlos olivine and synthesized ferromagnesian olivines using the University of Hawai'i Cameca ims-1280 SIMS show that the RSF for Fa10 is ∼0.9; it increases rapidly between Fa10 and Fa31 and reaches a plateau value of ∼1.5 ± 0.1 for Fa>34. The RSF is time-dependent: it increases during the measurements of olivines with fayalite content <30 and decreases during the measurements of olivines with fayalite content >50. The RSF measured on ferroan olivine (Fa>90) is influenced by pit shape, whereas the RSF measured on magnesian olivine (Fa10) is less sensitive to changes in pit shape. For these reasons, 53Mn-53Cr

  5. Matrix effects on the relative sensitivity factors for manganese and chromium during ion microprobe analysis of carbonate: Implications for early Solar System chronology

    NASA Astrophysics Data System (ADS)

    Steele, Robert C. J.; Heber, Veronika S.; McKeegan, Kevin D.

    2017-03-01

    The short-lived radionuclide 53 Mn decays to 53 Cr providing a relative chronometer for dating the formation of Mn-rich minerals in meteorites. Secondary ion mass spectrometry (SIMS) has been extensively used for in situ dating of meteoritic olivine and carbonate by the 53 Mn-53 Cr system, however a significant analytical challenge has been realising accurate measurements of the Mn/Cr ratio in individual minerals of differing chemical compositions. During SIMS analysis, elements are ionised with differing efficiencies and standard materials are required to calibrate measured ion intensities in terms of relative elemental concentrations. The carbonate system presents a particular analytical difficulty since such standards are not naturally available due to low and variable Cr contents. Here, we utilise ion implantation of Cr into carbonate and other phases to accurately determine the relative sensitivity factors of Mn/Cr during SIMS analysis. We find significant variations in Mn/Cr RSF values among different carbonate minerals that depend systematically on chemical composition and we propose an empirical correction scheme that quantitatively yields an accurate RSF for carbonates of diverse chemical compositions. Correction of SIMS carbonate data for this strong matrix effect may help to reconcile some outstanding problems regarding the timescales of aqueous alteration processes in carbonaceous chondrites. Mn-Cr ages, revised based our new understanding of the matrix effect, are, in general, earlier than previously thought and the duration of carbonate formation is shorter.

  6. Aptamer-Functionalized Fluorescent Silica Nanoparticles for Highly Sensitive Detection of Leukemia Cells

    NASA Astrophysics Data System (ADS)

    Tan, Juntao; Yang, Nuo; Hu, Zixi; Su, Jing; Zhong, Jianhong; Yang, Yang; Yu, Yating; Zhu, Jianmeng; Xue, Dabin; Huang, Yingying; Lai, Zongqiang; Huang, Yong; Lu, Xiaoling; Zhao, Yongxiang

    2016-06-01

    A simple, highly sensitive method to detect leukemia cells has been developed based on aptamer-modified fluorescent silica nanoparticles (FSNPs). In this strategy, the amine-labeled Sgc8 aptamer was conjugated to carboxyl-modified FSNPs via amide coupling between amino and carboxyl groups. Sensitivity and specificity of Sgc8-FSNPs were assessed using flow cytometry and fluorescence microscopy. These results showed that Sgc8-FSNPs detected leukemia cells with high sensitivity and specificity. Aptamer-modified FSNPs hold promise for sensitive and specific detection of leukemia cells. Changing the aptamer may allow the FSNPs to detect other types of cancer cells.

  7. Fluorescent Gold Nanoprobes for the Sensitive and Selective Detection for Hg2+

    NASA Astrophysics Data System (ADS)

    Chai, Fang; Wang, Tingting; Li, Lu; Liu, Haiyan; Zhang, Lingyu; Su, Zhongmin; Wang, Chungang

    2010-11-01

    A simple, cost-effective yet rapid and sensitive sensor for on-site and real-time Hg2+ detection based on bovine serum albumin functionalized fluorescent gold nanoparticles as novel and environmentally friendly fluorescent probes was developed. Using this probe, aqueous Hg2+ can be detected at 0.1 nM in a facile way based on fluorescence quenching. This probe was also applied to determine the Hg2+ in the lake samples, and the results demonstrate low interference and high sensitivity.

  8. DS-2 Mars Microprobe Battery

    NASA Technical Reports Server (NTRS)

    Frank, H.; Kindler, A.; Deligiannis, F.; Davies, E.; Blankevoort, J.; Ratnakumar, B. V.; Surampudi, S.

    1999-01-01

    In January of 1999 the NM DS-2 Mars microprobe will be launched to impact on Mars in December. The technical objectives of the missions are to demonstrate: key technologies, a passive atmospheric entry, highly integrated microelectronics which can withstand both low temperatures and high decelerations, and the capability to conduct in-situ, surface and subsurface science data acquisition. The scientific objectives are to determine if ice is present below the Martian surface, measure the local atmospheric pressure, characterize the thermal properties of the martian subsurface soil, and to estimate the vertical temperature gradient of the Martian soil. The battery requirements are 2-4 cell batteries, with voltage of 6-14 volts, capacity of 550 mAh at 80C, and 2Ah at 25C, shelf life of 2.5 years, an operating temperature of 60C and below, and the ability to withstand shock impact of 80,000 g's. The technical challenges and the approach is reviewed. The Li-SOCL2 system is reviewed, and graphs showing the current and voltage is displayed, along with the voltage over discharge time. The problems encountered during the testing were: (1) impact sensitivity, (2) cracking of the seals, and (3) delay in voltage. A new design resulted in no problems in the impact testing phase. The corrective actions for the seal problems involved: (1) pre weld fill tube, (2) an improved heat sink during case to cover weld and (3) change the seal dimensions to reduce stress. To correct the voltage delay problem the solutions involved: (1) drying the electrodes to reduce contamination by water, (2) assemblage of the cells within a week of electrode manufacture, (3) ensure electrolyte purity, and (4) provide second depassivation pulse after landing. The conclusions on further testing were that the battery can: (1) withstand anticipated shock of up to 80,000 g, (2) meet the discharge profile post shock at Mars temperatures, (3) meet the required self discharge rate and (4) meet environmental

  9. In vivo monitoring of nanosphere onsite delivery using fiber optic microprobe

    NASA Astrophysics Data System (ADS)

    Lo, Leu-Wei; Yang, Chung-Shi

    2005-02-01

    To recognize the information of ischemia-induced blood vessel permeability would be valuable to formulate the drugs for optimal local delivery, we constructed an implantable needle type fiber-optic microprobe for the monitoring of in vivo fluorescent substances in anesthetized rats. This fiber-optic microprobe was composed of coaxial optical fibers and catheterized using a thin wall tubing of stainless steel (~400 um O.D. and ~300 um I.D.). The central fiber, with 100 um core diameter and 20 um cladding, coated with a 30 um layer of gold, was surrounded by 10 fibers with 50 um cores. The central fiber carried the light from the 488 nm Argon laser to the tissue while the surrounding fibers collected the emitted fluorescence to the detector. When the fiber-optic microprobe was placed in the solutions containing various concentrations of fluorescent nanospheres (20 nm), either with or without 10% lipofundin as optical phantom, nanosphere concentration-dependent responses of the fluorescence intensity were observed. The microprobe was then implanted into the liver and the brain of anesthetized rats to monitor the in situ extravasation of pre-administered fluorescent nanospheres from vasculature following the ischemic insults. Both the hepatic and cerebral ischemic insults showed immediate increases of the extracellular 20 nm fluorescent nanospheres. The implantable fiber-optic microprobe constructed in present study provides itself as a minimally-invasive technique capable of investigating the vascular permeability for in vivo nanosphere delivery in both ischemic liver and brain.

  10. Highly sensitive C-reactive protein (CRP) assay using metal-enhanced fluorescence (MEF)

    NASA Astrophysics Data System (ADS)

    Zhang, Yi; Keegan, Gemma L.; Stranik, Ondrej; Brennan-Fournet, Margaret E.; McDonagh, Colette

    2015-07-01

    Fluorescence has been extensively employed in the area of diagnostic immunoassays. A significant enhancement of fluorescence can be achieved when noble metal nanoparticles are placed in close proximity to fluorophores. This effect, referred to as metal-enhanced fluorescence (MEF), has the potential to produce immunoassays with a high sensitivity and a low limit of detection (LOD). In this study, we investigate the fluorescence enhancement effect of two different nanoparticle systems, large spherical silver nanoparticles (AgNPs) and gold edge-coated triangular silver nanoplates, and both systems were evaluated for MEF. The extinction properties and electric field enhancement of both systems were modeled, and the optimum system, spherical AgNPs, was used in a sandwich immunoassay for human C-reactive protein with a red fluorescent dye label. A significant enhancement in the fluorescence was observed, which corresponded to an LOD improvement of 19-fold compared to a control assay without AgNPs.

  11. Fluorescence probe for the convenient and sensitive detection of ascorbic acid

    PubMed Central

    Matsuoka, Yuta; Yamato, Mayumi; Yamada, Ken-ichi

    2016-01-01

    Ascorbic acid is an important antioxidant that plays an essential role in the biosynthesis of numerous bioactive substances. The detection of ascorbic acid has traditionally been achieved using high-performance liquid chromatography and absorption spectrophotometry assays. However, the development of fluorescence probes for this purpose is highly desired because they provide a much more convenient and highly sensitive technique for the detection of this material. OFF-ON-type fluorescent probes have been developed for the detection of non-fluorescent compounds. Photo-induced electron transfer and fluorescence resonance energy transfer are the two main fluorescence quenching mechanisms for the detection of ascorbic acid, and several fluorescence probes have been reported based on redox-responsive metals and quantum dots. Profluorescent nitroxide compounds have also been developed as non-metal organic fluorescence probes for ascorbic acid. These nitroxide systems have a stable unpaired electron and can therefore react with ascorbic acid and a strong fluorescence quencher. Furthermore, recent synthetic advances have allowed for the synthesis of α-substituted nitroxides with varying levels of reactivity towards ascorbic acid. In this review, we have discussed the design strategies used for the preparation of fluorescent probes for ascorbic acid, with particular emphasis on profluorescent nitroxides, which are unique radical-based redox-active fluorescent probes. PMID:26798193

  12. A simple and effective strategy to increase the sensitivity of fluorescence probes in living cells.

    PubMed

    Izumi, Saki; Urano, Yasuteru; Hanaoka, Kenjiro; Terai, Takuya; Nagano, Tetsuo

    2009-07-29

    Noninvasive visualization and investigation of interactions among proteins, biomolecules, and enzymes in living cells is an important goal for biologists, and fluorescence probes are powerful tools for this purpose. Because many target molecules are present in only trace amounts, high sensitivity is very important, and it is common to improve the sensitivity of fluorescence probes by focusing on high reaction velocity, K(d). (Gee, K. R.; Archer, E. A.; Lapham, L. A.; Leonard, M. E.; Zhou, Z.; Bingham, J.; Diwu, Z. Bioorg. Med. Chem. Lett. 2000, 10, 1515-1518.) So far, we have designed and synthesized various highly sensitive fluorescence probes based on the above concepts. (Gabe, Y.; Urano, Y.; Kikuchi, K.; Kojima, H.; Nagano, T. J. Am. Chem. Soc. 2004, 126, 3357-3367. Komatsu, K.; Urano, Y.; Kojima, H.; Nagano, T. J. Am. Chem. Soc. 2007, 129, 13447-13454.) Nevertheless, they were sometimes insufficiently sensitive to detect biomolecules in living cells, despite high chemical sensitivity in cuvette. In this report, we suggest a new approach to increase the sensitivity of fluorescence probes, focusing on their intracellular retention. Since calcein is well-retained, we investigated its structural, chemical, and optical characteristics and found that the iminodiacetic acid group (IAG) is a key structure for the intracellular retention. We next designed and synthesized novel fluorescence probes containing IAGs. They showed superior intracellular retention, making it possible to visualize low concentrations of target molecules that would be difficult to observe with conventional probes and permitting long-term observation in living cells. Improvement of intracellular retention of fluorescence probes holds great promise as a strategy for developing a wide range of highly sensitive probes for studies on various biological phenomena.

  13. An ESIPT-based fluorescent probe for sensitive detection of hydrazine in aqueous solution.

    PubMed

    Zhou, Ji; Shi, Ruiyan; Liu, Jianxu; Wang, Rui; Xu, Yufang; Qian, Xuhong

    2015-05-21

    A fluorescent probe for sensitive detection of hydrazine based on an ESIPT mechanism and a substitution-cyclization-elimination cascade was developed. After the addition of hydrazine, an approximately 50-fold enhancement in fluorescence intensity at 465 nm was observed and the subsequent decrease at 375 nm was observed in 10 min with a detection limit of 0.147 μM. We also detected hydrazine in HeLa cells successfully.

  14. Organic light-emitting device with a phosphor-sensitized fluorescent emission layer

    DOEpatents

    Forrest, Stephen; Kanno, Hiroshi

    2009-08-25

    The present invention relates to organic light emitting devices (OLEDs), and more specifically to OLEDS that emit light using a combination of fluorescent emitters and phosphorescent emitters. The emissive region of the devices of the present invention comprise at least one phosphor-sensitized layer which has a combined emission from a phosphorescent emitter and a fluorescent emitter. In preferred embodiments, the invention relates to white-emitting OLEDS (WOLEDs).

  15. A Sensitive Ratiometric Fluorescent Sensor for Zinc(II) with High Selectivity

    PubMed Central

    Lv, Yuanyuan; Cao, Mingda; Li, Jiakai; Wang, Junbo

    2013-01-01

    A new fluorescent Zn2+ chemosensor (P1) based on a functionalized porphyrin was synthesized and characterized. P1 displayed dramatic ratiometric variations in absorption and fluorescent emission spectra upon exposure to Zn2+ due to the formation of a 1:1 Zn2+/P1 complex. The sensor also exhibited high selectivity and sensitivity toward Zn2+ over other common metal ions in the physiological pH range with a detection limit of 1.8 μM. The sensor showed fast response times and excellent reproducibility, thus confirming its potential applicability as a fluorescent sensor for Zn2+ sensing. PMID:23467028

  16. Hollow-core photonic crystal fiber probe for remote fluorescence sensing with single molecule sensitivity.

    PubMed

    Ghenuche, Petru; Rigneault, Hervé; Wenger, Jérôme

    2012-12-17

    Current optical fiber probes for fluorescence spectroscopy struggle with large luminescence background and low detection sensitivities that challenge the detection of fluorescent molecules at sub-micromolar concentration. Here we report the demonstration of a hollow-core photonic crystal fiber (HC-PCF) probe for remote fluorescence sensing with single molecule sensitivity down to nanomolar concentrations, where both the excitation and fluorescence beams are counter-propagating through the same fiber. A 20 μm polystyrene microsphere is used to efficiently excite and collect the fluorescence from the sample solution thanks to a photonic nanojet effect. Compared to earlier work with silica fibers, the new HC-PCF-microsphere probe achieves a 200x improvement of the signal-to-noise ratio for a single molecule detection event, and a 1000x reduction of the minimum detectable concentration. The device is implemented with fluorescence correlation spectroscopy to distinguish between molecules of similar fluorescence spectra based on the analysis of their translational diffusion properties, and provides similar performance as conventional confocal microscopes.

  17. A novel high-sensitive miniaturized optical system for fluorescence detection

    NASA Astrophysics Data System (ADS)

    Yao, Mingjin; Fang, Ji

    2011-03-01

    This paper presents a novel, high sensitive and miniaturized fluorescence detection system which integrated a LED light source, all necessary optical components and a photodiode with preamplifier into one package about 2 cm x 2 cm x 2 cm especially for the applications of lab-on-a-chip, portable bio-detection system and point-of-care diagnostic system. The prototype has been tested using the fluorescence dye 5-Carboxyfluorescein (5-FAM) dissolved into solvent DMSO (Dimethyl Sulfoxide) and diluted with DI water as the testing solution samples. Resolution approximation method is accepted to evaluate the sensitivity. The testing results prove a remarkable sensitivity at pico-scale molar, around 1.08 pM/L, which should meet the most of bio-detection requirements. This cost-effective detection system can be widely integrated to the portable device and system for fluorescent detection in biological, chemical, medical, point-of-care applications.

  18. High current pulsed positron microprobe

    SciTech Connect

    Howell, R.H.; Stoeffl, W.; Kumar, A.; Sterne, P.A.; Cowan, T.E.; Hartley, J.

    1997-05-01

    We are developing a low energy, microscopically focused, pulsed positron beam for defect analysis by positron lifetime spectroscopy to provide a new defect analysis capability at the 10{sup 10} e{sup +}s{sup -l} beam at the Lawrence Livermore National Laboratory electron linac. When completed, the pulsed positron microprobe will enable defect specific, 3-dimensional maps of defect concentrations with sub-micron resolution of defect location. By coupling these data with first principles calculations of defect specific positron lifetimes and positron implantation profiles we will both map the identity and concentration of defect distributions.

  19. Temperature-dependent fluorescence lifetime of a fluorescent polymeric thermometer, poly(N-isopropylacrylamide), labeled by polarity and hydrogen bonding sensitive 4-sulfamoyl-7-aminobenzofurazan.

    PubMed

    Gota, Chie; Uchiyama, Seiichi; Yoshihara, Toshitada; Tobita, Seiji; Ohwada, Tomohiko

    2008-03-13

    Fluorescent molecular thermometers showing temperature-dependent fluorescence lifetimes enable thermal mapping of small spaces such as a microchannel and a living cell. We report the temperature-dependent fluorescence lifetimes of poly(NIPAM-co-DBD-AA), which is a random copolymer of N-isopropylacrylamide (NIPAM) and an environment-sensitive fluorescent monomer (DBD-AA) containing a 4-sulfamoyl-7-aminobenzofurazan structure. The average fluorescence lifetime of poly(NIPAM-co-DBD-AA) in aqueous solution increased from 4.22 to 14.1 ns with increasing temperature from 30 to 35 degrees C. This drastic change in fluorescence lifetime (27% increase per 1 degrees C) is the sharpest ever reported. Concentration independency, one of the advantages of fluorescence lifetime measurements, was seen in average fluorescence lifetime (13.7 +/- 0.18 ns) of poly(NIPAM-co-DBD-AA) at 33 degrees C over a wide concentration range (0.005-1 w/v%). With increasing temperature, polyNIPAM units in poly(NIPAM-co-DBD-AA) change their structure from an extended form to a globular form, providing apolar and aprotic environments to the fluorescent DBD-AA units. Consequently, the environment-sensitive DBD-AA units translate the local environmental changes into the extension of the fluorescence lifetime. This role of the DBD-AA units was revealed by a study of solvent effects on fluorescence lifetime of a model environment-sensitive fluorophore.

  20. Sensitivity of laser-induced upconversion fluorescence dynamics to exciting wavelength in Er3+-doped YAG

    NASA Astrophysics Data System (ADS)

    Du, S.; Jiang, L.; Zhang, W.; Gong, W.; Fu, Z.; Dai, Z.

    2011-06-01

    A novel sensitivity of laser-induced upconversion (UC) fluorescence dynamics to the exciting wavelength in Er3+:YAG crystal has been observed. The sensitivities to exciting wavelength variations are only 0.02 and 0.06 nm for 2 G 9/2 and 2 P 3/2 UC luminescence, respectively. The observation of this sensitivity reveals that when a certain level is populated by more than one UC mechanisms, a precise determination of suitable exciting wavelength is crucial because the competition between different UC mechanisms has a sensitive variation with exciting wavelength in near-resonant range.

  1. Microscale structure fabrication using microprobe

    NASA Astrophysics Data System (ADS)

    Shinya, Norio; Konno, Takeshi; Egashira, Mitsuru

    1996-05-01

    Using a tungsten micro-probe with a tip of 2 micrometers radius, fine metallic powder particles could be manipulated one by one. By applying low voltage (about 10 V) between the probe and a metallic substrate, the powder particle on the substrate was adsorbed to the tip of probe easily, and by cutting off the voltage the powder particle was desorbed from the tip. Therefore it is possible to arrange powder particles as designed by controlling the voltage and movement of the probe. In addition to the powder particle manipulation, powder particles welding was studied. The tungsten micro-probe was contacted with the powder particle on the metallic substrate, and high voltage (about 10 kV) was applied between the probe and the substrate. It was observed that the glow discharge was caused between the powder particle and the substrate. The contacting parts of the powder particle and the substrate were melted and welded each other. By the manipulation and the welding, micro-structures composed of fine powder particles (about 60 micrometers ) were constructed. Powder particle towers and a micro- actuator were fabricated by way of trial. The results demonstrated the potential of the micro- probe assembly for the fabrication of electronic devices, micromachines and intelligent materials.

  2. Molecular imprinting ratiometric fluorescence sensor for highly selective and sensitive detection of phycocyanin.

    PubMed

    Wang, Xiaoyan; Yu, Jialuo; Kang, Qi; Shen, Dazhong; Li, Jinhua; Chen, Lingxin

    2016-03-15

    A facile strategy was developed to prepare molecular imprinting ratiometric fluorescence sensor for highly selective and sensitive detection of phycocyanin (PC) based on fluorescence resonance energy transfer (FRET), via a sol-gel polymerization process using nitrobenzoxadiazole (NBD) as fluorescent signal source. The ratio of two fluorescence peak emission intensities of NBD and PC was utilized to determine the concentration of PC, which could effectively reduce the background interference and fluctuation of diverse conditions. As a result, this sensor obtained high sensitivity with a low detection limit of 0.14 nM within 6 min, and excellent recognition specificity for PC over its analogues with a high imprinting factor of 9.1. Furthermore, the sensor attained high recoveries in the range of 93.8-110.2% at three spiking levels of PC, with precisions below 4.7% in seawater and lake water samples. The developed sensor strategy demonstrated simplicity, reliability, rapidity, high selectivity and high sensitivity, proving to be a feasible way to develop high efficient fluorescence sensors and thus potentially applicable for ultratrace analysis of complicated matrices.

  3. Hybridization chain reaction amplification for highly sensitive fluorescence detection of DNA with dextran coated microarrays.

    PubMed

    Chao, Jie; Li, Zhenhua; Li, Jing; Peng, Hongzhen; Su, Shao; Li, Qian; Zhu, Changfeng; Zuo, Xiaolei; Song, Shiping; Wang, Lianhui; Wang, Lihua

    2016-07-15

    Microarrays of biomolecules hold great promise in the fields of genomics, proteomics, and clinical assays on account of their remarkably parallel and high-throughput assay capability. However, the fluorescence detection used in most conventional DNA microarrays is still limited by sensitivity. In this study, we have demonstrated a novel universal and highly sensitive platform for fluorescent detection of sequence specific DNA at the femtomolar level by combining dextran-coated microarrays with hybridization chain reaction (HCR) signal amplification. Three-dimensional dextran matrix was covalently coated on glass surface as the scaffold to immobilize DNA recognition probes to increase the surface binding capacity and accessibility. DNA nanowire tentacles were formed on the matrix surface for efficient signal amplification by capturing multiple fluorescent molecules in a highly ordered way. By quantifying microscopic fluorescent signals, the synergetic effects of dextran and HCR greatly improved sensitivity of DNA microarrays, with a detection limit of 10fM (1×10(5) molecules). This detection assay could recognize one-base mismatch with fluorescence signals dropped down to ~20%. This cost-effective microarray platform also worked well with samples in serum and thus shows great potential for clinical diagnosis.

  4. Upconversion fluorescence metal-organic frameworks thermo-sensitive imprinted polymer for enrichment and sensing protein.

    PubMed

    Guo, Ting; Deng, Qiliang; Fang, Guozhen; Gu, Dahai; Yang, Yukun; Wang, Shuo

    2016-05-15

    A novel fluorescence material with thermo-sensitive for the enrichment and sensing of protein was successfully prepared by combining molecular imprinting technology with upconversion nanoparticles (UCNPs) and metal-organic frameworks (MOFs). Herein, the UCNPs acted as signal reporter for composite materials because of its excellent fluorescence property and chemical stability. MOFs were introduced to molecularly imprinted polymer (MIP) due to its high specific surface area which increases the rate of mass transfer relative to that of traditional bulk MIP. The thermo-sensitive imprinted material which allows for swelling and shrinking with response to temperature changes was prepared by choosing Bovine hemoglobin (BHB) as the template, N-isopropyl acrylamide (NIPAAM) as the temperature-sensitive functional monomer and N,N-methylenebisacrylamide (MBA) as the cross-linker. The recognition characterizations of imprinted material-coated UCNPs/MOFs (UCNPs/MOFs/MIP) were evaluated, and the results showed that the fluorescence intensity of UCNPs/MOFs/MIP reduced gradually with the increase of BHB concentration. The fluorescence material was response to the temperature. The adsorption capacity was as much as 167.6 mg/g at 28°C and 101.2mg/g at 44°C, which was higher than that of traditional MIP. Therefore, this new fluorescence material for enrichment and sensing protein is very promising for future applications.

  5. A reliable and sensitive bead-based fluorescence assay for identification of nucleic acid sequences

    NASA Astrophysics Data System (ADS)

    Klamp, Tobias; Yahiatène, Idir; Lampe, André; Schüttpelz, Mark; Sauer, Markus

    2011-03-01

    The sensitive and rapid detection of pathogenic DNA is of tremendous importance in the field of diagnostics. We demonstrate the ability of detecting and quantifying single- and double-stranded pathogenic DNA with picomolar sensitivity in a bead-based fluorescence assay. Selecting appropriate capturing and detection sequences enables rapid (2 h) and reliable DNA quantification. We show that synthetic sequences of S. pneumoniae and M. luteus can be quantified in very small sample volumes (20 μL) across a linear detection range over four orders of magnitude from 1 nM to 1 pM, using a miniaturized wide-field fluorescence microscope without amplification steps. The method offers single molecule detection sensitivity without using complex setups and thus volunteers as simple, robust, and reliable method for the sensitive detection of DNA and RNA sequences.

  6. Fluorescent-conjugated polymer superquenching facilitates highly sensitive detection of proteases

    NASA Astrophysics Data System (ADS)

    Kumaraswamy, Sriram; Bergstedt, Troy; Shi, Xiaobo; Rininsland, Frauke; Kushon, Stuart; Xia, Wensheng; Ley, Kevin; Achyuthan, Komandoor; McBranch, Duncan; Whitten, David

    2004-05-01

    Sensor formats have been developed for detecting the activity of proteolytic enzymes based on fluorescent conjugated polymer superquenching. These sensors employ a reactive peptide sequence within a tether linking a quencher to a biotin. The peptide binds to sensors containing colocated biotin-binding protein and fluorescent polymer by means of biotin-biotin binding protein interactions, resulting in a strong quenching of polymer fluorescence. Enzyme-mediated cleavage of the peptide results in a reversal of the fluorescence quenching. These assays for protease activity are simple, sensitive, fast, and have the specificity required for screening chemical libraries for novel protease inhibitors in a high-throughput screening assay environment. These assays have been demonstrated for enterokinase, caspase-3/7, and -secretase.

  7. Graphene oxide-based fluorescent sensor for sensitive turn-on detection of sinapine

    NASA Astrophysics Data System (ADS)

    Xiang, Xia; Han, Ling; Zhang, Zhen; Huang, Fenghong

    2017-03-01

    Sinapine (SP) has attracted extensive attention in food and medical field because it is a crucial component in many plants with unique physiological activities. Here, we reported a fluorescent sensor based on the unique properties of graphene oxide (GO) for rapid, sensitive detection of SP concentration. In this strategy, GO has stronger affinity toward SP and can be also served as a quencher for DNA fluorescence probe. The fluorescence can make a recovery by the addition of SP, which can competitively desorb probe from the surface of GO, and the SP can thus be monitored by recording the fluorescence change. Because of the low background assays and high quenching ability offered by GO, the developed method provides a great potential for plant-derived molecules research.

  8. Oligonucleotide-stabilized fluorescent silver nanoclusters for the specific and sensitive detection of biotin.

    PubMed

    Xiong, Xiaoli; Tang, Yan; Zhao, Jingjin; Zhao, Shulin

    2016-02-21

    A novel biotin fluorescent probe based on oligonucleotide-stabilized silver nanoclusters (DNA-AgNCs) was synthesized by employing a biotinylated cytosine-rich sequence as a synthesized template. The fluorescence properties of the DNA-AgNCs are related to the modified position of the DNA. When biotin is linked to the middle thymine base of the DNA sequence, the DNA-AgNCs emit the strongest fluorescence. Moreover, the stability of the DNA-AgNCs was affected by avidin through biotin-avidin binding, quenching the fluorescence of the DNA-AgNCs. In contrast, if free biotin is further introduced into this system, the quenching is apparently weakened by competition, leading to the restoration of fluorescence. This phenomenon can be utilized for the detection of biotin. Under the optimal conditions, the fluorescence recovery is linearly proportional to the concentration of biotin in the range of 10 nM-1.0 μM with a detection limit of 6.0 nM. This DNA-AgNCs probe with excellent fluorescent properties is sensitive and selective for the detection of biotin and has been applied for the determination of biotin in wheat flour.

  9. Highly sensitive fluorescence detection of avidin/streptavidin with an optical interference mirror slide.

    PubMed

    Yasuda, Mitsuru; Akimoto, Takuo

    2012-01-01

    This paper presents highly sensitive fluorescence detections of avidin and streptavidin using an optical interference mirror (OIM) slide consisting of a plane mirror covered with an optical interference layer. Compared with a common glass slide, the OIM slide can enhance the fluorescence from a dye by more than 100-fold. We fabricated an OIM slide by depositing an optical interference layer of Al(2)O(3) on an Ag mirror. To enhance the fluorescence maximally, the optimal thickness of the Al(2)O(3) layer was estimated from optical interference theory. For detections of protein, avidin/streptavidin labeled with fluorescein, Cy3, and Cy5 were detected with biotin immobilized on an OIM slide with the optimal Al(2)O(3) thickness. We achieved a sensitivity improvement of more than 50-fold, comparing with a glass slide. Such a high degree of improvement would be a significant contribution to further progress in biomedical research and medical diagnostics.

  10. Estimation of Eu(3+) in bulk uranium by ligand sensitized fluorescence in dimethyl sulphoxide.

    PubMed

    Maji, S; Kumar, Satendra; Sankaran, K

    2014-12-10

    Ligand sensitized fluorescence of europium ion using thenoyltrifluoroacetone (TTA) as a sensitizing ligand and dimethyl sulphoxide (DMSO) as a solvent is studied for the first time. TTA ligand enhances the fluorescence of Eu(3+) by a factor of 40000 in DMSO. Linearity is obtained for a concentration range of 0.076-7.6ng/mL of Eu(3+) with a detection limit of 7.6pg/mL. The quenching of Eu(3+)-TTA fluorescence by uranium matrix was studied in different solvents and found to be less in DMSO. Consequently, estimation of Eu(3+) in a large excess of uranium becomes a possibility without the need to separate uranium from the solution, which has been demonstrated in this paper. Satisfactory results are obtained when Eu(3+) is present at a concentration of 0.6μg/g in uranium.

  11. Glycerin suppression of fluorescence self-quenching and improvement of heterogeneous fluoroimmunoassay sensitivity.

    PubMed

    Petrou, Panagiota S; Mastichiadis, Christos; Christofidis, Ion; Kakabakos, Sotirios E

    2007-01-15

    Fluorescent labels find wide application in immunoassays and immunosensors as well as in protein and DNA chips. However, the use of fluorescent labels in applications requiring high detection sensitivity is limited by fluorescence self-quenching observed when a relatively high number of fluorescent compounds is introduced in the recognition molecule. Here we describe a simple method that suppresses effectively fluorescence self-quenching observed when highly labeled antibodies are used as labels in immunoassays. This was achieved by treating the microtitration wells after the completion of the immunoassay with a glycerin solution followed by 15-min incubation of the emptied wells at 37 degrees C. The remedial action of this method on self-quenching was studied through a noncompetitive immunofluorometric assay for rabbit gamma-globulins employing a sheep anti-rabbit gamma-globulin antibody labeled with fluorescein at molar ratios ranging from 1.0 to 17.4. The glycerin/thermal treatment increased the fluorescence signal measured directly onto the solid surface by 9.2-117% for the antibodies with molar ratios of 1.0-17.4, compared with the values obtained prior to treatment. Furthermore, fluorescence self-quenching was completely removed for labeling ratios up to 14.0. The assay sensitivity was improved 2-4 times by the glycerin/thermal treatment when heavily fluoresceinated antibodies are used as labels (molar ratio >/=5.6). The proposed method resulted also in increased fluorescence signals when labels other than fluorescein were used and improved considerably the detection of protein spots on silicon dies.

  12. Sensitive fluorescence response of ZnSe(S) quantum dots: an efficient fluorescence probe

    NASA Astrophysics Data System (ADS)

    Saikia, K.; Deb, P.; Kalita, E.

    2013-06-01

    An efficient fluorescence probe based on ZnSe(S) alloyed quantum dots (QDs) has been reported here. The alloyed QDs were prepared through an aqueous route, where 3-mercaptopropionic acid (MPA) was employed as the effective precursor for both the sulfur source and stabilizer in the development of the alloyed system. Five-fold quantum yield (QY) enhancement was obtained for the ZnSe(S) QDs compared to the ZnSe QDs, formed in the initial stage of the refluxing process. The ultimate alloyed systems retained their high biocompatibility characteristics similar to the conventional ZnSe QDs. The photoluminescence of the ZnSe(S) QDs showed pH dependence, which was also evidenced in mammalian lymphocyte cells suspended in biological buffer over a wide pH range of 4.00-12.00. These characteristics make our prepared ZnSe(S) an efficient system for development of cell tracking, monitoring and sensing intracellular nanoprobes and devices.

  13. A fluorescent acrylamide-type monomer bearing an environment-sensitive methoxybenzocoumarin structure for the development of functional polymeric sensors.

    PubMed

    Uchiyama, Seiichi; Remón, Patricia; Pischel, Uwe; Kawamoto, Kyoko; Gota, Chie

    2016-10-05

    A new fluorescent acrylamide-type monomer bearing a hydrogen bonding- and polarity-sensitive benzocoumarin fluorophore was synthesized. The absorption spectra, fluorescence spectra, and fluorescence lifetime of a model compound were measured in ten solvents with different hydrogen-bonding abilities and polarities to investigate the sensitivity of the fluorophore to the surrounding environment. These spectroscopic studies demonstrated that the fluorophore emits stronger fluorescence in more protic, polar environments. A fluorescent polymeric thermometer was prepared from N-isopropylacrylamide and the new fluorescent monomer, and it showed good functionality in aqueous solution (e.g., high sensitivity to temperature changes and high chemical stability), indicating the applicability of the herein developed fluorescent monomer for use in functional sensors.

  14. Microprobe analysis of chlorpromazine pigmentation

    SciTech Connect

    Benning, T.L.; McCormack, K.M.; Ingram, P.; Kaplan, D.L.; Shelburne, J.D.

    1988-10-01

    We describe the histochemical, ultrastructural, and microanalytical features of a skin biopsy specimen obtained from a patient with chlorpromazine pigmentation. Golden-brown pigment granules were present in the dermis, predominantly in a perivascular arrangement. The granules stained positively with the Fontana-Masson stain for silver-reducing substances and negatively with Perl's stain for iron. Electron microscopy revealed dense inclusion bodies in dermal histiocytes, pericytes, endothelial cells, and Schwann cells, as well as lying free in the extracellular matrix. These ''chlorpromazine bodies'' were quite dense even in unosmicated, unstained ultrathin sections, indicating that the pigmentation is related, at least in part, to the inclusions. Microprobe analysis of the chlorpromazine bodies revealed a striking peak for sulfur, which strongly suggests the presence of the drug or its metabolite within these inclusions.

  15. Quantum dots based mesoporous structured imprinting microspheres for the sensitive fluorescent detection of phycocyanin.

    PubMed

    Zhang, Zhong; Li, Jinhua; Wang, Xiaoyan; Shen, Dazhong; Chen, Lingxin

    2015-05-06

    Phycocyanin with important physiological/environmental significance has attracted increasing attention; versatile molecularly imprinted polymers (MIPs) have been applied to diverse species, but protein imprinting is still quite difficult. Herein, using phycocyanin as template via a sol-gel process, we developed a novel fluorescent probe for specific recognition and sensitive detection of phycocyanin by quantum dots (QDs) based mesoporous structured imprinting microspheres (SiO2@QDs@ms-MIPs), obeying electron-transfer-induced fluorescence quenching mechanism. When phycocyanin was present, a Meisenheimer complex would be produced between phycocyanin and primary amino groups of QDs surface, and then the photoluminescent energy of QDs would be transferred to the complex, leading to the fluorescence quenching of QDs. As a result, the fluorescent intensity of the SiO2@QDs@ms-MIPs was significantly decreased within 8 min, and accordingly a favorable linearity within 0.02-0.8 μM and a high detectability of 5.9 nM were presented. Excellent recognition specificity for phycocyanin over its analogues was displayed, with a high imprinting factor of 4.72. Furthermore, the validated probe strategy was successfully applied to seawater and lake water sample analysis, and high recoveries in the range of 94.0-105.0% were attained at three spiking levels of phycocyanin, with precisions below 5.3%. The study provided promising perspectives to develop fluorescent probes for convenient, rapid recognition and sensitive detection of trace proteins from complex matrices, and further pushed forward protein imprinting research.

  16. A highly sensitive fluorescence probe for metallothioneins based on tiron-copper complex

    NASA Astrophysics Data System (ADS)

    Xiao, Xilin; Xue, Jinhua; Liao, Lifu; Huang, Mingyang; Zhou, Bin; He, Bo

    2015-06-01

    The fabrication of tiron-copper complex as a novel fluorescence probe for the sensitive directly detection of metallothioneins at nanomolar levels was demonstrated. In Britton-Robinson (B-R) buffer (pH 7.50), the interaction of bis(tiron)copper(II) complex cation [Cu(tiron)2]2+ and metallothioneins enhanced the fluorescence intensity of the system. The fluorescence enhancement at 347 nm was proportional to the concentration of metallothioneins. The mechanism was studied and discussed in terms of the fluorescence spectra. Under the optimal experimental conditions, at 347 nm, there was a linear relationship between the fluorescence intensity and the concentration of the metallothioneins in the range of 8.80 × 10-9-7.70 × 10-7 mol L-1, with a correlation coefficient of r = 0.995 and detection limit 2.60 × 10-9 mol L-1. The relative standard deviation was 0.77% (n = 11), and the average recovery 94.4%. The method proposed was successfully reliable, selective and sensitive in determining of trace metallothioneins in fish visceral organ samples with the results in good agreement with those obtained by HPLC.

  17. Highly selective and sensitive fluorescent paper sensor for nitroaromatic explosive detection.

    PubMed

    Ma, Yingxin; Li, Hao; Peng, Shan; Wang, Leyu

    2012-10-02

    Rapid, sensitive, and selective detection of explosives such as 2,4,6-trinitrotoluene (TNT) and 2,4,6-trinitrophenol (TNP), especially using a facile paper sensor, is in high demand for homeland security and public safety. Although many strategies have been successfully developed for the detection of TNT, it is not easy to differentiate the influence from TNP. Also, few methods were demonstrated for the selective detection of TNP. In this work, via a facile and versatile method, 8-hydroxyquinoline aluminum (Alq(3))-based bluish green fluorescent composite nanospheres were successfully synthesized through self-assembly under vigorous stirring and ultrasonic treatment. These polymer-coated nanocomposites are not only water-stable but also highly luminescent. Based on the dramatic and selective fluorescence quenching of the nanocomposites via adding TNP into the aqueous solution, a sensitive and robust platform was developed for visual detection of TNP in the mixture of nitroaromatics including TNT, 2,4-dinitrotoluene (DNT), and nitrobenzene (NB). Meanwhile, the fluorescence intensity is proportional to the concentration of TNP in the range of 0.05-7.0 μg/mL with the 3σ limit of detection of 32.3 ng/mL. By handwriting or finger printing with TNP solution as ink on the filter paper soaked with the fluorescent nanocomposites, the bluish green fluorescence was instantly and dramatically quenched and the dark patterns were left on the paper. Therefore, a convenient and rapid paper sensor for TNP-selective detection was fabricated.

  18. Highly sensitive detection of superoxide dismutase based on an immunoassay with surface-enhanced fluorescence.

    PubMed

    Yang, Xiaoming; Dou, Yao; Zhu, Shanshan

    2013-06-07

    Herein, a novel highly sensitive enhanced-fluorescence immunoassay for detection of superoxide dismutase (SOD) is established by combining surface-enhanced fluorescence (SEF) with immuno-magnetic separation. Based on a sandwich-type immunoassay, analytes in samples are first captured by magnetic beads coated with a monoclonal antibody and then "sandwiched" by another monoclonal antibody on silver nanoparticles labeled with fluorescein-labeled oligonucleotides in the presence of a magnet. Subsequently, the immune complex is enriched by exposure to a magnetic field. Lastly, the fluorescence intensity is measured according to the number of dissociated fluoresceins. The increased fluorescence intensity permits highly sensitive detection of SOD in a linear range of 10-8 × 10(5) pg mL(-1), with a detection limit of 4 pg mL(-1) at a signal-to-noise ratio of 3. Significantly, this method was validated for detection of SOD in human serum, human urine, and cosmetic samples. Moreover, the reliability and accuracy of results obtained by the enhanced-fluorescence method was confirmed by the analysis of high performance liquid chromatography (HPLC).

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

    NASA Astrophysics Data System (ADS)

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

    2015-07-01

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

  20. Ultra-sensitive and selective Hg{sup 2+} detection based on fluorescent carbon dots

    SciTech Connect

    Liu, Ruihua; Li, Haitao; Kong, Weiqian; Liu, Juan; Liu, Yang; Tong, Cuiyan; Zhang, Xing; Kang, Zhenhui

    2013-07-15

    Graphical abstract: Fluorescent carbon dots were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from PEG and demonstrated to show high selectivity toward Hg2+ ions detection. - Highlights: • FCDs were synthesized by one-step sodium hydroxide-assisted reflux method from PEG. • The FCDs emit blue photoluminescence and have upconversion fluorescent property. • The FCDs show ultra-sensitive detective ability for Hg{sup 2+} ions. - Abstract: Fluorescent carbon dots (FCDs) were efficiently synthesized by one-step sodium hydroxide-assisted reflux method from poly(ethylene glycol) (PEG). The obtained FCDs exhibit excellent water-solubility and high stability. Under the UV irradiation, the FCDs could emit bright blue photoluminescence, and also they were found to show excellent up-conversion fluorescence. It was further demonstrated that such FCDs can serve as effective fluorescent sensing platform for Hg{sup 2+} ions detection with ultra-sensitivity and selectivity. The sensing system achieved a limit of detection as low as 1 fM, which is much lower than all the previous reported sensing systems for Hg{sup 2+} ions detection. This FCDs sensing system has been successfully applied for the analysis of Hg{sup 2+} ions in water samples from river, lake, and tap water, showing good practical feasibility.

  1. Thermal Outlining using Focused Ultrasound (TOFU) with reversible temperature sensitive fluorescent probes

    NASA Astrophysics Data System (ADS)

    Kwong, Tiffany C.; Nouizi, Farouk; Lin, Yuting; Zhu, Yue; Sampathkumaran, Uma; Gulsen, Gultekin

    2016-03-01

    Optical imaging has long been hindered by the high absorption and scattering of light in biological tissue. This makes it difficult to probe beyond a few millimeters beneath the surface without sacrificing image resolution and quantitative accuracy. Strong scattering and the inherent nature of the inverse problem makes fluorescence diffuse optical tomography (FT) extremely challenging. To this end, multi-modality techniques that combine anatomical imaging with the functional optical information have been used to improve the resolution and accuracy of FT. Previously, we have reported on the feasibility of a new imaging method, "Thermal Outlining using Focused Ultrasound" (TOFU), which combines the sensitivity of FT with the resolution of focused ultrasound using temperature reversible fluorescent probes. In this method, the position of the temperature reversible fluorescent probes is localized by an increase in fluorescent signal when the hot spot of the focused ultrasound beam is scanned over the medium. This a priori information is then utilized to guide and constrain conventional reconstruction algorithm to recover the position and concentration of the probes more accurately. The small size of the focal spot (~1.4 mm) up to a depth of 6 cm, allows imaging the distribution of these temperature sensitive agents with not only high spatial resolution but also high quantitative accuracy in deep tissue. In this work, the performance of the system will be evaluated using simulation and phantoms to investigate the dependence that size of the fluorescent distribution has on the TOFU system performance.

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

    PubMed

    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.

  3. Mesoporous structured MIPs@CDs fluorescence sensor for highly sensitive detection of TNT.

    PubMed

    Xu, Shoufang; Lu, Hongzhi

    2016-11-15

    A facile strategy was developed to prepare mesoporous structured molecularly imprinted polymers capped carbon dots (M-MIPs@CDs) fluorescence sensor for highly sensitive and selective determination of TNT. The strategy using amino-CDs directly as "functional monomer" for imprinting simplify the imprinting process and provide well recognition sites accessibility. The as-prepared M-MIPs@CDs sensor, using periodic mesoporous silica as imprinting matrix, and amino-CDs directly as "functional monomer", exhibited excellent selectivity and sensitivity toward TNT with detection limit of 17nM. The recycling process was sustainable for 10 times without obvious efficiency decrease. The feasibility of the developed method in real samples was successfully evaluated through the analysis of TNT in soil and water samples with satisfactory recoveries of 88.6-95.7%. The method proposed in this work was proved to be a convenient and practical way to prepare high sensitive and selective fluorescence MIPs@CDs sensors.

  4. Rapid and sensitive determination of clenbuterol in porcine muscle and swine urine using a fluorescent probe

    NASA Astrophysics Data System (ADS)

    Jing, Xu; Bai, Bing; Zhang, Chenxuan; Wu, Wenying; Du, Liming; Liu, Hailong; Yao, Guojun

    2015-02-01

    The feed additive Clenbuterol hydrochloric acid (CLB) is non-fluorescent, thus it is difficult to quantify through direct fluorescent method. Palmatine (PAL) can react with cucurbit[7]uril (CB[7]) to form stable complexes as a fluorescent probe. Significant quenching of the fluorescence intensity of the CB[7]-PAL complex was observed with the addition of CLB. Based on the significant quenching of the supramolecular complex fluorescence intensity, a novel spectrofluorimetric method with high convenience, selectivity and sensitivity was developed for the determination of CLB. The fluorescence quenching values (ΔF) showed good linear relationship with CLB concentrations from 0.011 μg mL-1 to 4.2 μg mL-1 with a detection limit 0.004 μg mL-1. In this research, an ultrasound treatment replaced the former time-consuming shake method to form stable complexes. The proposed spectrofluorimetric method had been successfully applied to the determination of CLB in porcine muscle and swine urine with good precision and accuracy. The competing reaction and the supramolecular interaction mechanisms between the CLB and PAL as they fight for occupancy of the CB[7] cavity were studied using spectrofluorimetry, 1H NMR, and molecular modeling calculations. Interestingly, results indicate that two stable CB[7]-CLB complexes were formed.

  5. Rapid and sensitive determination of clenbuterol in porcine muscle and swine urine using a fluorescent probe.

    PubMed

    Jing, Xu; Bai, Bing; Zhang, Chenxuan; Wu, Wenying; Du, Liming; Liu, Hailong; Yao, Guojun

    2015-02-05

    The feed additive Clenbuterol hydrochloric acid (CLB) is non-fluorescent, thus it is difficult to quantify through direct fluorescent method. Palmatine (PAL) can react with cucurbit[7]uril (CB[7]) to form stable complexes as a fluorescent probe. Significant quenching of the fluorescence intensity of the CB[7]-PAL complex was observed with the addition of CLB. Based on the significant quenching of the supramolecular complex fluorescence intensity, a novel spectrofluorimetric method with high convenience, selectivity and sensitivity was developed for the determination of CLB. The fluorescence quenching values (ΔF) showed good linear relationship with CLB concentrations from 0.011 μg mL(-1) to 4.2 μg mL(-1) with a detection limit 0.004 μg mL(-1). In this research, an ultrasound treatment replaced the former time-consuming shake method to form stable complexes. The proposed spectrofluorimetric method had been successfully applied to the determination of CLB in porcine muscle and swine urine with good precision and accuracy. The competing reaction and the supramolecular interaction mechanisms between the CLB and PAL as they fight for occupancy of the CB[7] cavity were studied using spectrofluorimetry, (1)H NMR, and molecular modeling calculations. Interestingly, results indicate that two stable CB[7]-CLB complexes were formed.

  6. A turn-on fluorescent indicator for citrate with micromolar sensitivity.

    PubMed

    Burguete, M Isabel; Galindo, Francisco; Luis, Santiago V; Vigara, Laura

    2007-09-28

    A turn-on fluorescent indicator for citric acid (citrate) has been developed, displaying high emission enhancement (+1500%) and low interference by other carboxylates. The sensor is based on the non-emissive copper(II) complex of a fluorescent amino amide, which, upon addition of citrate decomplexates to yield the emissive ligand. The detection limit estimated for this new chemosensing system is about 0.5 microM. This novel approach to the analysis of citrate constitutes an alternative ca. 10(2)-10(3) times more sensitive than the standard method based on the enzyme citrate lyase.

  7. A Novel Fluorescent Probe for the Highly Selective and Sensitive Detection of Palladium in Aqueous Medium.

    PubMed

    Ma, Zhiwei; Wang, Xiao; Sun, Yanling; Liu, Juntao; Tong, Yan; Liu, Zhijing

    2016-11-01

    Based on the Pd(0)-catalyzed Tsuji-Trost allylic oxidative insertion reaction, we developed a fluorescent probe PdL1 for sensing Pd(0). As expected, probe PdL1 exhibited high selectivity and excellent sensitivity in both absorbance and fluorescence detection of Pd(0) in CH3CH2OH/PBS (10 mM, pH = 7.4, 6:4, v/v) solution. The detection limit was calculated to be as low as 15 nM, which can meet the selective requirements for practical application.

  8. Design considerations for an x-ray microprobe

    SciTech Connect

    Howells, M.R.; Hastings, J.B.

    1982-01-01

    The optical design of a fluorescent microprobe covering the x-ray region from 2 to 16 keV is considered for the NSLS x-ray ring. The limit on detectability is from total flux (photons/..mu..m/sup 2/) and several design choices are considered to match the optical system to the storage ring to maximize throughput. The tradeoffs in image quality and energy resolution of these designs have been considered and within these constraints two firm proposals are presented.

  9. An x-ray microprobe beam line for trace element analysis

    SciTech Connect

    Gordon, B.M.; Hanson, A.L.; Jones, K.W.; Kwiatek, W.M.; Long, G.J.; Pounds, J.G.; Schidlovsky, G.; Spanne, P.; Rivers, M.L.; Sutton, S.R.

    1987-01-01

    The application of synchrotron radiation to an x-ray microprobe for trace element analysis is a complementary and natural extension of existing microprobe techniques using electrons, protons, and heavier ions as excitation sources for x-ray fluorescence. The ability to focus charged particles leads to electron microprobes with spatial resolutions in the sub-micrometer range and down to 100 ppM detection limits and proton microprobes with micrometer resolution and ppM detection limits. The characteristics of synchrotron radiation that prove useful for microprobe analysis include a broad and continuous energy spectrum, a relatively small amount of radiation damage compared to that deposited by charged particles, a highly polarized source which reduces background scattered radiation in an appropriate counting geometry, and a small vertical divergence angle of approx.0.2 mrad which allows for focussing of the light beam into a small spot with high flux. The features of a dedicated x-ray microprobe beam line developed at the National Synchrotron Light Source (NSLS) are described. 4 refs., 3 figs.

  10. A 4-methylumbelliferone-based fluorescent probe for the sensitive detection of captopril.

    PubMed

    Wang, Liping; Yang, Xiao-Feng; Zhao, Minglei

    2009-07-01

    A highly sensitive fluorogenic probe for captopril, 4-methylumbelliferyl-2, 4-dinitrobenzenesulfonate (4-MUDNBS), was designed and synthesized. 4-MUDNBS is a nonfluorescent compound and was synthesized via the one-step reaction of 4-methylumbelliferone (4-MU) with 2,4-dinitrobenzenesulfonyl chloride. Upon mixing with captopril in basic solution, the 2,4-dinitrobenzenesulfonyl group of 4-MUDNBS was efficiently removed and highly fluorescent 4-MU was released, hence leading to the dramatic fluorescence increase of the reaction solution. The fluorescence intensity is linear with captopril concentration in the range 3.0-500 ng mL(-1) with a detection limit of 2.2 ng mL(-1) (3sigma). The effect of substituents on the benzenesulfonyl moiety of the probe is discussed, and the presence of electronegative groups is favorable for the thiolate-induced cleavage reaction. The proposed method has been successfully applied to the captopril determination in pharmaceutical preparations.

  11. Detection of coalescing agents in water-borne latex emulsions using an environment sensitive fluorescent probe.

    PubMed

    Raja, Tanzeela Nazir; Brouwer, Albert M; Biemans, Koen; Nabuurs, Tijs; Tennebroek, Ronald

    2010-07-30

    In this paper we report the determination of partitioning of coalescing agents (organic co-solvents) in water-borne latex emulsions by means of a fluorescence method. An environment-sensitive fluorescent probe was copolymerized via emulsion polymerization. The presence of organic co-solvents inside the polymer particles is revealed by the photophysical properties of the probe. In particular, the position of the fluorescence emission maximum of co-polymerized can be used to measure the amount of coalescing agent present in the polymer particles. The spectral shifts are shown to be due to the softening of the matrix, rather than to solvation of the probe by the added co-solvent.

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

    NASA Astrophysics Data System (ADS)

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

    2014-12-01

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

  13. Charge-Transfer Dynamics of Fluorescent Dye-Sensitized Electrodes under Applied Biases.

    PubMed

    Godin, Robert; Sherman, Benjamin D; Bergkamp, Jesse J; Chesta, Carlos A; Moore, Ana L; Moore, Thomas A; Palacios, Rodrigo E; Cosa, Gonzalo

    2015-07-16

    The development of dye-sensitized solar cells requires an in-depth understanding of the interfacial charge-transfer dynamics that take place between dye sensitizers and semiconductors. Here, we describe a prototype system to probe these dynamics by monitoring in real time the fluorescence of two organic sensitizers, a perylene and a squaraine, bound to a SnO2 semiconductor thin film as a function of potentiostatic control of the Fermi level. The two different sensitizer fluorophores characterized by vastly different redox potentials undergo similar fluorescence modulation with applied bias, an indication that the density of states of the semiconductor largely influences the charge-transfer dynamics while energetics play a minimal role. We further show that the rate of photodegradation of the perylene sensitizer with applied bias provides a suitable marker to study the rate of charge injection and charge recombination. Taken together, our results demonstrate a suitable platform to visualize and study charge-transfer dynamics on films and constitute a step toward achieving single-molecule resolution in our quest to decipher the static and dynamic heterogeneity of charge-transfer dynamics in dye-sensitized photoanodes.

  14. New environment-sensitive multichannel DNA fluorescent label for investigation of the protein-DNA interactions.

    PubMed

    Kuznetsova, Alexandra A; Kuznetsov, Nikita A; Vorobjev, Yuri N; Barthes, Nicolas P F; Michel, Benoît Y; Burger, Alain; Fedorova, Olga S

    2014-01-01

    Here, we report the study of a new multichannel DNA fluorescent base analogue 3-hydroxychromone (3HC) to evaluate its suitability as a fluorescent reporter probe of structural transitions during protein-DNA interactions and its comparison with the current commercially available 2-aminopurine (aPu), pyrrolocytosine (Cpy) and 1,3-diaza-2-oxophenoxazine (tCO). For this purpose, fluorescent base analogues were incorporated into DNA helix on the opposite or on the 5'-side of the damaged nucleoside 5,6-dihydrouridine (DHU), which is specifically recognized and removed by Endonuclease VIII. These fluorophores demonstrated different sensitivities to the DNA helix conformational changes. The highest sensitivity and the most detailed information about the conformational changes of DNA induced by protein binding and processing were obtained using the 3HC probe. The application of this new artificial fluorescent DNA base is a very useful tool for the studies of complex mechanisms of protein-DNA interactions. Using 3HC biosensor, the kinetic mechanism of Endonuclease VIII action was specified.

  15. Determination of L-tyrosine by β-cyclodextrin sensitized fluorescence quenching method

    NASA Astrophysics Data System (ADS)

    Zhu, Xiashi; Xu, Suqin

    2010-10-01

    A novel β-cyclodextrin (β-CD) sensitized fluorescence quenching method for the determination of L-tyrosine ( L-Tyr) with Mo(VI)-phenyl-fluorone (PF) as a fluorescence probe has been developed. The fluorescence intensity of Mo(VI)-PF-β-CD was diminished as the L-tyrosine was added, the fluorescence quenching value Δ F = Fβ-CD-Mo-PF - Fβ-CD-Mo-PF- L-Tyr was enhanced in β-CD and there was a linear relationship between the Δ F and the concentration of L-Tyr. Under the optimal conditions, the linear range of calibration curve for the determination of L-tyrosine was 0.3-20.0 μg mL -1; the detection limit was 0.094 μg mL -1. NaOH (10%, w/v) is the best reagent of hydrolysis in sample preparation. The sensitized mechanism of β-cyclodextrin was discussed. The method has been applied to the determination of L-tyrosine in spirulina and food samples with satisfactory results.

  16. Overlap of Doxycycline Fluorescence with that of the Redox-Sensitive Intracellular Reporter roGFP.

    PubMed

    Khader, Heba; Solodushko, Victor; Al-Mehdi, Abu Bakr; Audia, Jonathon; Fouty, Brian

    2014-03-01

    Tetracycline-inducible systems allow for either suppression or induction of transgene expression to facilitate studies of cell physiology. Doxycycline is a preferred inducer for these gene expression systems due to its membrane permeability; however, the heterocyclic structure of doxycycline exhibits fluorogenic properties that can potentially bias measurement of other fluorochromes. Thus the simultaneous use of tetracycline-inducible systems and fluorescent proteins as reporter genes or as intracellular biosensors may lead to potentially confounding results. Herein, using cells which co-express the ratiometric redox sensitive intracellular reporter, roGFP, and a tetracycline-inducible reporter plasmid encoding the reporter gene, mCherry, as a model system, we describe the overlapping intracellular fluorescent signals between doxycycline and commonly used intracellular fluorescent probes. In our cells, the addition of doxycycline to cells caused a dose- and time-dependent increase in cell fluorescence with 405 nm excitation which overlapped with that of the oxidized configuration of roGFP. Incubating cells in concentrations of doxycycline less than 1 μg/mL and removing doxycycline from the media 60 min before performing experiments eliminated fluorescence interference while still maintaining maximal reporter transgene activation.

  17. Polythiophene nanofilms for sensitive fluorescence detection of viruses in drinking water.

    PubMed

    Wankar, Shashwati; Turner, Nicholas W; Krupadam, Reddithota J

    2016-08-15

    Molecular imprints of the tobacco necrosis virus (TNV) have been formed within polythiophene nanofilms with an approximate thickness of 200nm. These films have been electrochemically deposited onto conducting Au surfaces. Upon rebinding, the TNV-polythiophene complex changes the fluorescence intensity of the nanofilm. The fluorescence intensity at 410nm was observed to be proportional to the concentration of viruses in the range of 0.1-10ngL(-1) (0.15-15pg) with the lower calculated detection limit of 2.29ngL(-1) (3.4pg). The intensity of the fluorescence emission is not affected by the thickness of the polythiophene film and the nature of TNV specific binding sites. Kinetic data analyses showed that the nanofilm responds to TNV within 2min; and cross-selectivity studies with tobacco mosaic virus (TMV) showed an excellent specificity for the targeted TNV. These binding experiments demonstrate the potential of fluorescence emission for the specific, label free and rapid detection of viruses using nanofilm sensors. Taking into account the lower limit of detection, the fluorescence sensing reported here is reliable, simple to perform, rapid, cost-effective and offers a sensitive analytical method for virus detection in water resources.

  18. Luminescent conjugated oligothiophenes for sensitive fluorescent assignment of protein inclusion bodies.

    PubMed

    Klingstedt, Therése; Blechschmidt, Cristiane; Nogalska, Anna; Prokop, Stefan; Häggqvist, Bo; Danielsson, Olof; Engel, W King; Askanas, Valerie; Heppner, Frank L; Nilsson, K Peter R

    2013-03-18

    Small hydrophobic ligands identifying intracellular protein deposits are of great interest, as protein inclusion bodies are the pathological hallmark of several degenerative diseases. Here we report that fluorescent amyloid ligands, termed luminescent conjugated oligothiophenes (LCOs), rapidly and with high sensitivity detect protein inclusion bodies in skeletal muscle tissue from patients with sporadic inclusion body myositis (s-IBM). LCOs having a conjugated backbone of at least five thiophene units emitted strong fluorescence upon binding, and showed co-localization with proteins reported to accumulate in s-IBM protein inclusion bodies. Compared with conventional amyloid ligands, LCOs identified a larger fraction of immunopositive inclusion bodies. When the conjugated thiophene backbone was extended with terminal carboxyl groups, the LCO revealed striking spectral differences between distinct protein inclusion bodies. We conclude that 1) LCOs are sensitive, rapid and powerful tools for identifying protein inclusion bodies and 2) LCOs identify a wider range of protein inclusion bodies than conventional amyloid ligands.

  19. Terbium-sensitized fluorescence method for the determination of deferasirox in biological fluids and tablet formulation.

    PubMed

    Manzoori, Jamshid L; Jouyban, Abolghasem; Amjadi, Mohammad; Panahi-Azar, Vahid; Tamizi, Elnaz; Vaez-Gharamaleki, Jalil

    2011-01-01

    A novel, rapid and sensitive spectroflurimetric method was developed and validated for the determination of deferasirox in urine, serum and tablet samples based on sensitization of terbium fluorescence. The excitation and emission wavelengths were 328 and 545 nm, respectively. The optimum conditions for the determination of deferasirox were investigated considering the effects of various parameters. The method was quantitatively evaluated in terms of linearity, recovery, reproducibility and limit of detection. Under the optimal conditions, the fluorescence intensities were linear with the concentration of deferasirox in the range of 5 × 10(-9) to 5×10(-6) mol L(-1) , with a detection limit of 1.5 × 10(-9) mol L(-1) and a relative standard deviation of 1.1-2.3%. Linearity, reproducibility, recovery and limit of detection made the method suitable for determination of deferasirox in urine, serum and tablets samples.

  20. Improved Diffuse Fluorescence Flow Cytometer Prototype for High Sensitivity Detection of Rare Circulating Cells In Vivo

    NASA Astrophysics Data System (ADS)

    Pestana, Noah Benjamin

    Accurate quantification of circulating cell populations is important in many areas of pre-clinical and clinical biomedical research, for example, in the study of cancer metastasis or the immune response following tissue and organ transplants. Normally this is done "ex-vivo" by drawing and purifying a small volume of blood and then analyzing it with flow cytometry, hemocytometry or microfludic devices, but the sensitivity of these techniques are poor and the process of handling samples has been shown to affect cell viability and behavior. More recently "in vivo flow cytometry" (IVFC) techniques have been developed where fluorescently-labeled cells flowing in a small blood vessel in the ear or retina are analyzed, but the sensitivity is generally poor due to the small sampling volume. To address this, our group recently developed a method known as "Diffuse Fluorescence Flow Cytometry" (DFFC) that allows detection and counting of rare circulating cells with diffuse photons, offering extremely high single cell counting sensitivity. In this thesis, an improved DFFC prototype was designed and validated. The chief improvements were three-fold, i) improved optical collection efficiency, ii) improved detection electronics, and iii) development of a method to mitigate motion artifacts during in vivo measurements. In combination, these improvements yielded an overall instrument detection sensitivity better than 1 cell/mL in vivo, which is the most sensitive IVFC system reported to date. Second, development and validation of a low-cost microfluidic device reader for analysis of ocular fluids is described. We demonstrate that this device has equivalent or better sensitivity and accuracy compared a fluorescence microscope, but at an order-of-magnitude reduced cost with simplified operation. Future improvements to both instruments are also discussed.

  1. Fluorescent Immunochromatography for Rapid and Sensitive Typing of Seasonal Influenza Viruses

    PubMed Central

    Sakurai, Akira; Takayama, Katsuyoshi; Nomura, Namiko; Kajiwara, Naoki; Okamatsu, Masatoshi; Yamamoto, Naoki; Tamura, Tsuruki; Yamada, Jitsuho; Hashimoto, Masako; Sakoda, Yoshihiro; Suda, Yoshihiko; Kobayashi, Yukuharu; Kida, Hiroshi; Shibasaki, Futoshi

    2015-01-01

    Lateral flow tests also known as Immunochromatography (IC) is an antigen-detection method conducted on a nitrocellulose membrane that can be completed in less than 20 min. IC has been used as an important rapid test for clinical diagnosis and surveillance of influenza viruses, but the IC sensitivity is relatively low (approximately 60%) and the limit of detection (LOD) is as low as 10³ pfu per reaction. Recently, we reported an improved IC assay using antibodies conjugated with fluorescent beads (fluorescent immunochromatography; FLIC) for subtyping H5 influenza viruses (FLIC-H5). Although the FLIC strip must be scanned using a fluorescent reader, the sensitivity (LOD) is significantly improved over that of conventional IC methods. In addition, the antibodies which are specific against the subtypes of influenza viruses cannot be available for the detection of other subtypes when the major antigenicity will be changed. In this study, we established the use of FLIC to type seasonal influenza A and B viruses (FLIC-AB). This method has improved sensitivity to 100-fold higher than that of conventional IC methods when we used several strains of influenza viruses. In addition, FLIC-AB demonstrated the ability to detect influenza type A and influenza type B viruses from clinical samples with high sensitivity and specificity (Type A: sensitivity 98.7% (74/75), specificity 100% (54/54), Type B: sensitivity 100% (90/90), specificity 98.2% (54/55) in nasal swab samples) in comparison to the results of qRT-PCR. And furthermore, FLIC-AB performs better in the detection of early stage infection (under 13h) than other conventional IC methods. Our results provide new strategies to prevent the early-stage transmission of influenza viruses in humans during both seasonal outbreaks and pandemics. PMID:25650570

  2. Highly sensitive ratiometric fluorescent chemosensor for silver ion and silver nanoparticles in aqueous solution.

    PubMed

    Jang, Sujung; Thirupathi, Ponnaboina; Neupane, Lok Nath; Seong, Junho; Lee, Hyunsook; Lee, Wan In; Lee, Keun-Hyeung

    2012-09-21

    A pyrene derivative chemosensor (Pyr-WH) based on a dipeptide shows a highly sensitive ratiometric response to Ag(I) as well as silver nanoparticles in aqueous solution at physiological pH. Pyr-WH penetrated live HeLa cells and exhibits a ratiometric response to intracellular Ag(I). The binding mode of Pyr-WH with Ag(I) was characterized based on fluorescence changes in different pH, NMR, and ESI mass spectrometer experiments.

  3. On the analysis of neonatal hamster tooth germs with the photon microprobe at Daresbury, UK

    NASA Astrophysics Data System (ADS)

    Tros, G. H. J.; Van Langevelde, F.; Vis, R. D.

    1990-04-01

    Complementary to the micro-PIXE experiments performed on hamster tooth germs to elucidate the role of fluoride during the growth, the photon microprobe at Daresbury was used to obtain information on the distribution of Zn. The germs of fluoride-administered hamsters, together with a control group, were analyzed with the micro-synchrotron radiation fluorescence method (micro-SXRF).

  4. TEAM - Titan Exploration Atmospheric Microprobes

    NASA Astrophysics Data System (ADS)

    Nixon, Conor; Esper, Jaime; Aslam, Shahid; Quilligan, Gerald

    2016-10-01

    The astrobiological potential of Titan's surface hydrocarbon liquids and probable interior water ocean has led to its inclusion as a destination in NASA's "Ocean Worlds" initiative, and near-term investigation of these regions is a high-level scientific goal. TEAM is a novel initiative to investigate the lake and sea environs using multiple dropsondes -scientific probes derived from an existing cubesat bus architecture (CAPE - the Cubesat Application for Planetary Exploration) developed at NASA GSFC. Each 3U probe will parachute to the surface, making atmospheric structure and composition measurements during the descent, and photographing the surface - land, shoreline and seas - in detail. TEAM probes offer a low-cost, high-return means to explore multiple areas on Titan, yielding crucial data about the condensing chemicals, haze and cloud layers, winds, and surface features of the lakes and seas. These microprobes may be included on a near-term New Frontiers class mission to the Saturn system as additional payload, bringing increased scientific return and conducting reconnaissance for future landing zones. In this presentation we describe the probe architecture, baseline payload, flight profile and the unique engineering and science data that can be returned.

  5. A viscosity sensitive fluorescent dye for real-time monitoring of mitochondria transport in neurons.

    PubMed

    Baek, Yeonju; Park, Sang Jun; Zhou, Xin; Kim, Gyungmi; Kim, Hwan Myung; Yoon, Juyoung

    2016-12-15

    We present here a viscosity sensitive fluorescent dye, namely thiophene dihemicyanine (TDHC), that enables the specific staining of mitochondria. In comparison to the common mitochondria tracker (Mitotracker Deep Red, MTDR), this dye demonstrated its unique ability for robust staining of mitochondria with high photostability and ultrahigh signal-to-noise ratio (SNR). Moreover, TDHC also showed high sensitivity towards mitochondria membrane potential (ΔΨm) and intramitochondria viscosity change. Consequently, this dye was utilized in real-time monitoring of mitochondria transport in primary cortical neurons. Finally, the Two-Photon Microscopy (TPM) imaging ability of TDHC was also demonstrated.

  6. Highly adaptable and sensitive protease assay based on fluorescence resonance energy transfer.

    PubMed

    Zauner, Thomas; Berger-Hoffmann, Renate; Müller, Katrin; Hoffmann, Ralf; Zuchner, Thole

    2011-10-01

    Proteases are widely used in analytical sciences and play a central role in several widespread diseases. Thus, there is an immense need for highly adaptable and sensitive assays for the detection and monitoring of various proteolytic enzymes. We established a simple protease fluorescence resonance energy transfer (pro-FRET) assay for the determination of protease activities, which could in principle be adapted for the detection of all proteases. As proof of principle, we demonstrated the potential of our method using trypsin and enteropeptidase in complex biological mixtures. Briefly, the assay is based on the cleavage of a FRET peptide substrate, which results in a dramatic increase of the donor fluorescence. The assay was highly sensitive and fast for both proteases. The detection limits for trypsin and enteropeptidase in Escherichia coli lysate were 100 and 10 amol, respectively. The improved sensitivity for enteropeptidase was due to the application of an enzyme cascade, which leads to signal amplification. The pro-FRET assay is highly specific as even high concentrations of other proteases did not result in significant background signals. In conclusion, this sensitive and simple assay can be performed in complex biological mixtures and can be easily adapted to act as a versatile tool for the sensitive detection of proteases.

  7. Solid state photon upconversion utilizing thermally activated delayed fluorescence molecules as triplet sensitizer

    SciTech Connect

    Wu, Tony C.; Congreve, Daniel N.; Baldo, Marc A.

    2015-07-20

    The ability to upconvert light is useful for a range of applications, from biological imaging to solar cells. But modern technologies have struggled to upconvert incoherent incident light at low intensities. Here, we report solid state photon upconversion employing triplet-triplet exciton annihilation in an organic semiconductor, sensitized by a thermally activated-delayed fluorescence (TADF) dye. Compared to conventional phosphorescent sensitizers, the TADF dye maximizes the wavelength shift in upconversion due to its small singlet-triplet splitting. The efficiency of energy transfer from the TADF dye is 9.1%, and the conversion yield of sensitizer exciton pairs to singlet excitons in the annihilator is 1.1%. Our results demonstrate upconversion in solid state geometries and with non-heavy metal-based sensitizer materials.

  8. The ANSTO high energy heavy ion microprobe

    NASA Astrophysics Data System (ADS)

    Siegele, Rainer; Cohen, David D.; Dytlewski, Nick

    1999-10-01

    Recently the construction of the ANSTO High Energy Heavy Ion Microprobe (HIMP) at the 10 MV ANTARES tandem accelerator has been completed. The high energy heavy ion microprobe focuses not only light ions at energies of 2-3 MeV, but is also capable of focusing heavy ions at high energies with ME/ q2 values up to 150 MeV amu and greater. First performance tests and results are reported here.

  9. Fluorescence Based Characterization of Calcium Sensitizer Action on the Troponin Complex.

    PubMed

    Schlecht, William; Li, King-Lun; Hu, Dehong; Dong, Wenji

    2016-02-01

    Calcium sensitizers enhance the transduction of the Ca(2+) signal into force within the heart and have found use in treating heart failure. However the mechanisms of action for most Ca(2+) sensitizers remain unclear. To address this issue an efficient fluorescence based approach to Ca(2+) sensitizer screening was developed which monitors cardiac troponin C's (cTnC's) hydrophobic cleft. This approach was tested on four common Ca(2+) -sensitizers, EMD 57033, levosimendan, bepridil and pimobendan with the aim of elucidating the mechanisms of action for each as well as proving the efficacy of the new screening method. Ca(2+) -titration experiments were employed to determine the effect on Ca(2+) sensitivity and cooperativity of cTnC opening, while stopped flow experiments were used to investigate the impact on cTnC relaxation kinetics. Bepridil was shown to increase the sensitivity of cTnC for Ca(2+) under all reconstitution conditions, sensitization by the other drugs was context dependent. Levosimendan and pimobendan reduced the rate of cTnC closing consistent with a stabilization of cTnC's open conformation while bepridil increased the rate of relaxation. Experiments were also run on samples containing cTnT(T204E), a known Ca(2+) -desensitizing phosphorylation mimic. Levosimendan, bepridil, and pimobendan were found to elevate the Ca(2+) -sensitivity of cTnT(T204E) containing samples in this context.

  10. Highly sensitive detection of human papillomavirus type 16 DNA using time-resolved fluorescence microscopy and long lifetime probes

    NASA Astrophysics Data System (ADS)

    Wang, Xue F.; Periasamy, Ammasi; Wodnicki, Pawel; Siadat-Pajouh, M.; Herman, Brian

    1995-04-01

    We have been interested in the role of Human Papillomavirus (HPV) in cervical cancer and its diagnosis; to that end we have been developing microscopic imaging and fluorescent in situ hybridization (FISH) techniques to genotype and quantitate the amount of HPV present at a single cell level in cervical PAP smears. However, we have found that low levels of HPV DNA are difficult to detect accurately because theoretically obtainable sensitivity is never achieved due to nonspecific autofluorescence, fixative induced fluorescence of cells and tissues, and autofluorescence of the optical components in the microscopic system. In addition, the absorption stains used for PAP smears are intensely autofluorescent. Autofluorescence is a rapidly decaying process with lifetimes in the range of 1-100 nsec, whereas phosphorescence and delayed fluorescence have lifetimes in the range of 1 microsecond(s) ec-10 msec. The ability to discriminate between specific fluorescence and autofluorescence in the time-domain has improved the sensitivity of diagnostic test such that they perform comparably to, or even more sensitive than radioisotopic assays. We have developed a novel time-resolved fluorescence microscope to improve the sensitivity of detection of specific molecules of interest in slide based specimens. This time-resolved fluorescence microscope is based on our recently developed fluorescence lifetime imaging microscopy (FILM) in conjunction with the use of long lifetime fluorescent labels. By using fluorescence in situ hybridization and the long lifetime probe (europium), we have demonstrated the utility of this technique for detection of HPV DNA in cervicovaginal cells. Our results indicate that the use of time-resolved fluorescence microscopy and long lifetime probes increases the sensitivity of detection by removing autofluorescence and will thus lead to improved early diagnosis of cervical cancer. Since the highly sensitive detection of DNA in clinical samples using

  11. Assessment of alginate hydrogel degradation in biological tissue using viscosity-sensitive fluorescent dyes

    NASA Astrophysics Data System (ADS)

    Shkand, Tatiana V.; Chizh, Mykola O.; Sleta, Iryna V.; Sandomirsky, Borys P.; Tatarets, Anatoliy L.; Patsenker, Leonid D.

    2016-12-01

    The main goal of this study is to investigate a combination of viscosity-sensitive and viscosity-insensitive fluorescent dyes to distinguish different rheological states of hydrogel based biostructural materials and carriers in biological tissues and to assess their corresponding location areas. The research is done in the example of alginate hydrogel stained with viscosity-sensitive dyes Seta-470 and Seta-560 as well as the viscosity-insensitive dye Seta-650. These dyes absorb/emit at 469/518, 565/591 and 651/670 nm, respectively. The rheological state of the alginate, the area of the fluorescence signal and the mass of the dense alginate versus the calcium gluconate concentration utilized for alginate gelation were studied in vitro. The most pronounced change in the fluorescence signal area was found at the same concentrations of calcium gluconate (below ~1%) as the change in the alginate plaque mass. The stained alginate was also implanted in situ in rat hip and myocardium and monitored using fluorescence imaging. In summary, our data indicate that the viscosity sensitive dye in combination with the viscosity-insensitive dye allow tracking the biodegradation of the alginate hydrogel and determining the rheological state of hydrogel in biological tissue, which both should have relevance for research and clinical applications. Using this method we estimated the half-life of the dense alginate hydrogel in a rat hip to be in the order of 4 d and about 6-8 d in rat myocardium. The half-life of the dense hydrogel in the myocardium was found to be long enough to prevent aneurysm rupture of the left ventricle wall, one of the more severe complications of the early post-infarction period.

  12. pH sensitive CdS-iron oxide fluorescent-magnetic nanocomposites.

    PubMed

    Thakur, Dhananjay; Deng, Shuang; Baldet, Thierno; Winter, Jessica O

    2009-12-02

    There has been great interest in the use of nanoparticles for imaging, particularly in multimodal applications (e.g., combination of MRI and fluorescence). Yet creating particles with multiple functionalities has been challenging. Here, we report the synthesis of pH sensitive, fluorescent-magnetic, nanocomposites created through a simple aqueous procedure. Separately synthesized superparamagnetic iron oxide nanoparticles and mercaptopropionic acid (MPA)-coated CdS quantum dots were crosslinked using 3-mercaptopropyl trimethoxysilane (MPS) as a bifunctional linker to yield CdS-iron oxide conjugates. Conjugates formed clusters of 0.1-1.0 microm diameter, with the smallest observed particle diameter approximately 50 nm. Particle solubility and photoluminescent (PL) intensity were sensitive to solution pH, with the highest PL intensity and stability obtained at pH values < 3.0 and MPS:Cd:Fe ratios of 1:10:1. pH sensitivity is believed to result from changes in nanoparticle solubility within the silica-based matrix. Given these unique properties, this material might find application in separation, pH sensitive detection (e.g., endosomal tracking) and biosensing.

  13. Discovery of a Novel Fluorescent Probe for the Sensitive Detection of β-Amyloid Deposits

    PubMed Central

    Ren, Wenming; Xu, Mingming; Liang, Steven H.; Xiang, Huaijiang; Tang, Li; Zhang, Minkui; Ding, Dejun; Li, Xin; Zhang, Haiyan; Hu, Youhong

    2016-01-01

    Here we reported the development of the first photoinduced electron transfer (PeT) probe (1) to directly locate β-amyloid aggregates (Aβ plaques) in the brain without the need of post-washing procedures. The probe showed a high affinity for Aβ aggregates with a Kd value of 3.5 nM. It is weakly emissive by itself with its fluorescence quenched by electron transfer from PeT donor to the excited fluorophore. But selective binding to Aβ plaques would attenuate the PeT process and restore the fluorescence, therefore facilitating the tracking of Aβ plaques. The probe is advantageous in that its fluorescence is environment-less-sensitive and no washing procedure is required to provide high contrast fluorescent signal when applied to stain brain tissues. As a proof of concept, its application has been exemplified by staining Aβ plaques in slices of brain tissue from double transgenic (APP/PS1) mice of Alzheimer’s disease. PMID:26313423

  14. Quantification of nanoparticle endocytosis based on double fluorescent pH-sensitive nanoparticles.

    PubMed

    Kurtz-Chalot, Andréa; Klein, Jean-Philippe; Pourchez, Jérémie; Boudard, Delphine; Bin, Valérie; Sabido, Odile; Marmuse, Laurence; Cottier, Michèle; Forest, Valérie

    2015-04-01

    Amorphous silica is a particularly interesting material because of its inertness and chemical stability. Silica nanoparticles have been recently developed for biomedical purposes but their innocuousness must be carefully investigated before clinical use. The relationship between nanoparticles physicochemical features, their uptake by cells and their biological activity represents a crucial issue, especially for the development of nanomedicine. This work aimed at adapting a method for the quantification of nanoparticle endocytosis based on pH-sensitive and double fluorescent particles. For that purpose, silica nanoparticles containing two fluorophores: FITC and pHrodo(TM) were developed, their respective fluorescence emission depends on the external pH. Indeed, FITC emits a green fluorescence at physiological pH and pHrodo(TM) emits a red fluorescence which intensity increased with acidification. Therefore, nanoparticles remained outside the cells could be clearly distinguished from nanoparticles uptaken by cells as these latter could be spotted inside cellular acidic compartments (such as phagolysosomes, micropinosomes…). Using this model, the endocytosis of 60 nm nanoparticles incubated with the RAW 264.7 macrophages was quantified using time-lapse microscopy and compared to that of 130 nm submicronic particles. The amount of internalized particles was also evaluated by fluorimetry. The biological impact of the particles was also investigated in terms of cytotoxicity, pro-inflammatory response and oxidative stress. Results clearly demonstrated that nanoparticles were more uptaken and more reactive than submicronic particles. Moreover, we validated a method of endocytosis quantification.

  15. Soy flour-derived carbon dots: facile preparation, fluorescence enhancement, and sensitive Fe3+ detection

    NASA Astrophysics Data System (ADS)

    Fang, Liyang; Xu, Qian; Zheng, Xing; Zhang, Weina; Zheng, Jingtang; Wu, Mingbo; Wu, Wenting

    2016-08-01

    Soy flour-derived carbon quantum dots (C-dots) were successfully synthesized via a facile one-step hydrothermal approach. The as-prepared C-dots exhibit an average diameter of 2.5 nm and the crystalline lattices are consistent with graphitic carbons. Meanwhile, they show strong photoluminescence (quantum yield is 7.85 %), good water solubility, and high photostability. Importantly, structural defects of the C-dots were designed to obtain controllable fluorescence, which was achieved by changing the contents of N defects and O defects of C-dots. Our results indicate that N defects can more effectively enhance the fluorescence emission than O defects. As the preparation temperature increases, the N defects are fine-tuned by substituting for partial O defects, reducing nonradiative recombination and enhancing fluorescence intensity, which is further confirmed by surface passivation. Due to its fine photostability, high sensitivity, and good selectivity for Fe3+, the as-prepared C-dots were used as fluorescence probes for detection of ferric ion. The detection limitation comes to 0.021 µM.

  16. TiO2-nanotube-based dye-sensitized solar cells containing fluorescent material.

    PubMed

    Kim, Woong-Rae; Lee, Young-Joon; Park, Hun; Lee, Jae-Joon; Choi, Won-Youl

    2013-05-01

    We fabricated a dye-sensitized solar cells (DSCs) with TiO2 nanotube arrays obtained by anodization of Ti foil. Vertical structure of TiO2 nanotube arrays is very attractive due to a high electron transfer from dye to electrode. To improve the power conversion efficiency, fluorescent material, F-6377, was applied in TiO2-nanotube-based DSCs to use a light spectrum efficiently. Fluorescent material was absorbed the different wavelength of 460 nm from the light absorbed by N719 dye. Fluorescent material to emit the absorbed light energy provided an additional light for dye in DSCs and additional electrons was generated. Thickness of TiO2 nanotube arrays grown by anodic oxidation was 15 microm. N719 dye and 13(-)/l(-) electrolyte were used to fabricate the DSCs. The short circuit current densities (J(sc)) and the power conversion efficiency in DSCs with fluorescent were 10.8 mA/cm2 and 2.48%, respectively. Electrochemical impedance spectroscopy (EIS) was observed to understand an electron transfer and life time.

  17. Fluorescence Based Characterization of Calcium Sensitizer Action on the Troponin Complex

    SciTech Connect

    Schlecht, William; Li, King-Lun; Hu, Dehong; Dong, Wen-Ji

    2016-02-01

    By examining the behavior of each Ca2+ -sensitizer on cTnC at different levels of reconstitution (cTnI-cTnC, full troponin, or full troponin in thin filament) the importance of these proteins on sensitizer efficacy was evaluated, lending insight into the mechanism of action behind each drug. A fluorescence based approach was used to monitor the opening and closing of cardiac troponin C's hydrophobic pocket in the presence and absence of four common Ca2+ -sensitizers: EMD 57033, levosimendan, bepridil and pimobendan. Ca2+ -titration experiments were employed to determine the effect on Ca2+- sensitivity and cooperativity of cTnC opening, while stopped flow experiments were used to investigate the impact on cTnC relaxation kinetics. This study shows EMD 57033 is unable to sensitize cTnC to Ca2+, and likely requires the presence of myosin to illicit a response. Levosimendan, bepridil, and pimobendan were all able to increase the sensitivity of cTnC for Ca2+ to varying degrees; levosimendan and pimobendan reduced the rate of cTnC closing, while bepridil increased this rate. Additionally the same experiments were run on thin filament samples containing cTnT (T204E), a known Ca2+- blunting phosphorylation mimic. Levosimendan, bepridil, and pimobendan were found to elevate the Ca2+-sensitivity of cTnT(T204E) containing thin filaments to within range of the wild type thin filaments.

  18. Ratio-metric sensor to detect riboflavin via fluorescence resonance energy transfer with ultrahigh sensitivity

    NASA Astrophysics Data System (ADS)

    Wang, Jilong; Su, Siheng; Wei, Junhua; Bahgi, Roya; Hope-Weeks, Louisa; Qiu, Jingjing; Wang, Shiren

    2015-08-01

    In this paper, a novel fluorescence resonance energy transfer (FRET) ration-metric fluorescent probe based on heteroatom N, S doped carbon dots (N, S-CDs) was developed to determine riboflavin in aqueous solutions. The ratio of two emission intensities at different wavelengths is applied to determine the concentration of riboflavin (RF). This method is more effective in reducing the background interference and fluctuation of diverse conditions. Therefore, this probe obtains high sensitivity with a low limit of detection (LOD) of 1.9 nM (0.7 ng/ml) which is in the highest level of all riboflavin detection approaches and higher than single wavelength intensity detection (1.9 μM). In addition, this sensor has a high selectivity of detecting riboflavin in deionized water (pH=7) with other biochemical like amino acids. Moreover, riboflavin in aqueous solution is very sensitive to sunlight and can be degraded to lumiflavin, which is toxic. Because the N, S doped carbon dots cannot serve as an energy donor for N, S doped carbon dots and lumiflavin system, this system makes it easy to determine whether the riboflavin is degraded or not, which is first to be reported. This platform may provide possibilities to build a new and facile fluorescence resonance energy transfer based sensor to detect analytes and metamorphous analytes in aqueous solution.

  19. [Fluorescence used to investigate the sensitivity of spinach chloroplast membrane to low intensity electromagnetic radiation].

    PubMed

    Xi, Gang; Yang, Yun-Jing; Lu, Hong

    2009-07-01

    A system for studying biological effect of radio frequency electromagnetic field was developed. The system can form an area where electromagnetic wave with large frequency range is well distributed. The strength of electromagnetic wave was measured easily. Electromagnetic wave in the system did not have effect on environment. The sensitivity of spinach chloroplast membrane to low intensity electromagnetic radiation of 300 MHz under power density of 5 mW x cm(-2) was studied by the spectral analysis method of fluorescence of 8-anilino-1-naphthalene-sulfonic acid (ANS) and the changes in chlorophyll a (Chla) fluorescence parameters of spinach chloroplast membrane. The result showed that the position of spectrum of ANS fluorescence of spinach chloroplast membrane did not change, but the intensity of ANS fluorescence was obviously increased under the action of electromagnetic radiation with power density of 1-5 mW x cm(-2). There was an increase in the intensity of ANS fluorescence with the increase in electromagnetic radiation. The increase of ANS fluorescence of spinach chloroplast membrane showed that low level electromagnetic field induced the decrease in fluidity of chloroplast membrane compared with control experiment. The cause of the change in the fluidity could be related to the polarization of chloroplast membrane under the electromagnetic field. The analysis of Chla fluorescence parameters of spinach chloroplast membrane indicated that low level electromagnetic field of 300 MHz made the fluorescence parameters F0 and F(VI/)F(V) decrease, and F(V)/Fo, Fv/F(m) and deltaF(V)/T increase. It was showed that low level electromagnetic field caused the change of non-active center of photosystem II of spinach chloroplast membrane to active center and the increase in potential active and photochemical efficiency of PSII, and promoted the transmit process of electron in photosynthesis of chloroplast membrane of photosynthesis cell in spinach leaf. The study confirmed

  20. A ratiometric fluorescent nanoprobe based on terbium functionalized carbon dots for highly sensitive detection of an anthrax biomarker.

    PubMed

    Chen, Hao; Xie, Yujie; Kirillov, Alexander M; Liu, Liangliang; Yu, Minghui; Liu, Weisheng; Tang, Yu

    2015-03-25

    A ratiometric fluorescent nanoprobe based on terbium functionalized carbon dots (CDs) was designed to detect dipicolinic acid (DPA) as an anthrax biomarker with high selectivity and sensitivity. CDs were generated by one-step synthesis using an ethylenediaminetetraacetic acid precursor, and served as a scaffold for coordination with Tb(3+) and a fluorescence reference.

  1. High-efficiency fluorescent organic light-emitting devices using sensitizing hosts with a small singlet-triplet exchange energy.

    PubMed

    Zhang, Dongdong; Duan, Lian; Li, Chen; Li, Yilang; Li, Haoyuan; Zhang, Deqiang; Qiu, Yong

    2014-08-06

    Materials with small singlet-triplet splits (ΔEST s) are introduced as sensitizing hosts to excite fluorescent dopants, breaking the trade-off between small ΔEST and high radiative decay rates. A highly efficient orange-fluorescent organic light-emitting diode (OLED) is prepared, showing a maximum external quantum efficiency of 12.2%.

  2. Dual fluorescence sensor for trace oxygen and temperature with unmatched range and sensitivity.

    PubMed

    Baleizão, Carlos; Nagl, Stefan; Schäferling, Michael; Berberan-Santos, Mário N; Wolfbeis, Otto S

    2008-08-15

    An optical dual sensor for oxygen and temperature is presented that is highly oxygen sensitive and covers a broad temperature range. Dual sensing is based on luminescence lifetime measurements. The novel sensor contains two luminescent compounds incorporated into polymer films. The temperature-sensitive dye (ruthenium tris-1,10-phenanthroline) has a highly temperature-dependent luminescence and is incorporated in poly(acrylonitrile) to avoid cross-sensitivity to oxygen. Fullerene C70 was used as the oxygen-sensitive probe owing to its strong thermally activated delayed fluorescence at elevated temperatures that is extremely oxygen sensitive. The cross-sensitivity of C70 to temperature is accounted for by means of the temperature sensor. C70 is incorporated into a highly oxygen-permeable polymer, either ethyl cellulose or organosilica. The two luminescent probes have different emission spectra and decay times, and their emissions can be discriminated using both parameters. Spatially resolved sensing is achieved by means of fluorescence lifetime imaging. The response times of the sensor to oxygen are short. The dual sensor exhibits a temperature operation range between at least 0 and 120 degrees C, and detection limits for oxygen in the ppbv range, operating for oxygen concentrations up to at least 50 ppmv. These ranges outperform all dual oxygen and temperature sensors reported so far. The dual sensor presented in this study is especially appropriate for measurements under extreme conditions such as high temperatures and ultralow oxygen levels. This dual sensor is a key step forward in a number of scientifically or commercially important applications including food packaging, for monitoring of hyperthermophilic microorganisms, in space technology, and safety and security applications in terms of detection of oxygen leaks.

  3. Immobilized fluorescent dyes for sensitive pH measurements on enamel surfaces with fiber optics

    NASA Astrophysics Data System (ADS)

    Rumphorst, A.; Seeger, Stefan; Duschner, H.

    1996-01-01

    Information on the pH directly on surfaces of dental enamel is an important aspect in research on tooth decay. As an alternative to pH-electrodes our approach to the problem is the optical determination of pH by pH sensitive fluorescent dyes immobilized to tooth surfaces. In this study a model for measuring pH either on aminated cellulose substrates or on enamel (in vitro) with a fluorescein type dye is presented. The experimental realization is a fiber optic sensor with a nitrogen-pumped dye laser system and photodiode for the detection of the emitted fluorescence light. The surface pH values in the range between 4 and 7 were derived from the ratios of the excitation bands at 490 nm and 460 nm.

  4. Highly selective, sensitive and fast-responsive fluorescent sensor for Hg2 +

    NASA Astrophysics Data System (ADS)

    Niu, Qingfen; Wu, Xingxing; Li, Tianduo; Cui, Yuezhi; Zhang, Shanshan; Li, Xiaoyan

    2016-06-01

    A phenylamine-oligothiophene-based fluorescent sensor 2TBEA was reported. This sensor exhibited highly selective, sensitive and rapid detection of Hg2 + ion in THF/H2O (7/3, v/v) solution through fluorescence quenching. The detection was unaffected by the coexistence of other competitive metal cations including Na+, K+, Ag+, Ca2 +, Fe3 +, Al3 +, Co2 +, Cu2 +, Ni2 +, Zn2 +, Pb2 +, Cd2 +, Fe2 + and Cr3 +. A1:1 binding ratio for 2TBEA - Hg2 + was demonstrated by Job's plot and mole-ratio curves. The coordination process was chemically reversible with EDTA. The detection limit was evaluated to be as low as 6.164 × 10- 8 M.

  5. Paper-based upconversion fluorescence resonance energy transfer biosensor for sensitive detection of multiple cancer biomarkers

    PubMed Central

    Xu, Sai; Dong, Biao; Zhou, Donglei; Yin, Ze; Cui, Shaobo; Xu, Wen; Chen, Baojiu; Song, Hongwei

    2016-01-01

    A paper-based upconversion fluorescence resonance energy transfer assay device is proposed for sensitive detection of CEA. The device is fabricated on a normal filter paper with simple nano-printing method. Upconversion nanoparticles tagged with specific antibodies are printed to the test zones on the test paper, followed by the introduction of assay antigen. Upconversion fluorescence measurements are directly conducted on the test zones after the antigen-to-antibody reactions. Furthermore, a multi-channel test paper for simultaneous detection of multiple cancer biomarkers was established by the same method and obtained positive results. The device showed high anti-interfere, stability, reproducible and low detection limit (0.89 ng/mL), moreover it is very easy to fabricate and operate, which is a promising prospect for a clinical point-of-care test. PMID:27001460

  6. A novel fluorescent probe for rapid and sensitive detection of hydrogen sulfide in living cells

    NASA Astrophysics Data System (ADS)

    Pan, Jian; Xu, Junchao; Zhang, Youlai; Wang, Liang; Qin, Caiqin; Zeng, Lintao; Zhang, Yue

    2016-11-01

    A novel fluorescent probe for H2S was developed based on a far-red emitting indole-BODIPY, which was decorated with morpholine and 2,4-dinitrobenzenesulfonyl (DNBS) group. This probe showed rapid response (t1/2 = 3 min), high selectivity and sensitivity for H2S with significant colorimetric and fluorescence OFF-ON signals, which was triggered by cleavage of 2,4-dinitrobenzenesulfonyl group. This probe could quantitatively detect the concentrations of H2S ranging from 0 to 60 μM, and the detection of limit was found to be as low as 26 nM. Cell imaging results indicated that the probe could detect and visualize H2S in the living cells.

  7. Shape-parameterized diffuse optical tomography holds promise for sensitivity enhancement of fluorescence molecular tomography

    PubMed Central

    Wu, Linhui; Wan, Wenbo; Wang, Xin; Zhou, Zhongxing; Li, Jiao; Zhang, Limin; Zhao, Huijuan; Gao, Feng

    2014-01-01

    A fundamental approach to enhancing the sensitivity of the fluorescence molecular tomography (FMT) is to incorporate diffuse optical tomography (DOT) to modify the light propagation modeling. However, the traditional voxel-based DOT has been involving a severely ill-posed inverse problem and cannot retrieve the optical property distributions with the acceptable quantitative accuracy and spatial resolution. Although, with the aid of an anatomical imaging modality, the structural-prior-based DOT method with either the hard- or soft-prior scheme holds promise for in vivo acquiring the optical background of tissues, the low robustness of the hard-prior scheme to the segmentation error and inferior performance of the soft-prior one in the quantitative accuracy limit its further application. We propose in this paper a shape-parameterized DOT method for not only effectively determining the regional optical properties but potentially achieving reasonable structural amelioration, lending itself to FMT for comparably improved recovery of fluorescence distribution. PMID:25360379

  8. Fluorescence-based sensing of glucose using engineered glucose/galactose-binding protein: A comparison of fluorescence resonance energy transfer and environmentally sensitive dye labelling strategies

    SciTech Connect

    Khan, Faaizah; Gnudi, Luigi; Pickup, John C.

    2008-01-04

    Fluorescence-based glucose sensors using glucose-binding protein (GBP) as the receptor have employed fluorescence resonance energy transfer (FRET) and environmentally sensitive dyes, but with widely varying sensitivity. We therefore compared signal changes in (a) a FRET system constructed by transglutaminase-mediated N-terminal attachment of Alexa Fluor 488/555 as donor and QSY 7 as acceptor at Cys 152 or 182 mutations with (b) GBP labelled with the environmentally sensitive dye badan at C152 or 182. Both FRET systems had a small maximal fluorescence change at saturating glucose (7% and 16%), badan attached at C152 was associated with a 300% maximal fluorescence increase with glucose, though with badan at C182 there was no change. We conclude that glucose sensing based on GBP and FRET does not produce a larger enough signal change for clinical use; both the nature of the environmentally sensitive dye and its site of conjugation seem important for maximum signal change; badan-GBP152C has a large glucose-induced fluorescence change, suitable for development as a glucose sensor.

  9. Expression of pH-sensitive green fluorescent protein in Arabidopsis thaliana

    NASA Technical Reports Server (NTRS)

    Moseyko, N.; Feldman, L. J.

    2001-01-01

    This is the first report on using green fluorescent protein (GFP) as a pH reporter in plants. Proton fluxes and pH regulation play important roles in plant cellular activity and therefore, it would be extremely helpful to have a plant gene reporter system for rapid, non-invasive visualization of intracellular pH changes. In order to develop such a system, we constructed three vectors for transient and stable transformation of plant cells with a pH-sensitive derivative of green fluorescent protein. Using these vectors, transgenic Arabidopsis thaliana and tobacco plants were produced. Here the application of pH-sensitive GFP technology in plants is described and, for the first time, the visualization of pH gradients between different developmental compartments in intact whole-root tissues of A. thaliana is reported. The utility of pH-sensitive GFP in revealing rapid, environmentally induced changes in cytoplasmic pH in roots is also demonstrated.

  10. Bimodal distribution and fluorescence response of environment-sensitive probes in lipid bilayers.

    PubMed

    Klymchenko, Andrey S; Duportail, Guy; Demchenko, Alexander P; Mély, Yves

    2004-05-01

    A remarkable heterogeneity is often observed in the spectroscopic properties of environment-sensitive fluorescence probes in phospholipid bilayers. To explain its origin, we provided a detailed investigation of the fluorescence excitation and emission spectra of 4'-dimethylamino-3-hydroxyflavone (probe F) in bilayer vesicles with the variations of fatty acid composition, polar heads, temperature, and cholesterol content. Probe F, due to excited-state intramolecular proton transfer, exhibits two bands in emission that are differently sensitive to intermolecular interactions-thereby allowing us to distinguish universal (dipole-dipole) and specific (H-bonding) interactions within the bilayer. Spectroscopic, quenching, and anisotropy data suggest the presence of two forms of probe F at different locations in the bilayer: an H-bond free form located below sn(1)-carbonyls and an H-bonded form located at the polar membrane interface. We provide a quantitative analysis of the distribution of the probe between these two locations as well as the polarity of these locations, and show that both the distribution and the polarity contribute to the probe response. Moreover, analysis of literature data on other environment-sensitive probes (Prodan, Laurdan, Nile Red, NBD lipids, etc.) in lipid bilayers allows us to suggest that the bimodal distribution in the lipid bilayer is probably a general feature of low-polar molecules with polar groups capable of H-bonding interactions.

  11. Monitoring Biophysical Properties of Lipid Membranes by Environment-Sensitive Fluorescent Probes

    PubMed Central

    Demchenko, Alexander P.; Mély, Yves; Duportail, Guy; Klymchenko, Andrey S.

    2009-01-01

    We review the main trends in the development of fluorescence probes to obtain information about the structure, dynamics, and interactions in biomembranes. These probes are efficient for studying the microscopic analogs of viscosity, polarity, and hydration, as well as the molecular order, environment relaxation, and electrostatic potentials at the sites of their location. Progress is being made in increasing the information content and spatial resolution of the probe responses. Multichannel environment-sensitive probes that can distinguish between different membrane physicochemical properties through multiple spectroscopic parameters show considerable promise. PMID:19413953

  12. Monitoring biophysical properties of lipid membranes by environment-sensitive fluorescent probes.

    PubMed

    Demchenko, Alexander P; Mély, Yves; Duportail, Guy; Klymchenko, Andrey S

    2009-05-06

    We review the main trends in the development of fluorescence probes to obtain information about the structure, dynamics, and interactions in biomembranes. These probes are efficient for studying the microscopic analogs of viscosity, polarity, and hydration, as well as the molecular order, environment relaxation, and electrostatic potentials at the sites of their location. Progress is being made in increasing the information content and spatial resolution of the probe responses. Multichannel environment-sensitive probes that can distinguish between different membrane physicochemical properties through multiple spectroscopic parameters show considerable promise.

  13. Thin-film-based sensitivity enhancement for total internal reflection fluorescence live-cell imaging.

    PubMed

    Kim, Kyujung; Cho, Eun-Jin; Huh, Yong-Min; Kim, Donghyun

    2007-11-01

    We investigated experimentally the evanescent field enhancement based on dielectric thin films in total internal reflection microscopy. The sample employed two layers of Al2O3 and SiO2 deposited on an SF10 glass substrate. Field intensity enhancement measured by fluorescent excitation of microbeads relative to that of a control sample without dielectric films was polarization dependent, determined as 4.2 and 2.4 for TE and TM polarizations, respectively, and was in good agreement with numerical results. The thin-film-based field enhancement was also applied to live-cell imaging of quantum dots, which confirmed the sensitivity enhancement qualitatively.

  14. (DNS)C: a fluorescent, environmentally sensitive cytidine derivative for the direct detection of GGG triad sequences.

    PubMed

    Kim, Ki Tae; Kim, Hyun Woo; Moon, Dohyun; Rhee, Young Min; Kim, Byeang Hyean

    2013-09-14

    With the goal of developing a fluorescent nucleoside sensitive to its environment, in this study we synthesized (DNS)C, a novel modified 2'-deoxycytidine bearing a 5-(dimethylamino)naphthalene-1-sulfonyl (dansyl) moiety at the N4 position, and tested its properties in monomeric and oligomeric states. (DNS)C undergoes intramolecular photoinduced electron transfer between its dansyl and cytosine units, resulting in remarkable changes in fluorescence that depend on the choice of solvent. In addition, the fluorescence behavior and thermal stability of oligonucleotides containing (DNS)C are dependent on the nature of the flanking and neighboring bases. Notably, (DNS)C exhibits fluorescence enhancement only in fully matched duplex DNA containing a GGG triad sequence. The environmental sensitivity of (DNS)C can be exploited as a fluorescence tool for monitoring the interactions of DNA with other biomolecules, including DNA, RNA, and proteins.

  15. Fluorescence derivatization method for sensitive chromatographic determination of zidovudine based on the Huisgen reaction.

    PubMed

    Maeda, Yuki; Kishikawa, Naoya; Ohyama, Kaname; Wada, Mitsuhiro; Ikeda, Rie; Kuroda, Naotaka

    2014-08-15

    A novel pre-column fluorescence derivatization method for chromatographic analysis of azide compounds was developed based on the Huisgen reaction, which is a specific cycloaddition reaction between an alkyne and an azide. We designed and synthesized a fluorescent alkyne, 2-(4-ethynylphenyl)-4,5-diphenyl-1H-imidazole (DIB-ET) as a reagent. DIB-ET has a lophine skeleton carrying an alkyne acting as fluorophore and reactive center, respectively. In order to evaluate the practicality of DIB-ET, a high-performance liquid chromatography with fluorescence detection method was developed for the determination of zidovudine as a model of azide compound. Zidovudine could be reacted with DIB-ET in the presence of copper(II) sulfate and L-ascorbic acid as catalysts, and the formed derivative was detected fluorometrically. The proposed method allows sensitive and selective determination of zidovudine in plasma samples with the detection limit of 0.28ngmL(-1) at a S/N=3. Finally, the proposed method could be applied to determine the zidovudine concentration in rat plasma after administration of zidovudine without interference from biological components.

  16. Differential heat sensitivity index in barley cultivars (Hordeum vulgare L.) monitored by chlorophyll a fluorescence OKJIP.

    PubMed

    Oukarroum, Abdallah; El Madidi, Saïd; Strasser, Reto J

    2016-08-01

    The objective of this study was to differentiate the heat tolerance in ten varieties of barley (Hordeum vulgare L.) originating from Morocco. Five modern varieties and five landraces (local varieties) collected at five different geographical localities in the south of Morocco were investigated in the present study. After two weeks of growth, detached leaves were short term exposure to various temperatures (25, 30, 35, 40, and 45 °C) for 10 min in the dark. Two chlorophyll a fluorescence parameters derived from chlorophyll a fluorescence transient (OKJIP) (performance index (PIABS) and relative variable fluorescence at the K-step (VK)) were analysed. Heat treatment had a significant effect on the PIABS and VK at 45 °C treatment and the analysis of variance for PIABS and VK is highly significant between all varieties. The slope of the relationship between logPIABS and VK named heat sensitivity index (HSI) was used to evaluate the thermotolerance of photosystem II (PSII) between the studied barley varieties. According to this approach, barley varieties were screened and ranked for improving heat tolerance. HSI was found to be a new indicator with regard to distinguishing heat tolerance of different barley cultivars.

  17. Sensitive Detection of Phosphorus Deficiency in Plants Using Chlorophyll a Fluorescence.

    PubMed

    Frydenvang, Jens; van Maarschalkerweerd, Marie; Carstensen, Andreas; Mundus, Simon; Schmidt, Sidsel Birkelund; Pedas, Pai Rosager; Laursen, Kristian Holst; Schjoerring, Jan K; Husted, Søren

    2015-09-01

    Phosphorus (P) is a finite natural resource and an essential plant macronutrient with major impact on crop productivity and global food security. Here, we demonstrate that time-resolved chlorophyll a fluorescence is a unique tool to monitor bioactive P in plants and can be used to detect latent P deficiency. When plants suffer from P deficiency, the shape of the time-dependent fluorescence transients is altered distinctively, as the so-called I step gradually straightens and eventually disappears. This effect is shown to be fully reversible, as P resupply leads to a rapid restoration of the I step. The fading I step suggests that the electron transport at photosystem I (PSI) is affected in P-deficient plants. This is corroborated by the observation that differences at the I step in chlorophyll a fluorescence transients from healthy and P-deficient plants can be completely eliminated through prior reduction of PSI by far-red illumination. Moreover, it is observed that the barley (Hordeum vulgare) mutant Viridis-zb(63), which is devoid of PSI activity, similarly does not display the I step. Among the essential plant nutrients, the effect of P deficiency is shown to be specific and sufficiently sensitive to enable rapid in situ determination of latent P deficiency across different plant species, thereby providing a unique tool for timely remediation of P deficiency in agriculture.

  18. A quick and simple FISH protocol with hybridization-sensitive fluorescent linear oligodeoxynucleotide probes.

    PubMed

    Wang, Dan Ohtan; Matsuno, Hitomi; Ikeda, Shuji; Nakamura, Akiko; Yanagisawa, Hiroyuki; Hayashi, Yasunori; Okamoto, Akimitsu

    2012-01-01

    Fluorescence in situ hybridization (FISH) is a powerful tool used in karyotyping, cytogenotyping, cancer diagnosis, species specification, and gene-expression analysis. Although widely used, conventional FISH protocols are cumbersome and time consuming. We have now developed a FISH method using exciton-controlled hybridization-sensitive fluorescent oligodeoxynucleotide (ECHO) probes. ECHO-FISH uses a 25-min protocol from fixation to mounting that includes no stringency washing steps. We use ECHO-FISH to detect both specific DNA and RNA sequences with multicolor probes. ECHO-FISH is highly reproducible, stringent, and compatible with other fluorescent cellular labeling techniques. The resolution allows detection of intranuclear speckles of poly(A) RNA in HeLa cells and dissociated hippocampal primary cultures, and mRNAs in the distal dendrites of hippocampal neurons. We also demonstrate detection of telomeric and centromeric DNA on metaphase mouse chromosomes. The simplicity of the ECHO-FISH method will likely accelerate cytogenetic and gene-expression analysis with high resolution.

  19. High sensitivity automated multiplexed immunoassays using photonic crystal enhanced fluorescence microfluidic system.

    PubMed

    Tan, Yafang; Tang, Tiantian; Xu, Haisheng; Zhu, Chenqi; Cunningham, Brian T

    2015-11-15

    We demonstrate a platform that integrates photonic crystal enhanced fluorescence (PCEF) detection of a surface-based microspot fluorescent assay with a microfluidic cartridge to achieve simultaneous goals of high analytic sensitivity (single digit pg/mL), high selectivity, low sample volume, and assay automation. The PC surface, designed to provide optical resonances for the excitation wavelength and emission wavelength of Cyanines 5 (Cy5), was used to amplify the fluorescence signal intensity measured from a multiplexed biomarker microarray. The assay system is comprised of a plastic microfluidic cartridge for holding the PC and an assay automation system that provides a leak-free fluid interface during introduction of a sequence of fluids under computer control. Through the use of the assay automation system and the PC embedded within the microfluidic cartridge, we demonstrate pg/mL-level limits of detection by performing representative biomarker assays for interleukin 3 (IL3) and Tumor Necrosis Factor (TNF-α). The results are consistent with limits of detection achieved without the use of the microfluidic device with the exception that coefficients of variability from spot-to-spot are substantially lower than those obtained by performing assays with manual manipulation of assay liquids. The system's capabilities are compatible with the goal of diagnostic instruments for point-of-care settings.

  20. High Sensitivity Automated Multiplexed Immunoassays Using Photonic Crystal Enhanced Fluorescence Microfluidic System

    PubMed Central

    Tan, Yafang; Tang, Tiantian; Xu, Haisheng; Zhu, Chenqi; Cunningham, Brian T.

    2015-01-01

    We demonstrate a platform that integrates photonic crystal enhanced fluorescence (PCEF) detection of a surface-based microspot fluorescent assay with a microfluidic cartridge to achieve simultaneous goals of high analytic sensitivity (single digit pg/mL), high selectivity, low sample volume, and assay automation. The PC surface, designed to provide optical resonances for the excitation wavelength and emission wavelength of Cyanines 5 (Cy5), was used to amplify the fluorescence signal intensity measured from a multiplexed biomarker microarray. The assay system is comprised of a plastic microfluidic cartridge for holding the PC and an assay automation system that provides a leak-free fluid interface during introduction of a sequence of fluids under computer control. Through the use of the assay automation system and the PC embedded within the microfluidic cartridge, we demonstrate pg/mL-level limits of detection by performing representative biomarker assays for interleukin 3 (IL3) and Tumor Necrosis Factor (TNF-α). The results are consistent with limits of detection achieved without the use of the microfluidic device with the exception that coefficients of variability from spot-to-spot are substantially lower than those obtained by performing assays with manual manipulation of assay liquids. The system’s capabilities are compatible with the goal of diagnostic instruments for point-of-care settings. PMID:26043313

  1. Microanalysis of metals in coal and coal ash using the Stanford/USGS SHRIMP-RG ion microprobe[Sensitive High-Resolution Ion MicroProbe--Reversed Geometry

    SciTech Connect

    Kolker, A.; Zielinski, R.A.; Wooden, J.L.; Persing, H.M.

    2000-07-01

    The capability of the SHRIMP-RG ion microprobe to determine the micro-distribution of selected trace metals in coal and coal ash was investigated as part of a larger study of the behavior of air toxic metals during coal combustion. Initial work, reported here, used the oxygen (O) ion source for in-situ determination of Cr and other elements in illite/smectite, a major inorganic constituent of the coals analyzed. This was followed by tests of the applicability of the SHRIMP-RG for trace-metal analysis of fly ash from a Kentucky power plant, in which U and Pb concentrations were determined in the coarse (63--150 micrometer) fraction of the fly ash. The results for illite/smectite confirm that it is an important source of chromium that may be emitted during coal burning. Results for fly-ash show that the {sup 75}As peak is resolvable from potential interferences in glass standards and partially resolvable in the fly ash, indicating that the SHRIMP-RG may be useful in characterizing the distribution of leachable metals condensed on fly ash surfaces.

  2. Fluorescent Probe Encapsulated in Avidin Protein to Eliminate Nonspecific Fluorescence and Increase Detection Sensitivity in Blood Serum.

    PubMed

    Wu, Ting-Wei; Lee, Fang-Hong; Gao, Ruo-Cing; Chew, Chee Ying; Tan, Kui-Thong

    2016-08-16

    Quantitative detection of trace amounts of a biomarker in protein rich human blood plasma using fluorescent probes is a great challenge as the real signal is usually obscured by nonspecific fluorescence. This problem occurs because most of the fluorescent dyes bind very tightly with blood proteins to produce a large fluorescence increase, resulting in overestimation of the biomarker concentrations and false positive diagnosis. In this paper, we report that biotinylated fluorescent probes encapsulated in avidin protein can generate very specific fluorescence in blood serum by blocking out nonspecific dye-protein interactions. We applied our novel probe design to detect two different types of biomolecules, hydrogen sulfide and nitroreductase. Our Avidin conjugated probes achieved quantitative analyte detection in blood serum; whereas concentrations were overestimated up to 320-fold when bare fluorescent probes were employed. As compared to conventional approaches where fluorescent probes are encapsulated into polymers and nanoparticles, our simple approach successfully overcomes many key issues such as dye leakage, long preparation steps, inconsistent dye-host ratios, difficulty in constructing in situ in a complex medium, and limited application to detect only small metabolites.

  3. Design and performance of a microchip electrophoresis instrument with sensitive variable-wavelength fluorescence detection.

    PubMed

    Belder, Detlev; Deege, Alfred; Maass, Martin; Ludwig, Martin

    2002-07-01

    A modular instrument for high-speed microchip electrophoresis (MCE) equipped with a sensitive variable-wavelength fluorescence detection system was developed and evaluated. The experimental setup consists mainly of a lamp-based epifluorescence microscope for variable-wavelength fluorescence detection and imaging and a programmable four-channel bipolar high-voltage source capable of delivering up to +/- 10 kV per channel. The optical unit was equipped with a high-sensitivity photomultiplier tube and an adjustable aperture. The system was applied to MCE separations of flurescein isothiocyanate (FITC)-labelled amines utilizing blue light (450-480 nm) for excitation as well as for the separation of rhodamines utilizing excitation light in the green spectral region (531-560 nm). At optimized conditions baseline separation of four FITC-labelled amines could be obtained in less than 50 s at a detection limit of 460 ppt (1 nM) with a signal-to-noise ratio of 3:1. Three rhodamines could be baseline-separated in less than 6 s at a detection limit of 240 ppt (500 pM). The relative standard deviations of absolute migration times determined in repetitive MCE separations of FITC-labelled amines were below 2.5% (n= 25). By the application of cyclodextrin-modified electrolytes, chiral separation of FITC-labelled amines could be performed in seconds demonstrating the potential of microchip electrophoresis for chiral high-throughput screening.

  4. Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging.

    PubMed

    Afsari, Hamid Samareh; Cardoso Dos Santos, Marcelina; Lindén, Stina; Chen, Ting; Qiu, Xue; van Bergen En Henegouwen, Paul M P; Jennings, Travis L; Susumu, Kimihiro; Medintz, Igor L; Hildebrandt, Niko; Miller, Lawrence W

    2016-06-01

    Time-gated Förster resonance energy transfer (FRET) using the unique material combination of long-lifetime terbium complexes (Tb) and semiconductor quantum dots (QDs) provides many advantages for highly sensitive and multiplexed biosensing. Although time-gated detection can efficiently suppress sample autofluorescence and background fluorescence from directly excited FRET acceptors, Tb-to-QD FRET has rarely been exploited for biomolecular imaging. We demonstrate Tb-to-QD time-gated FRET nanoassemblies that can be applied for intra- and extracellular imaging. Immunostaining of different epitopes of the epidermal growth factor receptor (EGFR) with Tb- and QD-conjugated antibodies and nanobodies allowed for efficient Tb-to-QD FRET on A431 cell membranes. The broad usability of Tb-to-QD FRET was further demonstrated by intracellular Tb-to-QD FRET and Tb-to-QD-to-dye FRET using microinjection as well as cell-penetrating peptide-mediated endocytosis with HeLa cells. Effective brightness enhancement by FRET from several Tb to the same QD, the use of low nanomolar concentrations, and the quick and sensitive detection void of FRET acceptor background fluorescence are important advantages for advanced intra- and extracellular imaging of biomolecular interactions.

  5. Time-gated FRET nanoassemblies for rapid and sensitive intra- and extracellular fluorescence imaging

    PubMed Central

    Afsari, Hamid Samareh; Cardoso Dos Santos, Marcelina; Lindén, Stina; Chen, Ting; Qiu, Xue; van Bergen en Henegouwen, Paul M. P.; Jennings, Travis L.; Susumu, Kimihiro; Medintz, Igor L.; Hildebrandt, Niko; Miller, Lawrence W.

    2016-01-01

    Time-gated Förster resonance energy transfer (FRET) using the unique material combination of long-lifetime terbium complexes (Tb) and semiconductor quantum dots (QDs) provides many advantages for highly sensitive and multiplexed biosensing. Although time-gated detection can efficiently suppress sample autofluorescence and background fluorescence from directly excited FRET acceptors, Tb-to-QD FRET has rarely been exploited for biomolecular imaging. We demonstrate Tb-to-QD time-gated FRET nanoassemblies that can be applied for intra- and extracellular imaging. Immunostaining of different epitopes of the epidermal growth factor receptor (EGFR) with Tb- and QD-conjugated antibodies and nanobodies allowed for efficient Tb-to-QD FRET on A431 cell membranes. The broad usability of Tb-to-QD FRET was further demonstrated by intracellular Tb-to-QD FRET and Tb-to-QD-to-dye FRET using microinjection as well as cell-penetrating peptide–mediated endocytosis with HeLa cells. Effective brightness enhancement by FRET from several Tb to the same QD, the use of low nanomolar concentrations, and the quick and sensitive detection void of FRET acceptor background fluorescence are important advantages for advanced intra- and extracellular imaging of biomolecular interactions. PMID:27386579

  6. A highly sensitive ratiometric fluorescent probe for the detection of cytoplasmic and nuclear hydrogen peroxide.

    PubMed

    Wen, Ying; Liu, Keyin; Yang, Huiran; Li, Yi; Lan, Haichuang; Liu, Yi; Zhang, Xinyu; Yi, Tao

    2014-10-07

    As a marker for oxidative stress and a second messenger in signal transduction, hydrogen peroxide (H2O2) plays an important role in living systems. It is thus critical to monitor the changes in H2O2 in cells and tissues. Here, we developed a highly sensitive and versatile ratiometric H2O2 fluorescent probe (NP1) based on 1,8-naphthalimide and boric acid ester. In response to H2O2, the ratio of its fluorescent intensities at 555 and 403 nm changed 1020-fold within 200 min. The detecting limit of NP1 toward H2O2 is estimated as 0.17 μM. It was capable of imaging endogenous H2O2 generated in live RAW 264.7 macrophages as a cellular inflammation response, and especially, it was able to detect H2O2 produced as a signaling molecule in A431 human epidermoid carcinoma cells through stimulation by epidermal growth factor. This probe contains an azide group and thus has the potential to be linked to various molecules via the click reaction. After binding to a Nuclear Localization Signal peptide, the peptide-based combination probe (pep-NP1) was successfully targeted to nuclei and was capable of ratiometrically detecting nuclear H2O2 in living cells. These results indicated that NP1 was a highly sensitive ratiometric H2O2 dye with promising biological applications.

  7. A direct, continuous, sensitive assay for protein disulphide-isomerase based on fluorescence self-quenching.

    PubMed

    Raturi, Arun; Vacratsis, Panayiotis O; Seslija, Dana; Lee, Lana; Mutus, Bulent

    2005-10-15

    PDI (protein disulphide-isomerase) activity is generally monitored by insulin turbidity assay or scrambled RNase assay, both of which are performed by UV-visible spectroscopy. In this paper, we present a sensitive fluorimetric assay for continuous determination of disulphide reduction activity of PDI. This assay utilizes the pseudo-substrate diabz-GSSG [where diabz stands for di-(o-aminobenzoyl)], which is formed by the reaction of isatoic anhydride with the two free N-terminal amino groups of GSSG. The proximity of two benzoyl groups leads to quenching of the diabz-GSSG fluorescence by approx. 50% in comparison with its non-disulphide-linked form, abz-GSH (where abz stands for o-aminobenzoyl). Therefore the PDI-dependent disulphide reduction can be monitored by the increase in fluorescence accompanying the loss of proximity-quenching upon conversion of diabz-GSSG into abz-GSH. The apparent K(m) of PDI for diabz-GSSG was estimated to be approx. 15 muM. Unlike the insulin turbidity assay and scrambled RNase assay, the diabz-GSSG-based assay was shown to be effective in determining a single turnover of enzyme in the absence of reducing agents with no appreciable blank rates. The assay is simple to perform and very sensitive, with an estimated detection limit of approx. 2.5 nM PDI, enabling its use for the determination of platelet surface PDI activity in crude sample preparations.

  8. Design of a sensitive fluorescent polarization immunoassay for rapid screening of milk for cephalexin.

    PubMed

    Beloglazova, Natalia V; Eremin, Sergei A

    2015-11-01

    In this paper we describe the development of a sensitive, fast, and easily performed fluorescence polarization immunoassay for determination of cephalexin in milk. The experimental work was performed to increase sensitivity and specificity. Therefore, the structures of the tracers were varied by synthesis of both cephalexin (CEX) and cephalotin (CET) conjugates with a variety of fluorescent labels. Two rabbit antisera containing antibodies against cephalexin and cephalotin were tested in homologous and heterologous combinations with the tracers. For every working antibody-tracer combination, the analytical conditions and cross-reactivity for structural analogues-cephalosporins and other antibiotics that could also be present in milk-were determined. It was found that the highest sensitivity was achieved by use of the homologous pair CET-EDF-anti-CET antibody (limit of detection (LOD) 0.4 μg kg(-1) for standard solutions prepared in buffer), but this combination was not appropriate because of high cross-reactivity with CET. For subsequent experiments, therefore, CEX- EDF-anti-CEX antibody were chosen (LOD 0.8 μg kg(-1) for standard solutions prepared in buffer). Part of this manuscript is devoted to the variation of precipitation agents for pretreatment of milk before analysis; milk is an extremely complicated matrix. The optimum protein precipitation agent was methanol. This technique for cephalexin determination was characterized by a limit of detection of 1 μg kg(-1). The method was validated by using naturally contaminated and spiked milk samples. The results obtained corresponded very well with those obtained by HPLC, which was used as confirmation method.

  9. Photo- and biophysical studies of lectin-conjugated fluorescent nanoparticles: reduced sensitivity in high density assays.

    PubMed

    Wang, Yaqi; Gildersleeve, Jeffrey C; Basu, Amit; Zimmt, Matthew B

    2010-11-18

    Lectin-conjugated, fluorescent silica nanoparticles (fNP) have been developed for carbohydrate-based histopathology evaluations of epithelial tissue biopsies. The fNP platform was selected for its enhanced emissive brightness compared to direct dye labeling. Carbohydrate microarray studies were performed to compare the carbohydrate selectivity of the mannose-recognizing lectin Concanavalin A (ConA) before and after conjugation to fluorescent silica nanoparticles (ConA-fNP). These studies revealed surprisingly low emission intensities upon staining with ConA-fNP compared to those with biotin-ConA/Cy3-streptavidin staining. A series of photophysical and biophysical characterizations of the fNP and ConA-fNP conjugates were performed to probe the low sensitivity from fNP in the microarray assays. Up to 1200 fluorescein (FL) and 80 tetramethylrhodamine (TR) dye molecules were incorporated into 46 nm diameter fNP, yielding emissive brightness values 400 and 35 times larger than the individual dye molecules, respectively. ConA lectin conjugated to carboxylic acid surface-modified nanoparticles covers 15-30% of the fNP surface. The CD spectra and mannose substrate selectivity of ConA conjugated to the fNP differed slightly compared to that of soluble ConA. Although, the high emissive brightness of fNP enhances detection sensitivity for samples with low analyte densities, large fNP diameters limit fNP recruitment and binding to samples with high analyte densities. The high analyte density and nearly two-dimensional target format of carbohydrate microarrays make probe size a critical parameter. In this application, fNP labels afford minimal sensitivity advantage compared to direct dye labeling.

  10. Optimized Detector Angular Configuration Increases the Sensitivity of X-ray Fluorescence Computed Tomography (XFCT).

    PubMed

    Ahmad, Moiz; Bazalova-Carter, Magdalena; Fahrig, Rebecca; Xing, Lei

    2015-05-01

    In this work, we demonstrated that an optimized detector angular configuration based on the anisotropic energy distribution of background scattered X-rays improves X-ray fluorescence computed tomography (XFCT) detection sensitivity. We built an XFCT imaging system composed of a bench-top fluoroscopy X-ray source, a CdTe X-ray detector, and a phantom motion stage. We imaged a 6.4-cm-diameter phantom containing different concentrations of gold solution and investigated the effect of detector angular configuration on XFCT image quality. Based on our previous theoretical study, three detector angles were considered. The X-ray fluorescence detector was first placed at 145 (°) (approximating back-scatter) to minimize scatter X-rays. XFCT image quality was compared to images acquired with the detector at 60 (°) (forward-scatter) and 90 (°) (side-scatter). The datasets for the three different detector positions were also combined to approximate an isotropically arranged detector. The sensitivity was optimized with detector in the 145 (°) back-scatter configuration counting the 78-keV gold Kβ1 X-rays. The improvement arose from the reduced energy of scattered X-ray at the 145 (°) position and the large energy separation from gold K β1 X-rays. The lowest detected concentration in this configuration was 2.5 mgAu/mL (or 0.25% Au with SNR = 4.3). This concentration could not be detected with the 60 (°) , 90 (°) , or isotropic configurations (SNRs = 1.3, 0, 2.3, respectively). XFCT imaging dose of 14 mGy was in the range of typical clinical X-ray CT imaging doses. To our knowledge, the sensitivity achieved in this experiment is the highest in any XFCT experiment using an ordinary bench-top X-ray source in a phantom larger than a mouse ( > 3 cm).

  11. pHuji, a pH-sensitive red fluorescent protein for imaging of exo- and endocytosis

    PubMed Central

    Shen, Yi; Rosendale, Morgane

    2014-01-01

    Fluorescent proteins with pH-sensitive fluorescence are valuable tools for the imaging of exocytosis and endocytosis. The Aequorea green fluorescent protein mutant superecliptic pHluorin (SEP) is particularly well suited to these applications. Here we describe pHuji, a red fluorescent protein with a pH sensitivity that approaches that of SEP, making it amenable for detection of single exocytosis and endocytosis events. To demonstrate the utility of the pHuji plus SEP pair, we perform simultaneous two-color imaging of clathrin-mediated internalization of both the transferrin receptor and the β2 adrenergic receptor. These experiments reveal that the two receptors are differentially sorted at the time of endocytic vesicle formation. PMID:25385186

  12. Enhancement of fluorescence development of end products by use of a fluorescence developer solution in a rapid and sensitive fluorescent spot test for specific detection of microbial beta-lactamases.

    PubMed

    Chen, K C; Holmes, K K

    1986-03-01

    A fluorescent spot test method for specific detection of microbial beta-lactamases as previously published (K. C. S. Chen, J. S. Knapp, and K. K. Holmes, J. Clin. Microbiol. 19:818-825, 1984) was improved by the use of a fluorescence developer solution. The fluorescence developer solution used in this study consisted of 0.78 M sodium tartrate buffer containing 12% formaldehyde at a final pH of 4.5. An addition of 1 volume of fluorescence developer solution to 5 volumes of ampicillin or cephalex substrate solution incubated with beta-lactamase-producing organisms, followed by heating the mixture at 45 degrees C for 10 min resulted in enhancement of fluorescence of the end products of beta-lactamase activity. This provides a more sensitive assay for microbial beta-lactamases and offers the potential for direct detection of beta-lactamases in clinical specimens.

  13. [Development of novel fluorescence-derivatization-HPLC methods enabling highly sensitive and selective analysis of biological compounds].

    PubMed

    Todoroki, Kenichiro

    2011-01-01

    Fluorescence-derivatization-HPLC methods are powerful tools for performing the analysis of bioactive compounds with high sensitivity and selectivity. In this paper, the author reviews the development of the following four types of novel fluorescence-derivatization-HPLC analytical systems: (1) simultaneous HPLC analysis of melatonin and its related compounds through post-column electrochemical demethylation and fluorescence derivatization, (2) HPLC analysis of 5-hydroxyindoles based on fluorescence derivatization by online pre-column photocatalytic oxidation with benzylamine, (3) reagent peak-free HPLC analysis for aliphatic amines and amino acids using F-trap pyrene as a fluorous tag-bound fluorescence derivatization reagent, and (4) reagent peak-free HPLC analysis for carboxylic acids using a fluorous scavenging-derivatization method. The authors have also successfully applied these systems to biological and pharmaceutical analyses.

  14. Highly sensitive fluorescence quantitative detection of specific DNA sequences with molecular beacons and nucleic acid dye SYBR Green I.

    PubMed

    Xiang, Dongshan; Zhai, Kun; Xiang, Wenjun; Wang, Lianzhi

    2014-11-01

    A highly sensitive fluorescence method of quantitative detection for specific DNA sequence is developed based on molecular beacon (MB) and nucleic acid dye SYBR Green I by synchronous fluorescence analysis. It is demonstrated by an oligonucleotide sequence of wild-type HBV (target DNA) as a model system. In this strategy, the fluorophore of MB is designed to be 6-carboxyfluorescein group (FAM), and the maximum excitation wavelength and maximum emission wavelength are both very close to that of SYBR Green I. In the presence of targets DNA, the MBs hybridize with the targets DNA and form double-strand DNA (dsDNA), the fluorophore FAM is separated from the quencher BHQ-1, thus the fluorophore emit fluorescence. At the same time, SYBR Green I binds to dsDNA, the fluorescence intensity of SYBR Green I is significantly enhanced. When targets DNA are detected by synchronous fluorescence analysis, the fluorescence peaks of FAM and SYBR Green I overlap completely, so the fluorescence signal of system will be significantly enhanced. Thus, highly sensitive fluorescence quantitative detection for DNA can be realized. Under the optimum conditions, the total fluorescence intensity of FAM and SYBR Green I exhibits good linear dependence on concentration of targets DNA in the range from 2×10(-11) to 2.5×10(-9)M. The detection limit of target DNA is estimated to be 9×10(-12)M (3σ). Compared with previously reported methods of detection DNA with MB, the proposed method can significantly enhance the detection sensitivity.

  15. Highly sensitive ratiometric detection of heparin and its oversulfated chondroitin sulfate contaminant by fluorescent peptidyl probe.

    PubMed

    Mehta, Pramod Kumar; Lee, Hyeri; Lee, Keun-Hyeung

    2017-05-15

    The selective and sensitive detection of heparin, an anticoagulant in clinics as well as its contaminant oversulfated chondroitin sulfate (OSCS) is of great importance. We first reported a ratiometric sensing method for heparin as well as OSCS contaminants in heparin using a fluorescent peptidyl probe (Pep1, pyrene-GSRKR) and heparin-digestive enzyme. Pep1 exhibited a highly sensitive ratiometric response to nanomolar concentration of heparin in aqueous solution over a wide pH range (2~11) and showed highly selective ratiometric response to heparin among biological competitors such as hyaluronic acid and chondroitin sulfate. Pep1 showed a linear ratiometric response to nanomolar concentrations of heparin in aqueous solutions and in human serum samples. The detection limit for heparin was calculated to be 2.46nM (R(2)=0.99) in aqueous solutions, 2.98nM (R(2)=0.98) in 1% serum samples, and 3.43nM (R(2)=0.99) in 5% serum samples. Pep1 was applied to detect the contaminated OSCS in heparin with heparinase I, II, and III, respectively. The ratiometric sensing method using Pep1 and heparinase II was highly sensitive, fast, and efficient for the detection of OSCS contaminant in heparin. Pep1 with heparinase II could detect as low as 0.0001% (w/w) of OSCS in heparin by a ratiometric response.

  16. Improving sensitivity of gold nanoparticle based fluorescence quenching and colorimetric aptasensor by using water resuspended gold nanoparticle.

    PubMed

    Liu, Jinchuan; Guan, Zheng; Lv, Zhenzhen; Jiang, Xiaoling; Yang, Shuming; Chen, Ailiang

    2014-02-15

    Gold nanoparticles (AuNPs) based fluorescence quenching or colorimetric aptasensor have been developed for many analytes recently largely because of the ease of detection, high sensitivity, and potential for high-throughput analysis. However, the effects of remnant non-AuNPs components in the colloid gold solution on these assays performance remain unclear. For the first time, we demonstrated that the remnant sodium citrate and the reaction products of three acids play counteractive roles in AuNPs based fluorescence quenching and colorimetric aptasensor in three ways in this study. First, the remnant sodium citrate in the colloid gold solution could increase the fluorescence intensity of FAM labeled on the aptamer that reduce the efficiency of AuNPs fluorescent quenching. Second, the reaction products of citric acid, HCl and ketoglutaric acid reduce the fluorescence recovery by quenching the fluorescence of FAM labeled on the aptamer dissociated from the surface of AuNPs upon addition of target. Lastly, the reaction products of three acids reduce the pH value of the colloid gold solution that reduce the sensitivity of AuNPs based colorimetric aptasensor by increasing the adsorption of aptamer to surface of AuNPs. With sulfadimethoxine and thrombin as model analytes, we found that water resuspended AuNPs can significantly increase the sensitivity by more than 10-fold for AuNPs based fluorescence quenching aptasensor. In the AuNPs based colorimetric aptasensor for sulfadimethoxine using the water resuspended AuNPs, the sensitivity also was increased by 10-fold compared with that of original AuNPs. The findings in this study provide theoretical guidance for further improving AuNPs based fluorescent quenching and colorimetric aptasensor by adjusting the composition of AuNPs solution.

  17. Green synthesis of carbon nanodots as an effective fluorescent probe for sensitive and selective detection of mercury(II) ions

    NASA Astrophysics Data System (ADS)

    Lu, Wenbo; Qin, Xiaoyun; Asiri, Abdullah M.; Al-Youbi, Abdulrahman O.; Sun, Xuping

    2013-01-01

    The present communication reports on the use of sweet potatoes as carbon source for green synthesis of fluorescent carbon nanodots (CNDs) ranging from 1 to 3 nm. We further demonstrate the use of such CNDs as a very effective fluorescent probe for label-free, sensitive, and selective detection of Hg2+ with a detection limit as low as 1 nM. The feasibility of the CNDs for analysis of Hg2+ in a real water sample is also demonstrated successfully.

  18. A sensitive fluorescence method for detection of E. Coli using rhodamine 6G dyeing.

    PubMed

    Wang, Yaohui; Jiang, Caina; Wen, Guiqing; Zhang, Xinghui; Luo, Yanghe; Qin, Aimiao; Liang, Aihui; Jiang, Zhiliang

    2016-06-01

    Negatively charged bacteria combined with positively charged alkaline dye rhodamine 6G (Rh6G) in NaH2 PO4 -Na2 HPO4 buffer solution pH 7.4, by electrostatic interaction. The dyed bacteria exhibited a strong fluorescence peak at 552 nm and fluorescence intensity was directly linear to Escherichia coli (E. coli), Bacillus subtilis (B. subtilis) and Staphylococcus aureus (S. aureus) concentrations in the range of 7.06 × 10(4) to 3.53 × 10(7) , 4.95 × 10(5) to 2.475 × 10(8) and 32.5 to 16250 colony forming unit/mL (cfu/mL) respectively, with detection limits of 3.2 × 10(4) cfu/mL E. coli, 2.3 × 10(5) cfu/mL B. subtilis and 16 cfu/mL S. aureus, respectively. Samples were cultured for 12 h, after which the linear detection range for E. coli was 2 to 88 cfu/mL. This simple, rapid and sensitive method was used for the analysis of water and drinking samples. Copyright © 2015 John Wiley & Sons, Ltd.

  19. Highly sensitive immunoassay of protein molecules based on single nanoparticle fluorescence detection in a nanowell

    NASA Astrophysics Data System (ADS)

    Han, Jin-Hee; Kim, Hee-Joo; Lakshmana, Sudheendra; Gee, Shirley J.; Hammock, Bruce D.; Kennedy, Ian M.

    2011-03-01

    A nanoarray based-single molecule detection system was developed for detecting proteins with extremely high sensitivity. The nanoarray was able to effectively trap nanoparticles conjugated with biological sample into nanowells by integrating with an electrophoretic particle entrapment system (EPES). The nanoarray/EPES is superior to other biosensor using immunoassays in terms of saving the amounts of biological solution and enhancing kinetics of antibody binding due to reduced steric hindrance from the neighboring biological molecules. The nanoarray patterned onto a layer of PMMA and LOL on conductive and transparent indium tin oxide (ITO)-glass slide by using e-beam lithography. The suspension of 500 nm-fluorescent (green emission)-carboxylated polystyrene (PS) particles coated with protein-A followed by BDE 47 polyclonal antibody was added to the chip that was connected to the positive voltage. The droplet was covered by another ITO-coated-glass slide and connected to a ground terminal. After trapping the particles into the nanowells, the solution of different concentrations of anti-rabbit- IgG labeled with Alexa 532 was added for an immunoassay. A single molecule detection system could quantify the anti-rabbit IgG down to atto-mole level by counting photons emitted from the fluorescent dye bound to a single nanoparticle in a nanowell.

  20. Highly selective and sensitive fluorescent sensor: Thiacalix[4]arene-1-naphthalene carboxylate for Zn2+ ions

    NASA Astrophysics Data System (ADS)

    Darjee, Savan M.; Modi, Krunal M.; Panchal, Urvi; Patel, Chirag; Jain, Vinod K.

    2017-04-01

    Thiacalix[4]arene based fluorescent sensor bearing two naphthoyl groups, thiacalix-1-naphthalene carboxylate (TCNC) has been synthesized and characterized by 1H NMR, 13C NMR, FTIR, ESI-MS spectroscopic techniques. The interaction behavior of TCNC with various metal ions like Fe3+, Hg2+, Co2+, Ni2+, Cu2+, Cd2+, Pb2+, Mg2+, K+, Na+, and Zn2+ was studied by UV-visible and emission spectrophotometry. It was observed that TCNC recognizes Zn2+ ions with high selectivity and sensitivity. The enhancement of fluorescence intensity due to presence of Zn2+ ions was not perturbed in the presence of high concentration of other associated metal ions. The 1:1 stoichiometry of TCNC:Zn2+ complex was confirmed by job's plot, ESI-MS study and 1H NMR titration. The binding constant and quantum yield were also calculated by using spectrofluorimetric titration data. Linear detection range of zinc ions was found to be 1 nM-740 nM. Furthermore, molecular docking study was performed to evaluate the binding affinity and possible interactions between TCNC and Zn2+ depicting that TCNC interact with Zn2+ via weak intramolecular forces. In addition to that molecular dynamics has also been performed to evaluate the conformational changes and it's structural stability in the particular environment.

  1. High-sensitivity detection of breast tumors in vivo by use of a pH-sensitive near-infrared fluorescence probe

    NASA Astrophysics Data System (ADS)

    Mathejczyk, Julia Eva; Pauli, Jutta; Dullin, Christian; Resch-Genger, Ute; Alves, Frauke; Napp, Joanna

    2012-07-01

    We investigated the potential of the pH-sensitive dye, CypHer5E, conjugated to Herceptin (pH-Her) for the sensitive detection of breast tumors in mice using noninvasive time-domain near-infrared fluorescence imaging and different methods of data analysis. First, the fluorescence properties of pH-Her were analyzed as function of pH and/or dye-to-protein ratio, and binding specificity was confirmed in cell-based assays. Subsequently, the performance of pH-Her in nude mice bearing orthotopic HER2-positive (KPL-4) and HER2-negative (MDA-MB-231) breast carcinoma xenografts was compared to that of an always-on fluorescent conjugate Alexa Fluor 647-Herceptin (Alexa-Her). Subtraction of autofluorescence and lifetime (LT)-gated image analyses were performed for background fluorescence suppression. In mice bearing HER2-positive tumors, autofluorescence subtraction together with the selective fluorescence enhancement of pH-Her solely in the tumor's acidic environment provided high contrast-to-noise ratios (CNRs). This led to an improved sensitivity of tumor detection compared to Alexa-Her. In contrast, LT-gated imaging using LTs determined in model systems did not improve tumor-detection sensitivity in vivo for either probe. In conclusion, pH-Her is suitable for sensitive in vivo monitoring of HER2-expressing breast tumors with imaging in the intensity domain and represents a promising tool for detection of weak fluorescent signals deriving from small tumors or metastases.

  2. Resolution of heterogeneous fluorescence emission signals and decay lifetime measurement on fluorochrome-labeled cells by phase-sensitive FCM

    SciTech Connect

    Steinkamp, J.A.; Crissman, H.A.

    1993-02-01

    A phase-sensitive flow cytometer has been developed to resolve signals from heterogeneous fluorescence emission spectra and quantify fluorescence decay times on cells labeled with fluorescent dyes. This instrument combines flow cytometry (FCM) and fluorescence spectroscopy measurement principles to provide unique capabilities for making phase-resolved measurements on single cells in flow, while preserving conventional FCM measurement capabilities. Stained cells are analyzed as they pass through an intensity-modulated (sinusoid) laser excitation beam. Fluorescence is measured orthogonally using a s barrier filter to block scattered laser excitation light, and a photomultiplier tube detector output signals, which are shifted in phase from a reference signal and amplitude demodulated, are processed by phase-sensitive detection electronics to resolve signals from heterogeneous emissions and quantify decay lifetimes directly. The output signals are displayed as frequency distribution histograms and bivariate diagrams using a computer-based data acquisition system. Results have demonstrated signal phase shift, amplitude demodulation, and average measurement of fluorescence lifetimes on stained cells; a detection limit threshold of 300 to 500 fluorescein isothiocyanate (FITC); fluorescence measurement precision of 1.3% on alignment fluorospheres and 3.4% on propidium iodide (PI)-stained cells; the resolution of PI and FITC signals from cells stainedin combination with PI and FITC, based on differences in their decay lifetimes; and the ability to measure single decay nines by the two-phase, phase comparator, method.

  3. Resolution of heterogeneous fluorescence emission signals and decay lifetime measurement on fluorochrome-labeled cells by phase-sensitive FCM

    SciTech Connect

    Steinkamp, J.A.; Crissman, H.A.

    1993-01-01

    A phase-sensitive flow cytometer has been developed to resolve signals from heterogeneous fluorescence emission spectra and quantify fluorescence decay times on cells labeled with fluorescent dyes. This instrument combines flow cytometry (FCM) and fluorescence spectroscopy measurement principles to provide unique capabilities for making phase-resolved measurements on single cells in flow, while preserving conventional FCM measurement capabilities. Stained cells are analyzed as they pass through an intensity-modulated (sinusoid) laser excitation beam. Fluorescence is measured orthogonally using a s barrier filter to block scattered laser excitation light, and a photomultiplier tube detector output signals, which are shifted in phase from a reference signal and amplitude demodulated, are processed by phase-sensitive detection electronics to resolve signals from heterogeneous emissions and quantify decay lifetimes directly. The output signals are displayed as frequency distribution histograms and bivariate diagrams using a computer-based data acquisition system. Results have demonstrated signal phase shift, amplitude demodulation, and average measurement of fluorescence lifetimes on stained cells; a detection limit threshold of 300 to 500 fluorescein isothiocyanate (FITC); fluorescence measurement precision of 1.3% on alignment fluorospheres and 3.4% on propidium iodide (PI)-stained cells; the resolution of PI and FITC signals from cells stainedin combination with PI and FITC, based on differences in their decay lifetimes; and the ability to measure single decay nines by the two-phase, phase comparator, method.

  4. Non-Covalent Fluorescent Labeling of Hairpin DNA Probe Coupled with Hybridization Chain Reaction for Sensitive DNA Detection.

    PubMed

    Song, Luna; Zhang, Yonghua; Li, Junling; Gao, Qiang; Qi, Honglan; Zhang, Chengxiao

    2016-04-01

    An enzyme-free signal amplification-based assay for DNA detection was developed using fluorescent hairpin DNA probes coupled with hybridization chain reaction (HCR). The hairpin DNAs were designed to contain abasic sites in the stem moiety. Non-covalent labeling of the hairpin DNAs was achieved when a fluorescent ligand was bound to the abasic sites through hydrogen bonding with the orphan cytosine present on the complementary strand, accompanied by quench of ligand fluorescence. As a result, the resultant probes, the complex formed between the hairpin DNA and ligand, showed almost no fluorescence. Upon hybridization with target DNA, the probe underwent a dehybridization of the stem moiety containing an abasic site. The release of ligand from the abasic site to the solution resulted in an effective fluorescent enhancement, which can be used as a signal. Compared with a sensing system without HCR, a 20-fold increase in the sensitivity was achieved using the sensing system with HCR. The fluorescent intensity of the sensing system increased with the increase in target DNA concentration from 0.5 nM to 100 nM. A single mismatched target ss-DNA could be effectively discriminated from complementary target DNA. Genotyping of a G/C single-nucleotide polymorphism of polymerase chain reaction (PCR) products was successfully demonstrated with the sensing system. Therefore, integrating HCR strategy with non-covalent labeling of fluorescent hairpin DNA probes provides a sensitive and cost-effective DNA assay.

  5. Sensitive determination of proteins by its quenching effect on fluorescence of new terbium(III) complex.

    PubMed

    Leonenko, Inna; Aleksandrova, Daria; Yegorova, Alla; Antonovich, Valery; Karasyov, Aleksandr

    2012-01-01

    It is found that in hexamethylenetetramine (HMTA-HCl) buffer pH = 7.8, proteins can quench the fluorescence intensity of new terbium(III) complex with 6-[(1-hydroxy-3-oxo-6,7-dihydro-3H,5H-pyrido[3,2,1-ij]quinoline-2-carbonyl)-amino]-hexanoic acid (L). Based on this, a sensitive fluorimetric method for the determination of proteins is proposed. Under optimum conditions, the I0/I is in proportion to the concentration of protein in the range of 0.1-40.0 microg/mL for bovine serum albumin (BSA), 0.1-70.0 microg/mL for human serum albumin (HSA) and 0.1-40.0 microg/mL for immunoglobulin G (IgG). Their detection limits (S/N = 3) are 0.03 microg/mL. The interaction mechanism for the luminescence quenching is also studied.

  6. Red-emitting fluorescent organic light emitting diodes with low sensitivity to self-quenching

    NASA Astrophysics Data System (ADS)

    Forget, S.; Chenais, S.; Tondelier, D.; Geffroy, B.; Gozhyk, I.; Lebental, M.; Ishow, E.

    2010-09-01

    Concentration quenching is a major impediment to efficient organic light-emitting devices (OLEDs). We herein report on OLEDs based on a fluorescent amorphous red-emitting starbust triarylamine molecule [4-di(4'-tert-butylbiphenyl-4-yl)amino-4'-dicyanovinylbenzene, named FVIN], exhibiting a very small sensitivity to concentration quenching. OLEDs are fabricated with various doping levels of FVIN into Alq3, and show a remarkably stable external quantum efficiency of 1.5% for doping rates ranging from 5% up to 40%, which strongly relaxes the technological constraints on the doping accuracy. An efficiency of 1% is obtained for a pure undoped active region, along with deep red emission (x=0.6; y=0.35 in the Commission Internationale de l'Energie (CIE) coordinates). A comparison of FVIN with the archetypal 4-dicyanomethylene-2-methyl-6-(p-dimethylaminostyryl)-4H-pyran dye is presented in an identical multilayer OLED structure.

  7. An x-ray microprobe using focussing optics with a synchrotron radiation source

    SciTech Connect

    Thompson, A.C.; Underwood, J.H.; Wu, Y.; Giauque, R.D.

    1989-01-01

    An x-ray microprobe can be used to produce maps of the concentration of elements in a sample. Synchrotron radiation provides x-ray beams with enough intensity and collimation to make possible elemental images with femtogram sensitivity. The use of focussing x-ray mirrors made from synthetic multilayers with a synchrotron x-ray beam allows beam spot sizes of less than 10 /mu/m /times/ 10 /mu/m to be produced. Since minimal sample preparation is required and a vacuum environment is not necessary, there will be a wide variety of applications for such microprobes. 8 refs., 6 figs.

  8. Proton microprobe analysis of zinc in skeletal tissues. [Proton induced x-ray emission analysis

    SciTech Connect

    Doty, S B; Jones, K W; Kraner, H W; Shroy, R E; Hanson, A L

    1980-06-01

    A proton microprobe with windowless exit port was used to study zinc distributions in various types of skeletal tissues. The use of an external beam facilitated positioning of the targets for examination of particular points of interest. The proton microprobe is uniquely suited to this work since it combines high sensitivity for zinc determinations in thick samples with good spatial resolution. Measurements on rat and rabbit Achilles tendon showed a significant increase in zinc concentrations as the beam moved from the unmineralized collagen into the mineralized attachment site. Cartilage gave a similar result, with calcified cartilage having a greater zinc level than the articular surface on unmineralized epiphyseal cartilage.

  9. A sensitive solid-phase time-resolved fluorescence immunoassay apparatus

    NASA Astrophysics Data System (ADS)

    Wang, Yun-lei; Song, Ke-fei; Zhang, Wei-lai; Li, Jun-ling

    2009-07-01

    In the device, a He-Ne laser of flash frequency 1-20 Hz was adopted as exciting light source, and three key technical problems have been solved successfully in order to enhance the detecting sensitivity and measuring stability of the device for time-resolved fluorimmunoassays(TRFIA)[1]. The first one is to design optimum exciting optical system, so that the exciting light beam excite the sample most effectively. The second one is to have a project spectrum filter which can reduce the affection of the background light to the photomultiplier tube and also ensure influence of the stray light and mixed diffusion light to the sample fluorescence to the least, the sample fluorescence through the integrating sphere and come to the grating monochromator, The right wavelength will be chosed through changing the angle of incidence of the grating monochromator. The third one is to simulate the principle of sample averaging of BOXCAR averager. In the device, SCM was used as primary controller and CPLD was used as timing controller. Through the preparation process, signal-to-noise ratio(SNR) will be improved, also adjust delay time, ampling frequency and sampling number arbitrarily. By testing, the sensitivity is 10-12mol/L(substance marked by Eu3+), examination repeat is <=2.5%, examination linearity is from 10-8mol/L to 10-12mol/L, correlation coefficient is 99.98%(p<=0.01). The instrument is advanced for ultrasensitive detection of antigen and antibody , and solve the tumor, genetic variation, the virus protein detection.

  10. Boron analysis by electron microprobe using MoB4C layered synthetic crystals

    USGS Publications Warehouse

    McGee, J.J.; Slack, J.F.; Herrington, C.R.

    1991-01-01

    Preliminary electron microprobe studies of B distribution in minerals have been carried out using MoB4C-layered synthetic crystals to improve analytical sensitivity for B. Any microprobe measurements of the B contents of minerals using this crystal must include analyses for Cl to assess and correct for the interference of Cl X-rays on the BK?? peak. Microprobe analyses for B can be made routinely in tourmaline and other B-rich minerals, and minor B contents also can be determined in common rock-forming minerals. Incorporation of unusually high B contents in minerals other than borosilicates has been discovered in prograde and retrograde minerals in tourmalinites from the Broken Hill district, Australia, and may reflect high B activities produced during the metamorphism of tourmaline-rich rocks. -from Authors

  11. Thermo-optical characterization of fluorescent rhodamine B based temperature-sensitive nanosensors using a CMOS MEMS micro-hotplate☆

    PubMed Central

    Chauhan, Veeren M.; Hopper, Richard H.; Ali, Syed Z.; King, Emma M.; Udrea, Florin; Oxley, Chris H.; Aylott, Jonathan W.

    2014-01-01

    A custom designed microelectromechanical systems (MEMS) micro-hotplate, capable of operating at high temperatures (up to 700 °C), was used to thermo-optically characterize fluorescent temperature-sensitive nanosensors. The nanosensors, 550 nm in diameter, are composed of temperature-sensitive rhodamine B (RhB) fluorophore which was conjugated to an inert silica sol–gel matrix. Temperature-sensitive nanosensors were dispersed and dried across the surface of the MEMS micro-hotplate, which was mounted in the slide holder of a fluorescence confocal microscope. Through electrical control of the MEMS micro-hotplate, temperature induced changes in fluorescence intensity of the nanosensors was measured over a wide temperature range. The fluorescence response of all nanosensors dispersed across the surface of the MEMS device was found to decrease in an exponential manner by 94%, when the temperature was increased from 25 °C to 145 °C. The fluorescence response of all dispersed nanosensors across the whole surface of the MEMS device and individual nanosensors, using line profile analysis, were not statistically different (p < 0.05). The MEMS device used for this study could prove to be a reliable, low cost, low power and high temperature micro-hotplate for the thermo-optical characterisation of sub-micron sized particles. The temperature-sensitive nanosensors could find potential application in the measurement of temperature in biological and micro-electrical systems. PMID:25844025

  12. Shifts in the fluorescence lifetime of EGFP during bacterial phagocytosis measured by phase-sensitive flow cytometry

    NASA Astrophysics Data System (ADS)

    Li, Wenyan; Houston, Kevin D.; Houston, Jessica P.

    2017-01-01

    Phase-sensitive flow cytometry (PSFC) is a technique in which fluorescence excited state decay times are measured as fluorescently labeled cells rapidly transit a finely focused, frequency-modulated laser beam. With PSFC the fluorescence lifetime is taken as a cytometric parameter to differentiate intracellular events that are challenging to distinguish with standard flow cytometry. For example PSFC can report changes in protein conformation, expression, interactions, and movement, as well as differences in intracellular microenvironments. This contribution focuses on the latter case by taking PSFC measurements of macrophage cells when inoculated with enhanced green fluorescent protein (EGFP)-expressing E. coli. During progressive internalization of EGFP-E. coli, fluorescence lifetimes were acquired and compared to control groups. It was hypothesized that fluorescence lifetimes would correlate well with phagocytosis because phagosomes become acidified and the average fluorescence lifetime of EGFP is known to be affected by pH. We confirmed that average EGFP lifetimes consistently decreased (3 to 2 ns) with inoculation time. The broad significance of this work is the demonstration of how high-throughput fluorescence lifetime measurements correlate well to changes that are not easily tracked by intensity-only cytometry, which is affected by heterogeneous protein expression, cell-to-cell differences in phagosome formation, and number of bacterium engulfed.

  13. Shifts in the fluorescence lifetime of EGFP during bacterial phagocytosis measured by phase-sensitive flow cytometry

    PubMed Central

    Li, Wenyan; Houston, Kevin D.; Houston, Jessica P.

    2017-01-01

    Phase-sensitive flow cytometry (PSFC) is a technique in which fluorescence excited state decay times are measured as fluorescently labeled cells rapidly transit a finely focused, frequency-modulated laser beam. With PSFC the fluorescence lifetime is taken as a cytometric parameter to differentiate intracellular events that are challenging to distinguish with standard flow cytometry. For example PSFC can report changes in protein conformation, expression, interactions, and movement, as well as differences in intracellular microenvironments. This contribution focuses on the latter case by taking PSFC measurements of macrophage cells when inoculated with enhanced green fluorescent protein (EGFP)-expressing E. coli. During progressive internalization of EGFP-E. coli, fluorescence lifetimes were acquired and compared to control groups. It was hypothesized that fluorescence lifetimes would correlate well with phagocytosis because phagosomes become acidified and the average fluorescence lifetime of EGFP is known to be affected by pH. We confirmed that average EGFP lifetimes consistently decreased (3 to 2 ns) with inoculation time. The broad significance of this work is the demonstration of how high-throughput fluorescence lifetime measurements correlate well to changes that are not easily tracked by intensity-only cytometry, which is affected by heterogeneous protein expression, cell-to-cell differences in phagosome formation, and number of bacterium engulfed. PMID:28091553

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

    PubMed Central

    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

  15. Direct measurement of efflux in Pseudomonas aeruginosa using an environment-sensitive fluorescent dye.

    PubMed

    Iyer, Ramkumar; Erwin, Alice L

    2015-01-01

    Resistance-Nodulation-Division (RND) family pumps AcrB and MexB are the major efflux routes in Escherichia coli and Pseudomonas aeruginosa respectively. Fluorescent environment-sensitive dyes provide a means to study efflux pump function in live bacterial cells in real-time. Recently, we demonstrated the utility of this approach using the dye Nile Red to quantify AcrB-mediated efflux and measured the ability of antibiotics and other efflux pump substrates to compete with efflux of Nile Red, independent of antibacterial activity. Here, we extend this method to P. aeruginosa and describe a novel application that permits the comparison and rank-ordering of bacterial strains by their inherent efflux potential. We show that glucose and l-malate re-energize Nile Red efflux in P. aeruginosa, and we highlight differences in the glucose dependence and kinetics of efflux between P. aeruginosa and E. coli. We quantify the differences in efflux among a set of P. aeruginosa laboratory strains, which include PAO1, the hyper-sensitive strain ATCC 35151 and its parent, ATCC 12055. Efflux of Nile Red in P. aeruginosa is mediated by MexAB-OprM and is slower than in E. coli. In conclusion, we describe an efflux measurement tool for use in antibacterial drug discovery and basic research on P. aeruginosa efflux pumps.

  16. Terbium fluorescence as a sensitive, inexpensive probe for UV-induced damage in nucleic acids.

    PubMed

    El-Yazbi, Amira F; Loppnow, Glen R

    2013-07-05

    Much effort has been focused on developing methods for detecting damaged nucleic acids. However, almost all of the proposed methods consist of multi-step procedures, are limited, require expensive instruments, or suffer from a high level of interferences. In this paper, we present a novel simple, inexpensive, mix-and-read assay that is generally applicable to nucleic acid damage and uses the enhanced luminescence due to energy transfer from nucleic acids to terbium(III) (Tb(3+)). Single-stranded oligonucleotides greatly enhance the Tb(3+) emission, but duplex DNA does not. With the use of a DNA hairpin probe complementary to the oligonucleotide of interest, the Tb(3+)/hairpin probe is applied to detect ultraviolet (UV)-induced DNA damage. The hairpin probe hybridizes only with the undamaged DNA. However, the damaged DNA remains single-stranded and enhances the intrinsic fluorescence of Tb(3+), producing a detectable signal directly proportional to the amount of DNA damage. This allows the Tb(3+)/hairpin probe to be used for sensitive quantification of UV-induced DNA damage. The Tb(3+)/hairpin probe showed superior selectivity to DNA damage compared to conventional molecular beacons probes (MBs) and its sensitivity is more than 2.5 times higher than MBs with a limit of detection of 4.36±1.2 nM. In addition, this probe is easier to synthesize and more than eight times cheaper than MBs, which makes its use recommended for high-throughput, quantitative analysis of DNA damage.

  17. Fast and sensitive DNA hybridization assays using microwave-accelerated metal-enhanced fluorescence.

    PubMed

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

    2006-09-22

    A new, fast, and sensitive DNA hybridization assay platform based on microwave-accelerated metal-enhanced fluorescence (MAMEF) is presented. Thiolated oligonucleotide anchors were immobilized onto silver nanoparticles on a glass substrate. The hybridization of the complementary fluorescein-labeled DNA target with the surface-bound oligonucleotides was completed within 20 s upon heating with low-power microwaves. In addition, the signal is optically amplified, a consequence of close proximity of the fluorophore to the silvered substrate. In this proof-of-principle methodology, as low as 50 nM of a target DNA was detected, although we envisage far-lower detection limits. Control experiments, where the surface-bound oligonucleotide was omitted, were also performed to determine the extent of non-specific binding. In these studies a significantly reduced non-specific adsorption was found when using microwave heating near to silvered structures as compared to room temperature incubation. These findings suggest that MAMEF could be a most useful alternative to the DNA hybridization assays used today, especially with regard to substantially increasing both the assay rapidity and sensitivity.

  18. Data acquisition with a nuclear microprobe

    SciTech Connect

    Maggiore, C.

    1980-01-01

    Spatially resolved information from the near surfaces of materials can be obtained with a nuclear microprobe. The spatial resolution is determined by the optics of the instrument and radiation damage in the specimen. Two- and three-dimensional maps of elemental concentration may be obtained from the near surfaces of materials. Data are acquired by repeated scans of a constantly moving beam over the region of interest or by counting for a preset integrated charge at each specimen location.

  19. Biomedical application of the nuclear microprobe

    NASA Astrophysics Data System (ADS)

    Lindh, Ulf

    1987-04-01

    The Studsvik Nuclear Microprobe (SMP) has mainly been devoted to applications in the biomedical field. Its ultimate resolution is reached at 2.9×2.9 μm 2 with a proton current of 100 pA. With this performance the SMP has been used in a wide range of disciplines covering environmental hygiene, toxicology, various aspects of internal medicine and trace element physiology. Examples of recent applications in these fields are described.

  20. Preparation of antibodies and development of a sensitive immunoassay with fluorescence detection for triazine herbicides.

    PubMed

    Herranz, Sonia; Ramón-Azcón, Javier; Benito-Peña, Elena; Marazuela, María Dolores; Marco, María Pilar; Moreno-Bondi, María Cruz

    2008-07-01

    Specific polyclonal antibodies against s-triazine herbicides were obtained by preparing immunogens coupling home-synthesized haptens derivatives of simazine (6-chloro-N-ethyl-N'-ethyl-1,3,5-triazine-2,4-diamine) to lysine groups of hemocyanin from keyhole limpets and bovine serum albumin carrier proteins. Three highly sensitive rabbit antisera were obtained and evaluated with a battery of six enzyme tracers derived from triazine structures in an optimized ELISA format. The antiserum As8 and the HRP-2f tracer, which yield the best assay sensitivity for simazine (detection limit 0.11 +/- 0.02 microg L(-1), IC(50) 0.88 +/- 0.04 microg L(-1)), were applied to the development of a sensitive flow-through immunoassay for the analysis of this herbicide. The automated assay was based on a direct competitive immunosorbent assay and fluorescence detection. The optimized method presents an IC(50) value of 0.35 +/- 0.04 microg L(-1) with a detection limit of 1.3 +/- 0.9 ng L(-1) and a dynamic range from 0.010 to 7.5 microg L(-1) simazine. The generic nature of the antiserum was shown by good relative cross-reactivities with other triazines such as atrazine (420%) or propazine (130%) and a lower response to terbutylazine (6.4%) and desethyl-atrazine (2.2%). No cross-reactivity was obtained for nonrelated pesticides such as 2,4-dichlorophenoxyacetic acid or linuron and the assay could be applied as a screening method for triazine herbicides. The total analysis time was 30 min per determination and the immunosensor could be reused for more than 150 cycles without significant loss of activity. The immunosensor has been successfully applied to the direct analysis of simazine in surface water samples at the nanogram per liter level. The results obtained by comparative analysis of the immunosensor with a chromatographic procedure for triazines showed a close correspondence.

  1. Fluoroimmunoassay based on suppression of fluorescence self-quenching for ultra-sensitive detection of herbicide diuron.

    PubMed

    Sharma, Priyanka; Gandhi, Sonu; Chopra, Adity; Sekar, N; Raman Suri, C

    2010-08-31

    A highly sensitive heterogeneous fluoroimmunoassay has been developed for monitoring phenylurea herbicide diuron on the basis of suppression of fluorescence self-quenching. Specific antibody against diuron was produced and labeled with rhodamine isothiocyanate at different molar ratios and used as tracer in the developed immunoassay. The analytical sensitivity of immunoassay was enhanced by changing the microenvironment of fluorescence label with glycerin solution after the completion of immunoassay. Enhancer treatment on developed immunoassay showed improvement of fluorescence signal intensity by approximately 4-folds with higher stability compared with the signal determined without enhancer treatment of the wells. The immunoassay has a detection limit of 0.1 ng mL(-1) with good signal precision (approximately 2%) in the optimum working concentration range between 0.01 and 100 ng mL(-1) of diuron. In addition, the use of enhancer improved the stability of fluorescence signal by suppression of self-quenching of fluorescence signal. The proposed method has been applied satisfactorily for the ultra-sensitive detection of herbicide diuron in samples.

  2. A Comparison of the Capability of Sensitivity Level 3 and Sensitivity Level 4 Fluorescent Penetrants to Detect Fatigue Cracks in Aluminum

    NASA Technical Reports Server (NTRS)

    Parker, Bradford, H.

    2009-01-01

    Historically both sensitivity level 3 and sensitivity level 4 fluorescent penetrants have been used to perform NASA Standard Level inspections of aerospace hardware. In April 2008, NASA-STD-5009 established a requirement that only sensitivity level 4 penetrants were acceptable for inspections of NASA hardware. Having NASA contractors change existing processes or perform demonstration tests to certify sensitivity level 3 penetrants posed a potentially huge cost to the Agency. This study was conducted to directly compare the probability of detection sensitivity level 3 and level 4 penetrants using both Method A and Method D inspection processes. The study results strongly support the conclusion that sensitivity level 3 penetrants are acceptable for NASA Standard Level inspections

  3. Highly Sensitive Detection of S-Nitrosylated Proteins by Capillary Gel Electrophoresis with Laser Induced Fluorescence

    PubMed Central

    Wang, Siyang; Circu, Magdalena L.; Zhou, Hu; Figeys, Daniel; Aw, Tak Y.; Feng, June

    2011-01-01

    S-nitrosylated proteins are biomarkers of oxidative damage in aging and Alzheimer’s disease (AD). Here, we report a new method for detecting and quantifying nitrosylated proteins by capillary gel electrophoresis with laser induced fluorescence detection (CGE-LIF). Dylight 488 maleimide was used to specifically label thiol group (SH) after switching the S-nitrosothiol(S-NO) to SH in cysteine using the “fluorescence switch” assay. In vitro nitrosylation model-BSA subjected to S-nitrosoglutathione(GSNO) optimized the labeling reactions and characterized the response of the LIF detector. The method proves to be highly sensitive, detecting 1.3 picomolar (pM)concentration of nitrosothiols in nanograms of proteins, which is the lowest limit of detection of nitrosothiols reported to date. We further demonstrated the direct application of this method in monitoring protein nitrosylation damage in MQ mediated human colon adenocarcinoma cells. The nitrosothiol amounts in MQ treated and untreated cells are 14.8±0.2 and 10.4±0.5 pmol/mg of proteins, respectively. We also depicted nitrosylated protein electrophoretic profiles of brain cerebrum of 5-month-old AD transgenic (Tg) mice model. In Tg mice brain, 15.5±0.4 pmol of nitrosothiols/mg of proteins was quantified while wild type contained 11.7±0.3 pmol/mg proteins. The methodology is validated to quantify low levels of S-nitrosylated protein in complex protein mixtures from both physiological and pathological conditions. PMID:21820121

  4. Sensitive signal-on fluorescent sensing for copper ions based on the polyethyleneimine-capped silver nanoclusters-cysteine system.

    PubMed

    Zhang, Na; Qu, Fei; Luo, Hong Qun; Li, Nian Bing

    2013-08-12

    In this work, we present a label-free sensor for copper ions. This sensor is composed of silver nanoclusters and cysteine. The fluorescence of the silver nanoclusters was quenched by cysteine, which was recovered in the presence of copper ions. This binding of silver nanoclusters to cysteine promoted agglomeration of silver nanoclusters to yield larger non-fluorescent silver nanoparticles. The presence of copper ions resulted in the oxidation of cysteine to form a disulfide compound, leading to recovery of fluorescence of the silver nanoclusters. The fluorescence of the silver nanoclusters in the presence of cysteine increased with increasing concentration of copper ions in the range of 10-200 nM. The detection limit of this sensor for copper ions was 2.3 nM. The silver nanoclusters-cysteine sensor provides a simple, cost-effective, and sensitive platform for the detection of copper ions.

  5. A label-free fluorescent aptasensor for selective and sensitive detection of streptomycin in milk and blood serum.

    PubMed

    Taghdisi, Seyed Mohammad; Danesh, Noor Mohammad; Nameghi, Morteza Alinezhad; Ramezani, Mohammad; Abnous, Khalil

    2016-07-15

    Sensitive and fast detection of antibiotic residues in animal derived foods and blood serum is of great interest. In this study a fluorescent aptasensor was designed for selective and sensitive detection of streptomycin (STR) based on Exonuclease III (Exo III), SYBR Gold and aptamer complimentary strand. In the absence of STR, the fluorescence intensity is weak. Upon addition of STR, the aptamer binds to its target, leading to release of complementary strand from aptamer and more protection against Exo III function. Following addition of SYBR Gold, a strong fluorescence intensity is obtained. This aptasensor showed a high selectivity toward STR with a limit of detection (LOD) as low as 54.5 nM. The validity of the procedure and applicability of the aptasensor were successfully assessed by detection of STR in a spiked milk and blood serum without interference from the sample matrix.

  6. Fluorescent trimethyl-substituted naphthyridine as a label-free signal reporter for one-step and highly sensitive fluorescent detection of DNA in serum samples.

    PubMed

    Wang, Jiamian; Wang, Xiuyun; Wu, Shuo; Che, Ruping; Luo, Pinchen; Meng, Changgong

    2017-01-15

    A facile label-free sensing method is developed for the one-step and highly sensitive fluorescent detection of DNA, which couples the specific C-C mismatch bonding and fluorescent quenching property of a trimethyl-substituted naphthyridine dye (ATMND) with the exonuclease III (Exo III) assisted cascade target recycling amplification strategy. In the absence of target DNA, the DNA hairpin probe with a C-C mismatch in the stem and more than 4 bases overhung at the 3' terminus could entrap and quench the fluorescence of ATMND and resist the digestion of Exo III, thus showing a low fluorescence background. In the presence of the target, however, the hybridization event between the two protruding segments and the target triggers the digestion reaction of Exo III, recycles the initial target, and simultaneously releases both the secondary target analogue and the ATMND caged in the stem. The released initial and secondary targets take part in another cycle of digestion, thus leading to the release of a huge amount of free ATMND for signal transducing. Based on the fluorescence recovery, the as-proposed label-free fluorescent sensing strategy shows very good analytical performances towards DNA detection, such as a wide linear range from 10pM to 1μM, a low limit of detection of 6pM, good selectivity, and a facile one-step operation at room temperature. Practical sample analysis in serum samples indicates the method has good precision and accuracy, which may thus have application potentials for point-of-care screening of DNA in complex clinical and environmental samples.

  7. Enhanced Sensitivity for Detection of HIV-1 p24 Antigen by a Novel Nuclease-Linked Fluorescence Oligonucleotide Assay

    PubMed Central

    Fan, Peihu; Li, Xiaojun; Su, Weiheng; Kong, Wei; Kong, Xianggui; Wang, Zhenxin; Wang, Youchun; Jiang, Chunlai; Gao, Feng

    2015-01-01

    The relatively high detection limit of the Enzyme-linked immunosorbent assay (ELISA) prevents its application for detection of low concentrations of antigens. To increase the sensitivity for detection of HIV-1 p24 antigen, we developed a highly sensitive nuclease-linked fluorescence oligonucleotide assay (NLFOA). Two major improvements were incorporated in NLFOA to amplify antibody-antigen interaction signals and reduce the signal/noise ratio; a large number of nuclease molecules coupled to the gold nanoparticle/streptavidin complex and fluorescent signals generated from fluorescent-labeled oligonucleotides by the nuclease. The detection limit of p24 by NLFOA was 1 pg/mL, which was 10-fold more sensitive than the conventional ELISA (10 pg/mL). The specificity was 100% and the coefficient of variation (CV) was 7.8% at low p24 concentration (1.5 pg/mL) with various concentrations of spiked p24 in HIV-1 negative sera. Thus, NLFOA is highly sensitive, specific, reproducible and user-friendly. The more sensitive detection of low p24 concentrations in HIV-1-infected individuals by NLFOA could allow detection of HIV-1 infections that are missed by the conventional ELISA at the window period during acute infection to further reduce the risk for HIV-1 infection due to the undetected HIV-1 in the blood products. Moreover, NLFOA can be easily applied to more sensitive detection of other antigens. PMID:25915630

  8. Boronic Acid: A Bio-Inspired Strategy To Increase the Sensitivity and Selectivity of Fluorescent NADH Probe.

    PubMed

    Wang, Lu; Zhang, Jingye; Kim, Beomsue; Peng, Juanjuan; Berry, Stuart N; Ni, Yong; Su, Dongdong; Lee, Jungyeol; Yuan, Lin; Chang, Young-Tae

    2016-08-24

    Fluorescent probes have emerged as an essential tool in the molecular recognition events in biological systems; however, due to the complex structures of certain biomolecules, it remains a challenge to design small-molecule fluorescent probes with high sensitivity and selectivity. Inspired by the enzyme-catalyzed reaction between biomolecule and probe, we present a novel combination-reaction two-step sensing strategy to improve sensitivity and selectivity. Based on this strategy, we successfully prepared a turn-on fluorescent reduced nicotinamide adenine dinucleotide (NADH) probe, in which boronic acid was introduced to bind with NADH and subsequently accelerate the sensing process. This probe shows remarkably improved sensitivity (detection limit: 0.084 μM) and selectivity to NADH in the absence of any enzymes. In order to improve the practicality, the boronic acid was further modified to change the measurement conditions from alkalescent (pH 9.5) to physiological environment (pH 7.4). Utilizing these probes, we not only accurately quantified the NADH weight in a health care product but also evaluated intracellular NADH levels in live cell imaging. Thus, these bio-inspired fluorescent probes offer excellent tools for elucidating the roles of NADH in biological systems as well as a practical strategy to develop future sensitive and selective probes for complicated biomolecules.

  9. An Iron Microprobe Study of Be-B Isotope Systematic in Melilite-Rich CAIs Based on Newly Determined Be/B Relative Sensitivity Factors for Melilitic Glass Standards

    NASA Astrophysics Data System (ADS)

    Fukuda, K.; Fujiya, W.; Hiyagon, H.; Sugiura, N.; Takahata, N.; Sano, Y.

    2016-08-01

    We report relative sensitivity factors for melilitic glasses and Be-B systematics of melilite-rich CAIs in CO, CH and ungrouped C chondrites. The variable 10Be/9Be ratios observed in this study supports 10Be production by local irradiation processes.

  10. Integrated platform with magnetic purification and rolling circular amplification for sensitive fluorescent detection of ochratoxin A.

    PubMed

    Yao, Li; Chen, Yinji; Teng, Jun; Zheng, Wanli; Wu, Jingjing; Adeloju, Samuel B; Pan, Daodong; Chen, Wei

    2015-12-15

    In this article, we report the detection of ochratoxin A (OTA) with excellent sensitivity with the two-aspect signal amplification treatments. Combining the unique property of magnetic nanoparticles and the high efficiency of the in vitro amplification of rolling circular amplification (RCA), the competitive sensing protocol for ultrasensitive detection of OTA was achieved in about 80 min. The excellent magnetic separation treatment could effectively avoid the interference of background fluorescent noise in the sensing system while the RCA could tremendously increase the hybridization sequence for the quantum dot labeled probes and further increase the sensing response signal. Afterwards, two factors affecting the final detection limit, concentration of RCA components and RCA reaction time, were all systematically optimized for the best sensing performance. The response of the optimized protocol for OTA detection is highly linear over the wider range from 10(-3) to 10 ppb, which is 3 orders improvement in sensing range, and the limit of detection is calculated to be as low as 0.13 ppt, which is 10,000 folds improvement compared with the traditional methods. More importantly, given the selected aptamer, this universal signal amplification protocol could be widely applied to other fields by just change the recognition sequence of the aptamer.

  11. Sensitive detection of Ochratoxin A in food and drinks using metal-enhanced fluorescence.

    PubMed

    Todescato, Francesco; Antognoli, Agnese; Meneghello, Anna; Cretaio, Erica; Signorini, Raffaella; Bozio, Renato

    2014-07-15

    Easy, sensitive, rapid and low cost ochratoxin biosensors are strongly demanded in food analysis since Ochratoxin A (OTA) is a widely diffused food contaminant, highly detrimental for human health. In this work, a novel plasmonic based optical biosensor prototype for ochratoxin A is described. It exploits the metal-enhanced fluorescence phenomenon due to the silver film over nanosphere plasmonic substrate. Since ochratoxin A could be present in different food commodities, sensor performances have been tested on three different matrices (dried milk, juices, and wheat mix). Firstly, a common OTA extraction solvent and a labeling and detection protocol were defined for the analyzed matrices. Then, the efficiency of the Ag-FON surfaces in signal amplification for the detection of low ochratoxin A concentrations was defined. Using samples spiked with OTA-AF 647 or with unlabeled OTA we were able to detect the mycotoxin at concentrations lower than E.U. specifications of 0.5 μg/kg in wheat, milk and apple juice. The test performances are comparable to those of ELISA kits but the platform presented here, once optimized, present some perspective advantages, such as: low cost and time consuming, versatility of the protocol for the investigation of different matrices, employment also in non-qualified laboratories, small dimensions that allow its integration in a compact device for OTA on-site detection.

  12. Simple and sensitive determination of five quinolones in food by liquid chromatography with fluorescence detection.

    PubMed

    Ramos, Macarena; Aranda, Angela; Garcia, Elena; Reuvers, Thea; Hooghuis, Henny

    2003-06-15

    A simple and sensitive high-performance liquid chromatographic (HPLC) method has been developed for the determination of five different quinolones: enrofloxacin, ciprofloxacin, sarafloxacin, oxolinic acid and flumequine in pork and salmon muscle. The method includes one extraction and clean-up step for the five quinolones together which are detected in two separated HPLC runs by means of their fluorescence. The proposed analytical method involves homogenizing of the tissue sample with 0.05 M phosphate buffer, pH 7.4 and clean-up by Discovery DS-18 cartridges. For chromatographic separation a Symmetry C(18) column is used in two different runs: (1) ciprofloxacin, enrofloxacin and sarafloxacin with acetonitrile-0.02 M phosphate buffer pH 3.0 (18:82) as mobile phase and the detector at excitation wavelength: 280 nm and emission wavelength 450 nm; and (2) oxolinic acid and flumequine with acetonitrile-0.02 M phosphate buffer pH 3.0 (34:66) as mobile phase and excitation wavelength: 312 nm and emission wavelength: 366 nm. Detection limit was as low as 5 ng g(-1), except for sarafloxacin which had a limit of 10 ng g(-1). Standard curves using blank muscle tissues spiked at different levels showed a good linear correlation coefficient, r(2) higher than 0.999 for all quinolones.

  13. A two-photon laser induced fluorescence diagnostic with improved sensitivity, localization, and measurement rate

    NASA Astrophysics Data System (ADS)

    Elliott, Drew; Scime, Earl; Short, Zachary

    2016-10-01

    A two-photon absorption laser induced fluorescence diagnostic has been developed for measuring neutrals in fusion plasmas. Implementation of this diagnostic on the HIT-SI3 spheromak has demonstrated the sensitivity of the diagnostic and shown that measurements taken over several plasma pulses are possible. These measurements yielded an unexpected loss of signal when complex collection optics were utilized. Simulations show that this loss of signal can be explained by chromatic aberrations caused by the disparate Kr and D emission. This loss of signal has been addressed with the development of a new calibration scheme involving xenon gas. The Xe calibration scheme emission occurs at 656.00 nm while the deuterium emission is 656.09 nm. This nearly identical emission allows for advanced optical techniques such as confocal collection/injection and spatial filtering to be employed without loss of signal. Spatial filtering has been demonstrated to decrease noise while improving measurement localization, while confocal collection/injection allows for probing and measuring to occur through one viewport. The Xe scheme also allows for a Doppler-free hydrogen measurement. Doppler-free measurements eliminate the need to scan the laser spectrally thus greatly increasing the rate of measurement.

  14. Sensitive determination of ranitidine in rabbit plasma by HPLC with fluorescence detection.

    PubMed

    Khedr, Alaa

    2008-02-01

    A sensitive high-performance liquid chromatographic method for determination of ranitidine (RAN) in rabbit plasma is described. The method is based on liquid-liquid extraction, labeling with dansyl chloride and monitoring with fluorescence detector at 338nm (ex)/523nm (em). Plasma samples were extracted with diethyl ether alkalinized with 1M sodium hydroxide. Ephedrine HCl (EPH-HCl) was used as internal standard. Both, RAN and EPH were completely derivatized after heating at 60 degrees C for 10min in sodium bicarbonate solution (pH 9.5). The derivatized samples were analyzed by HPLC using Agilent Zorbax Extended C18 column (150mmx4.6mm i.d.) and mobile phase consists of 48% acetonitrile and 52% sodium acetate solution (0.02M, pH 4.6). The linearity of the method was in the range of 0.025-10microg/ml. The limits of detection (LOD) and quantification (LOQ) were 7.5+/-0.18 and 22.5+/-0.12ng/ml, respectively. Ranitidine recovery was 97.5+/-1.1% (n=6; R.S.D.=1.8%). The method was applied on plasma collected from rabbits at different time intervals after oral administration of 5mg/kg ranitidine HCl.

  15. Sensitivity of protein array deposition using continuous flow printing for fluorescent microarray applications - biomed 2013.

    PubMed

    Romanov, Valentin; Miles, Adam; Gale, Bruce; Eckman, Josh; Brooks, Benjamin

    2013-01-01

    The promise of antibody and protein microarrays to revolutionize disease diagnostics has failed to live up to the hype primarily due to the problems associated with the printing of the antibodies and/or proteins onto the detection surface. The current standard in printing proteins is pin printing. An alternative to the pin printer is the continuous-flow microspotter (CFM), a protein printer that uses microfluidic flow to print down the proteins. The advantages of the CFM include consistent spot morphology, spot-to-spot uniformity and enhanced surface concentration. Further, the CFM is effective at capturing proteins and antibodies from either dilute or complex (e.g. blood or tissue) samples. In this study, the sensitivity of CFM printing Cy3 and Cy5 fluorescently labeled proteins was determined. Values were obtained at low concentrations tens of ng/mL with low coefficients of variation. Thus, the CFM can effectively print and quantify proteins and antibodies from low concentration and complex buffered samples.

  16. Behavior of plant plasma membranes under hydrostatic pressure as monitored by fluorescent environment-sensitive probes.

    PubMed

    Roche, Yann; Klymchenko, Andrey S; Gerbeau-Pissot, Patricia; Gervais, Patrick; Mély, Yves; Simon-Plas, Françoise; Perrier-Cornet, Jean-Marie

    2010-08-01

    We monitored the behavior of plasma membrane (PM) isolated from tobacco cells (BY-2) under hydrostatic pressures up to 3.5kbar at 30 degrees C, by steady-state fluorescence spectroscopy using the newly introduced environment-sensitive probe F2N12S and also Laurdan and di-4-ANEPPDHQ. The consequences of sterol depletion by methyl-beta-cyclodextrin were also studied. We found that application of hydrostatic pressure led to a marked decrease of hydration as probed by F2N12S and to an increase of the generalized polarization excitation (GPex) of Laurdan. We observed that the hydration effect of sterol depletion was maximal between 1 and 1.5 kbar but was much less important at higher pressures (above 2 kbar) where both parameters reached a plateau value. The presence of a highly dehydrated gel state, insensitive to the sterol content, was thus proposed above 2.5 kbar. However, the F2N12S polarity parameter and the di-4-ANEPPDHQ intensity ratio showed strong effect on sterol depletion, even at very high pressures (2.5-3.5 kbar), and supported the ability of sterols to modify the electrostatic properties of membrane, notably its dipole potential, in a highly dehydrated gel phase. We thus suggested that BY-2 PM undergoes a complex phase behavior in response to the hydrostatic pressure and we also emphasized the role of phytosterols to regulate the effects of high hydrostatic pressure on plant PM.

  17. A reusable and sensitive biosensor for total mercury in canned fish based on fluorescence polarization.

    PubMed

    Shen, Tongfei; Yue, Qiaoli; Jiang, Xiuxiu; Wang, Lei; Xu, Shuling; Li, Haibo; Gu, Xiaohong; Zhang, Shuqiu; Liu, Jifeng

    2013-12-15

    In this work, we developed a sensitive and selective sensor technique for total mercury (Hg) detection in canned fish samples based on the fluorescence polarization (FP) method. The detection principle was that ssDNA containing thymine (T) bases was modified on magnetic nanoparticles (MNPs), which were used as enhancement probe. In the presence of Hg(2+), the ssDNA on MNPs can hybridize with the fluorophore labeled aptamer owing to the specific interaction between T bases and Hg(2+). The formation of thymine-Hg(2+)-thymine (T-Hg(2+)-T) complexes leads to the molar mass increase of fluorophore molecules, resulting in the enhancement of FP signal. The increase of FP was in a good linearity with the concentration of Hg(2+) in range of 2.0 nM-1.0 mM and the limit of detection was 0.49 nM (3.29 SB/m, according to the recent recommendation of IUPAC). Moreover, the proposed biosensor can be reused for 6 cycling times and was successfully applied in monitoring Hg(2+) in real samples.

  18. Imaging neuronal seal resistance on silicon chip using fluorescent voltage-sensitive dye.

    PubMed

    Braun, Dieter; Fromherz, Peter

    2004-08-01

    The electrical sheet resistance between living cells grown on planar electronic contacts of semiconductors or metals is a crucial parameter for bioelectronic devices. It determines the strength of electrical signal transduction from cells to chips and from chips to cells. We measured the sheet resistance by applying AC voltage to oxidized silicon chips and by imaging the voltage change across the attached cell membrane with a fluorescent voltage-sensitive dye. The phase map of voltage change was fitted with a planar core-coat conductor model using the sheet resistance as a free parameter. For nerve cells from rat brain on polylysine as well as for HEK293 cells and MDCK cells on fibronectin we find a similar sheet resistance of 10 MOmega. Taking into account the independently measured distance of 50 nm between chip and membrane for these cells, we obtain a specific resistance of 50 Omegacm that is indistinguishable from bulk electrolyte. On the other hand, the sheet resistance for erythrocytes on polylysine is far higher, at approximately 1.5 GOmega. Considering the distance of 10 nm, the specific resistance in the narrow cleft is enhanced to 1500 Omegacm. We find this novel optical method to be a convenient tool to optimize the interface between cells and chips for bioelectronic devices.

  19. Molecular engineering of a fluorescent bioprobe for sensitive and selective detection of amphibole asbestos.

    PubMed

    Ishida, Takenori; Alexandrov, Maxym; Nishimura, Tomoki; Hirota, Ryuichi; Ikeda, Takeshi; Kuroda, Akio

    2013-01-01

    Fluorescence microscopy-based affinity assay could enable highly sensitive and selective detection of airborne asbestos, an inorganic environmental pollutant that can cause mesothelioma and lung cancer. We have selected an Escherichia coli histone-like nucleoid structuring protein, H-NS, as a promising candidate for an amphibole asbestos bioprobe. H-NS has high affinity to amphibole asbestos, but also binds to an increasingly common asbestos substitute, wollastonite. To develop a highly specific Bioprobe for amphibole asbestos, we first identified a specific but low-affinity amosite-binding sequence by slicing H-NS into several fragments. Second, we constructed a streptavidin tetramer complex displaying four amosite-binding fragments, resulting in the 250-fold increase in the probe affinity as compared to the single fragment. The tetramer probe had sufficient affinity and specificity for detecting all the five types of asbestos in the amphibole group, and could be used to distinguish them from wollastonite. In order to clarify the binding mechanism and identify the amino acid residues contributing to the probe's affinity to amosite fibers, we constructed a number of shorter and substituted peptides. We found that the probable binding mechanism is electrostatic interaction, with positively charged side chains of lysine residues being primarily responsible for the probe's affinity to asbestos.

  20. Structural and permeability sensitivity of cells to low intensity ultrasound: Infrared and fluorescence evidence in vitro.

    PubMed

    Domenici, Fabio; Giliberti, Claudia; Bedini, Angelico; Palomba, Raffaele; Udroiu, Ion; Di Giambattista, Lucia; Pozzi, Deleana; Morrone, Stefania; Bordi, Federico; Congiu Castellano, Agostina

    2014-04-01

    This work is focused on the in vitro study of the effects induced by medical ultrasound (US) in murine fibroblast cells (NIH-3T3) at a low-intensity of exposure (spatial peak temporal average intensity Ita<0.1Wcm(-2)). Conventional 1MHz and 3MHz US devices of therapeutic relevance were employed with varying intensity and exposure time parameters. In this framework, upon cells exposure to US, structural changes at the molecular level were evaluated by infrared spectroscopy; alterations in plasma membrane permeability were monitored in terms of uptake efficiency of small cell-impermeable model drug molecules, as measured by fluorescence microscopy and flow cytometry. The results were related to the cell viability and combined with the statistical PCA analysis, confirming that NIH-3T3 cells are sensitive to therapeutic US, mainly at 1MHz, with time-dependent increases in both efficiency of uptake, recovery of wild-type membrane permeability, and the size of molecules entering 3T3. On the contrary, the exposures from US equipment at 3MHz show uptakes comparable with untreated samples.

  1. Sensitive arginine sensing based on inner filter effect of Au nanoparticles on the fluorescence of CdTe quantum dots

    NASA Astrophysics Data System (ADS)

    Liu, Haijian; Li, Ming; Jiang, Linye; Shen, Feng; Hu, Yufeng; Ren, Xueqin

    2017-02-01

    Arginine plays an important role in many biological functions, whose detection is very significant. Herein, a sensitive, simple and cost-effective fluorescent method for the detection of arginine has been developed based on the inner filter effect (IFE) of citrate-stabilized gold nanoparticles (AuNPs) on the fluorescence of thioglycolic acid-capped CdTe quantum dots (QDs). When citrate-stabilized AuNPs were mixed with thioglycolic acid-capped CdTe QDs, the fluorescence of CdTe QDs was significantly quenched by AuNPs via the IFE. With the presence of arginine, arginine could induce the aggregation and corresponding absorption spectra change of AuNPs, which then IFE-decreased fluorescence could gradually recover with increasing amounts of arginine, achieving fluorescence "turn on" sensing for arginine. The detection mechanism is clearly illustrated and various experimental conditions were also optimized. Under the optimum conditions, a decent linear relationship was obtained in the range from 16 to 121 μg L- 1 and the limit of detection was 5.6 μg L- 1. And satisfactory results were achieved in arginine analysis using arginine injection, compound amino acid injection, even blood plasma as samples. Therefore, the present assay showed various merits, such as simplicity, low cost, high sensitivity and selectivity, making it promising for sensing arginine in biological samples.

  2. A Validated High-Throughput Fluorometric Method for Determination of Omeprazole in Quality Control Laboratory via Charge Transfer Sensitized Fluorescence.

    PubMed

    Mahmoud, Ashraf M; Ahmed, Sameh A

    2016-03-01

    A high-throughput 96-microwell plate fluorometric method was developed and validated to determine omeprazole (OMZ) in its dosage forms. The method was based on the charge-transfer (CT) sensitized fluorescence reaction of OMZ with 2, 3-dichloro-5, 6-dicyano-1, 4-benzoquinone (DDQ). This fluorescence reaction provided a new approach for simple, sensitive and selective determinations of OMZ in pharmaceutical preparations. In the present method, the fluorescence reaction was carried out in 96-microwell plates as reaction vessels in order to increase the automation of the methodology and the efficiency of its use in quality control laboratories. All factors affecting the fluorescence reaction were carefully studied and the conditions were optimized. The stoichiometry of the fluorescence reaction between OMZ and DDQ was determined and the reaction mechanism was suggested. Under the optimum conditions, the linear range was 100-6000 ng/ml with the lowest LOD of 33 ng/ml. Analytical performance of the proposed assay, in terms of accuracy and precision, was statistically validated and the results were satisfactory; RSD was <2.6 % and the accuracy was 98.6-101.6 %. The method was successfully applied to the analysis of OMZ in its dosage forms; the recovery values were 98.26-99.60 ± 0.95-2.22 %. The developed methodology may provide a safer, automated and economic tool for the analysis of OMZ in quality control laboratories.

  3. Ag Nanoparticles-enhanced Fluorescence of Terbium-Deferasirox Complexes for the Highly Sensitive Determination of Deferasirox.

    PubMed

    Abolhasani, Jafar; Naderali, Roza; Hassanzadeh, Javad

    2016-01-01

    We describe the effect of different sized gold and silver nanoparticles on the terbium sensitized fluorescence of deferasirox. It is indicated that silver nanostructures, especially 18 nm Ag nanoparticles (AgNPs), have a remarkable amplifying effect compared to Au nanoparticles. Based on this observation, a highly sensitive and selective method was developed for the determination of deferasirox. Effects of various parameters like AgNPs and Tb(3+) concentration and pH of media were investigated. Under the optimal conditions, a calibration curve was plotted as the fluorescence intensities versus the concentration of deferasirox in the range of 0.1 to 200 nmol L(-1), and detection limit of 0.03 nmol L(-1) was obtained. The method has good linearity, recovery, reproducibility and sensitivity, and was satisfactorily applied for the determination of deferasirox in urine and pharmaceutical samples.

  4. Graphitic Carbon Nitride Nanosheets-Based Ratiometric Fluorescent Probe for Highly Sensitive Detection of H2O2 and Glucose.

    PubMed

    Liu, Jin-Wen; Luo, Ying; Wang, Yu-Min; Duan, Lu-Ying; Jiang, Jian-Hui; Yu, Ru-Qin

    2016-12-14

    Graphitic carbon nitride (g-C3N4) nanosheets, an emerging graphene-like carbon-based nanomaterial with high fluorescence and large specific surface areas, hold great potential for biosensor applications. Current g-C3N4 nanosheets based fluorescent biosensors majorly rely on single fluorescent intensity reading through fluorescence quenching interactions between the nanosheets and metal ions. Here we report for the first time the development of a novel g-C3N4 nanosheets-based ratiometric fluorescence sensing strategy for highly sensitive detection of H2O2 and glucose. With o-phenylenediamine (OPD) oxidized by H2O2 in the presence of horseradish peroxidase (HRP), the oxidization product can assemble on the g-C3N4 nanosheets through hydrogen bonding and π-π stacking, which effectively quenches the fluorescence of g-C3N4 while delivering a new emission peak. The ratiometric signal variations enable robust and sensitive detection of H2O2. On the basis of the glucose converting into H2O2 through the catalysis of glucose oxidase, the g-C3N4-based ratiometric fluorescence sensing platform is also exploited for glucose assay. The developed strategy is demonstrated to give a detection limit of 50 nM for H2O2 and 0.4 μM for glucose, at the same time, it has been successfully used for glucose levels detection in human serum. This strategy may provide a cost-efficient, robust, and high-throughput platform for detecting various species involving H2O2-generation reactions for biomedical applications.

  5. Folic acid functionalized silver nanoparticles with sensitivity and selectivity colorimetric and fluorescent detection for Hg2+ and efficient catalysis

    NASA Astrophysics Data System (ADS)

    Su, Dongyue; Yang, Xin; Xia, Qingdong; Zhang, Qi; Chai, Fang; Wang, Chungang; Qu, Fengyu

    2014-09-01

    In this research, folic acid functionalized silver nanoparticles (FA-AgNPs) were selected as a colorimetric and a ‘turn on’ fluorescent sensor for detecting Hg2+. After being added into Hg2+, AgNPs can emit stable fluorescence at 440 nm when the excitation wavelength is selected at 275 nm. The absorbance and fluorescence of the FA-AgNPs could reflect the concentration of the Hg2+ ions. Thus, we developed a simple, sensitive analytical method to detect Hg2+ based on the colorimetric and fluorescence enhancement of FA-AgNPs. The sensor exhibits two linear response ranges between absorbance and fluorescence intensity with Hg2+ concentration, respectively. Meanwhile, a detection limit of 1 nM is estimated based on the linear relationship between responses with a concentration of Hg2+. The high specificity of Hg2+ with FA-AgNPs interactions provided the excellent selectivity towards detecting Hg2+ over other metal ions (Pb2+, Mg2+, Zn2+, Ni2+, Cu2+, Co2+, Ca2+, Mn2+, Fe2+, Cd2+, Ba2+, Cr6+ and Cr3+). This will provide a simple, effective and multifunctional colorimetric and fluorescent sensor for on-site and real-time Hg2+ ion detection. The proposed method can be applied to the analysis of trace Hg2+ in lake water. Additionally, the FA-AgNPs can be used as efficient catalyst for the reduction of 4-nitrophenol and potassium hexacyanoferrate (III).

  6. Label-free silicon quantum dots as fluorescent probe for selective and sensitive detection of copper ions.

    PubMed

    Zhao, Jiangna; Deng, Jianhui; Yi, Yinhui; Li, Haitai; Zhang, Youyu; Yao, Shouzhuo

    2014-07-01

    In this work, label-free silicon quantum dots (SiQDs) were used as a novel fluorescence probe for the sensitive and selective detection of Cu(2+). The fluorescence of the SiQDs was effectively quenched by H2O2 from the reaction of ascorbic acid with O2, and hydroxyl radicals from Fenton reaction between H2O2 and Cu(+). The fluorescence intensity of SiQDs was quenched about 25% in 15 min after the addition of H2O2 (1mM). While the SiQDs was incubated with AA (1mM) and Cu(2+) (1 µM) under the same conditions, the fluorescence intensity of SiQDs decreased about 55%. Obviously, the recycling of Cu(2+) in the test system may lead to a dramatical decrease in the fluorescence of SiQDs. Under the optimized experimental conditions, the rate of fluorescence quenching of SiQDs was linearly dependent on the Cu(2+) concentration ranging from 25 to 600 nM with the limit of detection as low as 8 nM, which was much lower than that of existing methods. Moreover, the probe was successfully applied to the determination of Cu(2+) in different environmental water samples and human hair.

  7. Sulfur-doped graphene quantum dots as a novel fluorescent probe for highly selective and sensitive detection of Fe(3+).

    PubMed

    Li, Shuhua; Li, Yunchao; Cao, Jun; Zhu, Jia; Fan, Louzhen; Li, Xiaohong

    2014-10-21

    Sulfur-doped graphene quantum dots (S-GQDs) with stable blue-green fluorescence were synthesized by one-step electrolysis of graphite in sodium p-toluenesulfonate aqueous solution. Compared with GQDs, the S-GQDs drastically improved the electronic properties and surface chemical reactivities, which exhibited a sensitive response to Fe(3+). Therefore, the S-GQDs were used as an efficient fluorescent probe for highly selective detection of Fe(3+). Upon increasing of Fe(3+) concentration ranging from 0.01 to 0.70 μM, the fluorescence intensity of S-GQDs gradually decreased and reached a plateau at 0.90 μM. The difference in the fluorescence intensity of S-GQDs before and after adding Fe(3+) was proportional to the concentration of Fe(3+), and the calibration curve displayed linear regions over the range of 0-0.70 μM. The detection limit was 4.2 nM. Finally, this novel fluorescent probe was successfully applied to the direct analysis of Fe(3+) in human serum, which presents potential applications in clinical diagnosis and may open a new way to the design of effective fluorescence probes for other biologically related targets.

  8. A carbon dot-based "off-on" fluorescent probe for highly selective and sensitive detection of phytic acid.

    PubMed

    Gao, Zhao; Wang, Libing; Su, Rongxin; Huang, Renliang; Qi, Wei; He, Zhimin

    2015-08-15

    We herein report a facile, one-step pyrolysis synthesis of photoluminescent carbon dots (CDs) using citric acid as the carbon source and lysine as the surface passivation reagent. The as-prepared CDs show narrow size distribution, excellent blue fluorescence and good photo-stability and water dispersivity. The fluorescence of the CDs was found to be effectively quenched by ferric (Fe(III)) ions with high selectivity via a photo-induced electron transfer (PET) process. Upon addition of phytic acid (PA) to the CDs/Fe(III) complex dispersion, the fluorescence of the CDs was significantly recovered, arising from the release of Fe(III) ions from the CDs/Fe(III) complex because PA has a higher affinity for Fe(III) ions compared to CDs. Furthermore, we developed an "off-on" fluorescence assay method for the detection of phytic acid using CDs/Fe(III) as a fluorescent probe. This probe enables the selective detection of PA with a linear range of 0.68-18.69 μM and a limit of detection (signal-to-noise ratio is 3) of 0.36 μM. The assay method demonstrates high selectivity, repeatability, stability and recovery ratio in the detection of the standard and real PA samples. We believe that the facile operation, low-cost, high sensitivity and selectivity render this CD-based "off-on" fluorescent probe an ideal sensing platform for the detection of PA.

  9. Intravital excitation increases detection sensitivity for pulmonary tuberculosis by whole-body imaging with β-lactamase reporter enzyme fluorescence.

    PubMed

    Nooshabadi, Fatemeh; Yang, Hee-Jeong; Cheng, Yunfeng; Durkee, Madeleine S; Xie, Hexin; Rao, Jianghong; Cirillo, Jeffrey D; Maitland, Kristen C

    2016-10-18

    Tuberculosis is a pulmonary disease with an especially high mortality rate in immuno-compromised populations, specifically children and HIV positive individuals. The causative agent, Mycobacterium tuberculosis (Mtb), is a very slow growing and difficult organism to work with, making both diagnosis and development of effective treatments cumbersome. We utilize a fiber-optic fluorescence microendoscope integrated with a whole-body imaging system for in vivo Mtb detection. The system exploits an endogenous enzyme of Mtb (β-lactamase, or BlaC) using a BlaC-specific NIR fluorogenic substrate. In the presence of BlaC, this substrate is cleaved and becomes fluorescent. Using intravital illumination of the lung to excite this probe, sensitivity of the optical system increases over trans- and epi-illumination methods of whole-body fluorescence imaging. We demonstrate that integration of these imaging technologies with BlaC-specific fluorescent reporter probe improves the level of detection to ∼100 colony forming units, a 100× increase in sensitivity in comparison to epi-illumination and a 10× increase in sensitivity in comparison to previous work in intravital excitation of tdTomato-expressing Mtb. This lower detection threshold enables the study of early stage bacterial infections with clinical strains of Mtb and longitudinal studies of disease pathogenesis and therapeutic efficacy with multiple time points in a single animal.

  10. Time-Resolved Fluorescent Immunochromatography of Aflatoxin B1 in Soybean Sauce: A Rapid and Sensitive Quantitative Analysis

    PubMed Central

    Wang, Du; Zhang, Zhaowei; Li, Peiwu; Zhang, Qi; Zhang, Wen

    2016-01-01

    Rapid and quantitative sensing of aflatoxin B1 with high sensitivity and specificity has drawn increased attention of studies investigating soybean sauce. A sensitive and rapid quantitative immunochromatographic sensing method was developed for the detection of aflatoxin B1 based on time-resolved fluorescence. It combines the advantages of time-resolved fluorescent sensing and immunochromatography. The dynamic range of a competitive and portable immunoassay was 0.3–10.0 µg·kg−1, with a limit of detection (LOD) of 0.1 µg·kg−1 and recoveries of 87.2%–114.3%, within 10 min. The results showed good correlation (R2 > 0.99) between time-resolved fluorescent immunochromatographic strip test and high performance liquid chromatography (HPLC). Soybean sauce samples analyzed using time-resolved fluorescent immunochromatographic strip test revealed that 64.2% of samples contained aflatoxin B1 at levels ranging from 0.31 to 12.5 µg·kg−1. The strip test is a rapid, sensitive, quantitative, and cost-effective on-site screening technique in food safety analysis. PMID:27428975

  11. Metal-enhanced fluorescent dye-doped silica nanoparticles and magnetic separation: A sensitive platform for one-step fluorescence detection of prostate specific antigen.

    PubMed

    Xu, Dang-Dang; Deng, Yun-Liang; Li, Cheng-Yu; Lin, Yi; Tang, Hong-Wu

    2017-01-15

    The world health organization figures show prostate cancer in developed countries has been the second primary cause of cancer mortality following lung cancer for the men. So, early and sensitive diagnosis of cancer is very important before it spreads out to the other organs of the body. It is well-known that prostate-specific antigen (PSA) is the most specific and efficient tumor marker for the diagnosis of prostate cancer. Herein, we successfully fabricated core-shell composite fluorescent nanoparticle Ag@SiO2@SiO2-RuBpy which provide a photoluminescence enhancement of up to ~3-fold when the separation distance between the surface of silver core and the center of the third RuBpy doped silica shell is about 10nm. These core-shell MEF-capable nanoparticles have obvious advantages. The interaction between the doped RuBpy molecules in the outer silica layer and the silver core, greatly improves the excitation efficiency and enhances the fluorescence intensity. Importantly, the presence of silica can reduce the self-quenching of RuBpy, which makes larger amounts of RuBpy incorporated into the silica shell. In addition, the shell protects the RuBpy against collisional quenching and irreversible photodegradation and provides abundant hydroxyl for easy conjugation. After that a highly sensitive, specific and reliable strategy based on metal-enhanced fluorescence and magnetic separation was applied for the detection of PSA in both buffer and serum. The process could be rapidly accomplished, in which the immunomagnetic nanospheres (IMNs) and immunofluorescent nanoparticles (IFNs) were used to capture and identify the target molecules simultaneously. A good linear relationship between the fluorescence intensity and the concentration of PSA (0.1-100ng/mL) with a detection limit 27pg/mL was obtained.

  12. Highly selective and sensitive fluorescence chemosensor for the detection of palladium species based on Tsuji-Trost reaction.

    PubMed

    Xu, Zhong-Yong; Li, Jing; Guan, Su; Zhang, Lei; Dong, Chang-Zhi

    2015-09-05

    A new chemosensor 7-nitro-2,1,3-benzoxadiazole-4-allyl-N-(thiophen-2-ylmethyl)carbamate (NBDTC) was synthesized and utilized for palladium detection based on the Tsuji-Trost reaction. NBDTC displayed specific and ratiometric fluorescent responses toward palladium species. The chemosensor showed more than 50-fold enhancement in fluorescence intensity with the presence of PEG400 and palladium because NBDTC can be transformed to NBDT under palladium-catalyzing Tsuji-Trost reaction. NBDTC displayed high selectivity and sensitivity for palladium species with the detection limit of 1.13×10(-9) M.

  13. Rhodamine 6G hydrazone bearing thiophene unit: A highly sensitive and selective off-on fluorescent chemosensor for Al3+

    NASA Astrophysics Data System (ADS)

    Wu, Wei-Na; Mao, Pan-Dong; Wang, Yuan; Zhao, Xiao-Lei; Jia, Lei; Xu, Zhou-Qing

    2016-10-01

    A rhodamine derivative (R1) has been synthesized by a hydrazone formation of rhodamine 6G hydrazide with 3-methylthiophene-2-carbaldehyde, which exhibits high selectivity and sensitivity as an "off-on" fluorescent sensor toward Al3+ in water containing media. The binding process was confirmed by UV-vis absorption, fluorescence measurements, mass spectroscopy and DFT calculation. The probe functions by Al3+ induced hydrolytic cleavage of the imine-bond to produce an intense rhodamine-based emission. To test the practical use of the probe, the determination of Al3+ in real water samples was also evaluated.

  14. Fluorescent probe based on heteroatom containing styrylcyanine: pH-sensitive properties and bioimaging in vivo.

    PubMed

    Yang, Xiaodong; Gao, Ya; Huang, Zhibing; Chen, Xiaohui; Ke, Zhiyong; Zhao, Peiliang; Yan, Yichen; Liu, Ruiyuan; Qu, Jinqing

    2015-01-01

    A novel fluorescent probe based on heteroatom containing styrylcyanine is synthesized. The fluorescence of probe is bright green in basic and neutral media but dark orange in strong acidic environments, which could be reversibly switched. Such behavior enables it to work as a fluorescent pH sensor in the solution state and a chemosensor for detecting acidic and basic volatile organic compounds. Analyses by NMR spectroscopy confirm that the protonation or deprotonation of pyridinyl moiety is responsible for the sensing process. In addition, the fluorescent microscopic images of probe in live cells and zebrafish are achieved successfully, suggesting that the probe has good cell membrane permeability and low cytotoxicity.

  15. MicroProbe Small Unmanned Aerial System

    NASA Technical Reports Server (NTRS)

    Bland, Geoffrey; Miles, Ted

    2012-01-01

    The MicroProbe unmanned aerial system (UAS) concept incorporates twin electric motors mounted on the vehicle wing, thus enabling an aerodynamically and environmentally clean nose area for atmospheric sensors. A payload bay is also incorporated in the fuselage to accommodate remote sensing instruments. A key feature of this concept is lightweight construction combined with low flying speeds to minimize kinetic energy and associated hazards, as well as maximizing spatial resolution. This type of aerial platform is needed for Earth science research and environmental monitoring. There were no vehicles of this type known to exist previously.

  16. Laser Microprobe Mass Spectrometry 1: Basic Principles and Performance Characteristics.

    ERIC Educational Resources Information Center

    Denoyer, Eric; And Others

    1982-01-01

    Describes the historical development, performance characteristics (sample requirements, analysis time, ionization characteristics, speciation capabilities, and figures of merit), and applications of laser microprobe mass spectrometry. (JN)

  17. Ratiometric fluorescence and mesoporous structured imprinting nanoparticles for rapid and sensitive detection 2,4,6-trinitrophenol.

    PubMed

    Li, Ming; Liu, Haijian; Ren, Xueqin

    2017-03-15

    The present study reports the fabrication of mesoporous-structured ratiometric molecularly imprinted sensors using a combined surface-imprinted and ratiometric fluorescence method. The sensors were subsequently examined in the selective and sensitive determination of 2,4,6-trinitrophenol (TNP). In the preparation of the ratiometric system, the reference dye CdTe quantum dots were embedded in silica core particles via the Stöber method; the functional target sensitive dye AAMBT&SiO2, which was obtained via polymerization of 2-acrylamide-6-methoxybenzothiazole (AAMBT) with allyltriethoxysilane, was embedded in the mesoporous silica shell. In the surface imprinting process, cetyltrimethylammonium bromide was employed to create mesoporous-structured silica to promote quenching of AAMBT by TNP via resonance energy transfer, thereby enhancing the sensitivity of the sensor. Under optimum conditions, the ratiometric fluorescence molecularly imprinted polymer sensors achieved a detection limit of 43nM within 3min. The practical application of the developed sensor in real water samples was successfully demonstrated through analysis of TNP in water samples, achieving satisfactory recoveries of 92-104%. Thus, a convenient and practical method for preparing highly selective and sensitive ratiometric fluorescence sensors is presented herein, providing a prospective method for rapid trace pollutants analysis in complex water samples.

  18. A Highly Sensitive Fluorescent Sensor for Palladium and Direct Imaging of Its Ecotoxicity in Living Model Organisms.

    PubMed

    Liu, Fei; Du, Juan; Xu, Meiying; Sun, Guoping

    2016-01-01

    Rhodamine is an ideal platform for fluorescence probes owing to its spiro-lactam framework and excellent photochemical properties. Herein, a novel rhodamine-based palladium fluorescent chemosensor, Rd-Eb, showing a fast response time (3 min), high sensitivity for palladium species over other ions, and a low detection limit (1.91×10(-7)  m), was synthesized. It can act as an obvious colorimetric as well as a fluorescent "off/on" sensor for Pd(2+) . In addition, it is also an excellent sensor for in vivo imaging of Pd(2+) in zebra fish and Daphnia magna, illuminating the impact of palladium on organisms at different growth stages with respect to biological toxicology.

  19. Detection of acid moisture in photovoltaic modules using a dual wavelength pH-sensitive fluorescent dye

    NASA Astrophysics Data System (ADS)

    Asaka, Takashi; Iwami, Kentaro; Taguchi, Atsushi; Umeda, Norihiro; Masuda, Atsushi

    2014-01-01

    The formation of acetic acid via the penetration of moisture into ethylene vinyl acetate (EVA) in photovoltaic (PV) modules is cited as the main reason for PV modules’ degradation. Currently, there is no effective method for detecting acetic moisture in PV modules. We proposed a simple method for detecting acid moisture in PV modules using a dual-wavelength pH-sensitive dye that measures pH by the ratio of the intensities of two peaks in the fluorescence spectra of the dye. We detected the pH change caused by acetic acid with the change in the intensity ratio of the fluorescence spectra of the dried dye. Furthermore, we observed that the dry fluorescent dye is heat resistant to withstand the lamination process for the manufacturing of PV modules, and has good long-term durability.

  20. Fluorescence polarization immunoassays for rapid, accurate and sensitive determination of mycotoxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Fluorescence polarization immunoassay (FPIA) is a type of homogeneous assay. For low molecular weight antigens, such as mycotoxins, it is based on the competition between an unlabeled antigen and its fluorescent-labeled derivative (tracer) for an antigen-specific antibody. The antigen content is det...

  1. Key Structural Elements of Unsymmetrical Cyanine Dyes for Highly Sensitive Fluorescence Turn-On DNA Probes.

    PubMed

    Uno, Kakishi; Sasaki, Taeko; Sugimoto, Nagisa; Ito, Hideto; Nishihara, Taishi; Hagihara, Shinya; Higashiyama, Tetsuya; Sasaki, Narie; Sato, Yoshikatsu; Itami, Kenichiro

    2017-01-17

    Unsymmetrical cyanine dyes, such as thiazole orange, are useful for the detection of nucleic acids with fluorescence because they dramatically enhance the fluorescence upon binding to nucleic acids. Herein, we synthesized a series of unsymmetrical cyanine dyes and evaluated their fluorescence properties. A systematic structure-property relationship study has revealed that the dialkylamino group at the 2-position of quinoline in a series of unsymmetrical cyanine dyes plays a critical role in the fluorescence enhancement. Four newly designed unsymmetrical cyanine dyes showed negligible intrinsic fluorescence in the free state and strong fluorescence upon binding to double-stranded DNA (dsDNA) with a quantum yield of 0.53 to 0.90, which is 2 to 3 times higher than previous unsymmetrical cyanine dyes. A detailed analysis of the fluorescence lifetime revealed that the dialkylamino group at the 2-position of quinoline suppressed nonradiative decay in favor of increased fluorescence quantum yield. Moreover, these newly developed dyes were able to stain the nucleus specifically in fixed HeLa cells examined by using a confocal laser-scanning microscope.

  2. A nanostructured aluminum oxide-based microfluidic device for enhancing immunoassay's fluorescence and detection sensitivity.

    PubMed

    Li, Xiang; Yin, Haocheng; Que, Long

    2014-10-01

    A nanostructured aluminum oxide (NAO)-based fluorescence biosensing platform with a programmable sample delivery microfluidic interface is reported. The NAO-based fluorescence sensor can tremendously enhance the fluorescence signals, typically up to 100 × or more, over the glass substrate. The programmable sample delivery microfluidic interface, which is integrated with the NAO-based sensors, can automatically generate and deliver a series of different concentrations of the biological samples to each individual sensor. Hence it can facilitate the fluorescence-based biodetection and analysis for high throughput applications. Using Protein A and fluorophore-labeled Immunoglobulin G (IgG) as models, the binding between them on this platform have been demonstrated. It has been shown that the IgG of programmable concentrations can be delivered to individual sensor using the microfluidic interface and confirmed by the fluorescence images. Using current NAO-based fluorescence sensors without any optimization, the detectable concentration of IgG can be as low as 20 pg/mm(2) using a conventional fluorescence microscope. Due to its inexpensive fabrication process, this technology could provide a disposable technical platform for fluorescence-based sensing and analysis.

  3. Strain in UHMWPE for orthopaedic use studied by Raman microprobe spectroscopy.

    PubMed

    Kyomoto, Masayuki; Miwa, Yasutake; Pezzotti, Giuseppe

    2007-01-01

    Ultra-high-molecular-weight polyethylene (UHMWPE) has been the most popular bearing material against both metal and ceramic counter-faces in total hip and knee joint replacements. Therefore, it is desirable to clarify the complex phenomena occurring both in vivo and in vitro, using highly sensitive analytical techniques. However, conventional analytical techniques used so far suffer from destructive measurements, lack of precision and/or intricate techniques. In the present study, the physical and chemical properties of both conventional UHMWPE (PE) and highly cross-linked UHMWPE (CLPE) were investigated by Raman microprobe spectroscopy, which combines the advantages of high precision and non-destructive measurements. It was found that the strain of UHMWPE can be evaluated by a change in the full width at half maximum (FWHM) of a selected Raman band (located at around 1127 cm(-1)), and that these spectroscopic strain coefficients were (0.42 +/- 0.01) x 10(-2) cm(-1)/% elongation and (0.48 +/- 0.01) x 10(-2) cm(-1)/% elongation for PE and CLPE (100 kGy), respectively. The difference in the crystalline nature between PE and CLPE was also confirmed by Raman microprobe spectroscopy. In addition, the Raman microprobe spectroscopy technique enabled us to obtain hyperspectral images of strain and crystallinity on a microscopic scale. Thus, Raman microprobe spectroscopy is a very effective method for analyzing UHMWPE for orthopaedic use.

  4. Environment-sensitive fluorophores with benzothiadiazole and benzoselenadiazole structures as candidate components of a fluorescent polymeric thermometer.

    PubMed

    Uchiyama, Seiichi; Kimura, Kohki; Gota, Chie; Okabe, Kohki; Kawamoto, Kyoko; Inada, Noriko; Yoshihara, Toshitada; Tobita, Seiji

    2012-07-27

    An environment-sensitive fluorophore can change its maximum emission wavelength (λ(em)), fluorescence quantum yield (Φ(f)), and fluorescence lifetime in response to the surrounding environment. We have developed two new intramolecular charge-transfer-type environment-sensitive fluorophores, DBThD-IA and DBSeD-IA, in which the oxygen atom of a well-established 2,1,3-benzoxadiazole environment-sensitive fluorophore, DBD-IA, has been replaced by a sulfur and selenium atom, respectively. DBThD-IA is highly fluorescent in n-hexane (Φ(f) =0.81, λ(em) =537 nm) with excitation at 449 nm, but is almost nonfluorescent in water (Φ(f) =0.037, λ(em) =616 nm), similarly to DBD-IA (Φ(f) =0.91, λ(em) =520 nm in n-hexane; Φ(f) =0.027, λ(em) =616 nm in water). A similar variation in fluorescence properties was also observed for DBSeD-IA (Φ(f) =0.24, λ(em) =591 nm in n-hexane; Φ(f) =0.0046, λ(em) =672 nm in water). An intensive study of the solvent effects on the fluorescence properties of these fluorophores revealed that both the polarity of the environment and hydrogen bonding with solvent molecules accelerate the nonradiative relaxation of the excited fluorophores. Time-resolved optoacoustic and phosphorescence measurements clarified that both intersystem crossing and internal conversion are involved in the nonradiative relaxation processes of DBThD-IA and DBSeD-IA. In addition, DBThD-IA exhibits a 10-fold higher photostability in aqueous solution than the original fluorophore DBD-IA, which allowed us to create a new robust molecular nanogel thermometer for intracellular thermometry.

  5. A sensitive and selective determination method of histamine by HPLC with intramolecular excimer-forming derivatization and fluorescence detection.

    PubMed

    Yoshitake, Takashi; Ichinose, Fumio; Yoshida, Hideyuki; Todoroki, Ken-ichiro; Kehr, Jan; Inoue, Osamu; Nohta, Hitoshi; Yamaguchi, Masatoshi

    2003-12-01

    A highly sensitive, selective and simple method is described for the determination of histamine by high-performance liquid chromatography (HPLC) with fluorescence detection. The method is based on an intramolecular excimer-forming fluorescence derivatization of histamine with 4-(1-pyrene)butyric acid N-hydroxysuccinimide ester (PSE), followed by reversed-phase HPLC. Histamine, having two amino moieties in a molecule, was converted to the dipyrene-labeled derivative by reaction with PSE. The derivative afforded intramolecular excimer fluorescence (450-540 nm), which can clearly be discriminated from the monomer fluorescence (370-420 nm) emitted from PSE. Typically, a 10 micro L sample solution was mixed with 100 micro L of derivatization reagent solution, which was a mixture of 0.5 mm PSE in acetonitrile and 0.5 mm potassium carbonate in water (8:2, v/v). The derivatization was carried out at 100 degrees C for 90 min. The PSE derivative of histamine could be separated by reversed-phase ODS column with isocratic elution using acetonitrile:water (82:18, v/v) containing 0.03% triethylamine. The detection limit (singnal-to-noise ratio = 3) of histamine was 0.5 fmol for a 30 micro L injection. The method was successfully applied to the determination of histamine in human urine, and had enough selectivity and sensitivity for urinary histamine quantification.

  6. Peptide-Induced AIEgen Self-Assembly: A New Strategy to Realize Highly Sensitive Fluorescent Light-Up Probes.

    PubMed

    Han, Aitian; Wang, Huaimin; Kwok, Ryan T K; Ji, Shenglu; Li, Jun; Kong, Deling; Tang, Ben Zhong; Liu, Bin; Yang, Zhimou; Ding, Dan

    2016-04-05

    Fluorescent light-up probes with aggregation-induced emission (AIE) characteristics have recently attracted great research interest due to their intelligent fluorescence activation mechanism and excellent photobleaching resistance. In this work, we report a new, simple, and generic strategy to design and prepare highly sensitive AIE fluorescent light-up bioprobe through facile incorporation of a self-assembling peptide sequence GFFY between the recognition element and the AIE luminogen (AIEgen). After the bioprobes respond to the targets, the peptide GFFY is capable of inducing the ordered self-assembly of AIEgens, yielding close and tight intermolecular steric interactions to restrict the intramolecular motions of AIEgens for excellent signal output. Using two proof-of-concepts, we have demonstrated that self-assembling peptide-incorporating AIE light-up probes show much higher sensitivity in sensing the corresponding targets in both solutions and cancer cells as compared to those without GFFY induced self-assembly. Taking the probe TPE-GFFYK(DVEDEE-Ac), for example, a detection limit as low as 0.54 pM can be achieved for TPE-GFFYK(DVEDEE-Ac) in caspase-3 detection, which is much lower than that of TPE-K(DVED-Ac) (3.50 pM). This study may inspire new insights into the design of advanced fluorescent molecular probes.

  7. Supramolecular spectrally encoded microgels with double strand probes for absolute and direct miRNA fluorescence detection at high sensitivity.

    PubMed

    Causa, Filippo; Aliberti, Anna; Cusano, Angela M; Battista, Edmondo; Netti, Paolo A

    2015-02-11

    We present novel microgels as a particle-based suspension array for direct and absolute microRNA (miRNA) detection. The microgels feature a flexible molecular architecture, antifouling properties, and enhanced sensitivity with a large dynamic range of detection. Specifically, they possess a core-shell molecular architecture with two different fluorescent dyes for multiplex spectral analyses and are endowed with a fluorescent probe for miRNA detection. Encoding and detection fluorescence signals are distinguishable by nonoverlapping emission spectra. Tunable fluorescence probe conjugation and emission confinement on single microgels allow for ultrasensitive miRNA detection. Indeed, the suspension array has high selectivity and sensitivity with absolute quantification, a detection limit of 10(-15) M, a dynamic range from 10(-9) to 10(-15) M, and higher accuracy than qRT-PCR. The antifouling properties of the microgels also permit the direct measurement of miRNAs in serum, without sample pretreatment or target amplification. A multiplexed assay has been tested for a set of miRNAs chosen as cancer biomarkers.

  8. l-Tryptophan-capped carbon quantum dots for the sensitive and selective fluorescence detection of mercury ion in aqueous solution

    NASA Astrophysics Data System (ADS)

    Wan, Xuejuan; Li, Shifeng; Zhuang, Lulu; Tang, Jiaoning

    2016-07-01

    l-Tryptophan-capped carbon quantum dots ( l-CQDs) were facilely synthesized through "green" methodology, and the obtained material was utilized as a sensitive and selective fluorescence sensor for mercury ion (Hg2+) in pure aqueous solutions. Carboxyl-functionalized CQDs were first green synthesized by a one-step hydrothermal route, and l-tryptophan was then attached to CQDs via direct surface condensation reaction in aqueous solution at room temperature. The as-synthesized l-CQDs had an average size of ca. 5 nm with a good dispersity in water, and exhibited a favorable selectivity for Hg2+ ions over a range of other common metal cations in aqueous solution (10 mM PBS buffer, pH 6.0). Upon the addition of Hg2+, a complete fluorescence quenching (ON-OFF switching) of l-CQDs was evident from the fluorescence titration experiment, and the fluorescence detection limit of Hg2+ was calculated to be 11 nM, which indicated that the obtained environmentally friendly l-CQDs had sensitive detection capacity for Hg2+ in aqueous solution.

  9. Molecular switching fluorescence based high sensitive detection of label-free C-reactive protein on biochip.

    PubMed

    Islam, Md Shahinul; Yu, Hyunung; Lee, Hee Gu; Kang, Seong Ho

    2010-11-15

    A novel detection technique on biochip for the quantification of label-free C-reactive protein (CRP) based on molecular switching of fluorescence (MSF) is demonstrated by total internal reflection fluorescence microscopy. It alters fluorescence intensity of fluoreseinamine isomer 1 (FAI) upon binding with its specific ligand, O-phosphorylethanolamine (PEA). In the MSF-based detection, FAI was used as an ink, printed on a 3-glycidoxypropyl-trimethoxysilane (GPTS)-coated glass coverslip. With the addition of GPTS conjugated PEA solution to the FAI-printed coverslip, the fluorescence intensity was remarkably decreased. Addition of CRP increased fluorescence intensity linearly in the range of 800 aM to 500 fM (R=0.997). The MSF-based biochip assay for the estimation of CRP in human sera showed ∼200 times increased detection sensitivity in less than a third of the time to obtain results using a conventional enzyme-linked immunosorbent assay. This biochip detection is a promising new technique for the quantification of CRP molecules from trace amounts of clinical samples.

  10. Label-free fluorescence strategy for sensitive detection of exonuclease activity using SYBR Green I as probe.

    PubMed

    Xu, Min; Li, Baoxin

    2015-01-01

    A label-free and sensitive fluorescence assay for exonuclease activity is developed using commercially available SYBR Green I (SG) dye as signal probe. A proof-of-concept of this assay has been demonstrated by using exonuclease III (Exo III) as a model enzyme. In this assay, double-stranded DNA (dsDNA) can bind SG, resulting in a strong fluorescence signal of SG. Upon the addition of Exo III, dsDNA would be digested, and SG emits very weak fluorescence. Thus, Exo III activity can be facilely measured with a simple fluorescence reader. This method has a linear detection range from 1 U/mL to 200 U/mL with a detection limit of 0.7 U/mL. This label-free approach is selective, simple, convenient and cost-efficient without any complex DNA sequence design or fluorescence dye label. The method not only provides a platform for monitoring activity and inhibition of exonuclease but also shows great potential in biological process researches, drug discovery, and clinic diagnostics.

  11. Environment-Sensitive Fluorescent Probe for the Human Ether-a-go-go-Related Gene Potassium Channel

    PubMed Central

    2016-01-01

    A novel environment-sensitive probe S2 with turn-on switch for Human Ether-a-go-go-Related Gene (hERG) potassium channel was developed herein. After careful evaluation, this fluorescent probe showed high binding affinity with hERG potassium channel with an IC50 value of 41.65 nM and can be well applied to hERG channel imaging or cellular distribution study for hERG channel blockers. Compared with other imaging techniques, such as immunofluorescence and fluorescent protein-based approaches, this method is convenient and affordable, especially since a washing procedure is not needed. Meanwhile, this environment-sensitive turn-on design strategy may provide a good example for the probe development for these targets that have no reactive or catalytic activity. PMID:26730746

  12. Environment-Sensitive Fluorescent Probe for the Human Ether-a-go-go-Related Gene Potassium Channel.

    PubMed

    Liu, Zhenzhen; Jiang, Tianyu; Wang, Beilei; Ke, Bowen; Zhou, Yubin; Du, Lupei; Li, Minyong

    2016-02-02

    A novel environment-sensitive probe S2 with turn-on switch for Human Ether-a-go-go-Related Gene (hERG) potassium channel was developed herein. After careful evaluation, this fluorescent probe showed high binding affinity with hERG potassium channel with an IC50 value of 41.65 nM and can be well applied to hERG channel imaging or cellular distribution study for hERG channel blockers. Compared with other imaging techniques, such as immunofluorescence and fluorescent protein-based approaches, this method is convenient and affordable, especially since a washing procedure is not needed. Meanwhile, this environment-sensitive turn-on design strategy may provide a good example for the probe development for these targets that have no reactive or catalytic activity.

  13. A novel fluorescent turn-on biosensor based on QDs@GSH-GO fluorescence resonance energy transfer for sensitive glutathione S-transferase sensing and cellular imaging.

    PubMed

    Chang, Lifang; He, Xiwen; Chen, Langxing; Zhang, Yukui

    2017-03-17

    A novel fluorescent turn-on biosensor based on fluorescence resonance energy transfer (FRET) from GSH functionalized Mn-doped ZnS QDs to graphene oxide (GO) was constructed to determine glutathione S-transferases (GSTs) in live cells and human urine. The QDs@GSH is adsorbed on the GO surface via hydrogen bonding interaction between the GSH on the surface of QDs@GSH and GO, and as a result, fluorescence quenching of the QDs@GSH takes place because of FRET. The FRET efficiency from QDs@GSH to GO was calculated to be 86.3%. However, in the presence of GSTs, the FRET process could be inhibited by the specific interaction between the GSH on the surface of QDs@GSH and GSTs, which would keep the QDs@GSH far away from the GO surface, leading to the recovery of the fluorescence. The proposed sensor exhibited high sensitivity, selectivity, and excellent specificity in the buffer, live cells and human urine for the detection of GSTs. Under the physiological conditions (pH 7.4), dissociation constants and the detection limit of GST and ATP6 V1F (a GST-tagged protein) were estimated to be 8.0 × 10(-9) M, 2.1 × 10(-10) M and 3.5 × 10(-9) M, 7.2 × 10(-11) M, respectively. The presented method has been successfully utilized for the determination of the GSTs in live cells and human urine without any complicated pretreatment and the recovery was in the range of 80%-90%.

  14. Analysis of biological materials using a nuclear microprobe

    NASA Astrophysics Data System (ADS)

    Mulware, Stephen Juma

    The use of nuclear microprobe techniques including: Particle induced x-ray emission (PIXE) and Rutherford backscattering spectrometry (RBS) for elemental analysis and quantitative elemental imaging of biological samples is especially useful in biological and biomedical research because of its high sensitivity for physiologically important trace elements or toxic heavy metals. The nuclear microprobe of the Ion Beam Modification and Analysis Laboratory (IBMAL) has been used to study the enhancement in metal uptake of two different plants. The roots of corn (Zea mays) have been analyzed to study the enhancement of iron uptake by adding Fe (II) or Fe(III) of different concentrations to the germinating medium of the seeds. The Fe uptake enhancement effect produced by lacing the germinating medium with carbon nanotubes has also been investigated. The aim of this investigation is to ensure not only high crop yield but also Fe-rich food products especially from calcareous soil which covers 30% of world's agricultural land. The result will help reduce iron deficiency anemia, which has been identified as the leading nutritional disorder especially in developing countries by the World Health Organization. For the second plant, Mexican marigold (Tagetes erecta ), the effect of an arbuscular mycorrhizal fungi (Glomus intraradices ) for the improvement of lead phytoremediation of lead contaminated soil has been investigated. Phytoremediation provides an environmentally safe technique of removing toxic heavy metals (like lead), which can find their way into human food, from lands contaminated by human activities like mining or by natural disasters like earthquakes. The roots of Mexican marigold have been analyzed to study the role of arbuscular mycorrhizal fungi in enhancement of lead uptake from the contaminated rhizosphere.

  15. Influence of complexing ion on the fluorescence sensitization efficiency for oxazine dyes in nanoparticles of Sc, Eu, Sm, and Lu diketonates

    NASA Astrophysics Data System (ADS)

    Mironov, L. Yu.

    2016-12-01

    The fluorescence sensitization regularities have been investigated for oxazine dyes (Nile blue, cresyl violet (oxazine-9), and oxazine-170) in nanoparticles of complexes of 2-naphthoyltrifluoroacetone with trivalent ions of rare-earth metals. The fluorescence sensitization efficiencies of dyes in nanoparticles from the Sc(III), Eu(III), Sm(III), and Lu(III) complexes are compared. It is shown that the fluorescence sensitization efficiencies of dyes in nanoparticles from the Sc(III), Eu(III), and Sm(III) complexes has similar values and greatly exceed that for nanoparticles from the Lu(III) complexes. The quantum yields of sensitized fluorescence are determined for dyes in nanoparticles from the Sc(III), Eu(III), and Sm(III) complexes. The nanoparticles doped with oxazine-170 from Eu(III) complexes are found to have the strongest fluorescence.

  16. A practical and highly sensitive C3N4-TYR fluorescent probe for convenient detection of dopamine

    NASA Astrophysics Data System (ADS)

    Li, Hao; Yang, Manman; Liu, Juan; Zhang, Yalin; Yang, Yanmei; Huang, Hui; Liu, Yang; Kang, Zhenhui

    2015-07-01

    The C3N4-tyrosinase (TYR) hybrid is a highly accurate, sensitive and simple fluorescent probe for the detection of dopamine (DOPA). Under optimized conditions, the relative fluorescence intensity of C3N4-TYR is proportional to the DOPA concentration in the range from 1 × 10-3 to 3 × 10-8 mol L-1 with a correlation coefficient of 0.995. In the present system, the detection limit achieved is as low as 3 × 10-8 mol L-1. Notably, these quantitative detection results for clinical samples are comparable to those of high performance liquid chromatography. Moreover, the enzyme-encapsulated C3N4 sensing arrays on both glass slide and test paper were evaluated, which revealed sensitive detection and excellent stability. The results reported here provide a new approach for the design of a multifunctional nanosensor for the detection of bio-molecules.The C3N4-tyrosinase (TYR) hybrid is a highly accurate, sensitive and simple fluorescent probe for the detection of dopamine (DOPA). Under optimized conditions, the relative fluorescence intensity of C3N4-TYR is proportional to the DOPA concentration in the range from 1 × 10-3 to 3 × 10-8 mol L-1 with a correlation coefficient of 0.995. In the present system, the detection limit achieved is as low as 3 × 10-8 mol L-1. Notably, these quantitative detection results for clinical samples are comparable to those of high performance liquid chromatography. Moreover, the enzyme-encapsulated C3N4 sensing arrays on both glass slide and test paper were evaluated, which revealed sensitive detection and excellent stability. The results reported here provide a new approach for the design of a multifunctional nanosensor for the detection of bio-molecules. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr03316k

  17. Fluorescent probes sensitive to changes in the cholesterol-to-phospholipids molar ratio in human platelet membranes during atherosclerosis

    NASA Astrophysics Data System (ADS)

    Posokhov, Yevgen

    2016-09-01

    Environment-sensitive fluorescent probes were used for the spectroscopic visualization of pathological changes in human platelet membranes during cerebral atherosclerosis. It has been estimated that the ratiometric probes 2-(2‧-hydroxyphenyl)-5-phenyl-1,3,4-oxadiazole and 2-phenyl-phenanthr[9,10]oxazole can detect changes in the cholesterol-to-phospholipids molar ratio in human platelet membranes during the disease.

  18. High sensitivity fluorescent single particle and single molecule detection apparatus and method

    DOEpatents

    Mathies, Richard A.; Peck, Konan; Stryer, Lubert

    1990-01-01

    Apparatus is described for ultrasensitive detection of single fluorescent particles down to the single fluorescent molecule limit in a fluid or on a substrate comprising means for illuminating a predetermined volume of the fluid or area of the substrate whereby to emit light including background light from the fluid and burst of photons from particles residing in the area. The photon burst is detected in real time to generate output representative signal. The signal is received and the burst of energy from the fluorescent particles is distinguished from the background energy to provide an indication of the number, location or concentration of the particles or molecules.

  19. Environment-sensitive fluorescent probe: a benzophosphole oxide with an electron-donating substituent.

    PubMed

    Yamaguchi, Eriko; Wang, Chenguang; Fukazawa, Aiko; Taki, Masayasu; Sato, Yoshikatsu; Sasaki, Taeko; Ueda, Minako; Sasaki, Narie; Higashiyama, Tetsuya; Yamaguchi, Shigehiro

    2015-04-07

    Electron-donating aryl groups were attached to electron-accepting benzophosphole skeletons. Among several derivatives thus prepared, one benzophosphole oxide was particularly interesting, as it retained high fluorescence quantum yields even in polar and protic solvents. This phosphole-based compound exhibited a drastic color change of its fluorescence spectrum as a function of the solvent polarity, while the absorption spectra remained virtually unchanged. Capitalizing on these features, this phosphole-based compound was used to stain adipocytes, in which the polarity of subcellular compartments could then be discriminated on the basis of the color change of the fluorescence emission.

  20. Enhanced sensitivity of pHluorin-based monitoring of intracellular pH changes achieved through synchronously scanned fluorescence spectra.

    PubMed

    Plášek, Jaromír; Melcrová, Adéla; Gášková, Dana

    2015-10-06

    Since its introduction in 1998, genetically encoded pH-sensitive sensor ratiometric pHluorin proved to be a valuable tool for cell physiology studies. Here, we show how the sensitivity of pHluorin-based monitoring of intracellular pH changes performed with cell suspensions can be enhanced by using synchronously scanned fluorescence spectroscopy. In the suspensions of S. cerevisiae cells subjected to varying extracellular pH values, we have been able to measure statistically significant changes in intracellular pH of less than 0.1 unit, which were not detectable using a standard ratiometric approach.

  1. A sensitive quantum dots-based "OFF-ON" fluorescent sensor for ruthenium anticancer drugs and ctDNA.

    PubMed

    Huang, Shan; Zhu, Fawei; Qiu, Hangna; Xiao, Qi; Zhou, Quan; Su, Wei; Hu, Baoqing

    2014-05-01

    In this contribution, a simple and sensitive fluorescent sensor for the determination of both the three ruthenium anticancer drugs (1 to 3) and calf thymus DNA (ctDNA) was established based on the CdTe quantum dots (QDs) fluorescence "OFF-ON" mode. Under the experimental conditions, the fluorescence of CdTe QDs can be effectively quenched by ruthenium anticancer drugs because of the surface binding of these drugs on CdTe QDs and the subsequent photoinduced electron transfer (PET) process from CdTe QDs to ruthenium anticancer drugs, which render the system into fluorescence "OFF" status. The system can then be "ON" after the addition of ctDNA which brought the restoration of CdTe QDs fluorescence intensity, since ruthenium anticancer drugs broke away from the surface of CdTe QDs and inserted into double helix structure of ctDNA. The fluorescence quenching effect of the CdTe QDs-ruthenium anticancer drugs systems was mainly concentration dependent, which could be used to detect three ruthenium anticancer drugs. The limits of detection were 5.5 × 10(-8) M for ruthenium anticancer drug 1, 7.0 × 10(-8) M for ruthenium anticancer drug 2, and 7.9× 10(-8) M for ruthenium anticancer drug 3, respectively. The relative restored fluorescence intensity was directly proportional to the concentration of ctDNA in the range of 1.0 × 10(-8) M ∼ 3.0 × 10(-7) M, with a correlation coefficient (R) of 0.9983 and a limit of detection of 1.1 × 10(-9) M. The relative standard deviation (RSD) for 1.5 × 10(-7) M ctDNA was 1.5% (n = 5). There was almost no interference to some common chemical compounds, nucleotides, amino acids, and proteins. The proposed method was applied to the determination of ctDNA in three synthetic samples with satisfactory results. The possible reaction mechanism of CdTe QDs fluorescence "OFF-ON" was further investigated. This simple and sensitive approach possessed some potential applications in the investigation of interaction between drug molecules and DNA.

  2. Sensitive and selective detection of Hg2+ and Cu2+ ions by fluorescent Ag nanoclusters synthesized via a hydrothermal method

    NASA Astrophysics Data System (ADS)

    Liu, Jing; Ren, Xiangling; Meng, Xianwei; Fang, Zheng; Tang, Fangqiong

    2013-09-01

    An easily prepared fluorescent Ag nanoclusters (Ag NCs) probe for the sensitive and selective detection of Hg2+ and Cu2+ ions was developed here. The Ag NCs were synthesized by using polymethacrylic acid sodium salt as a template via a convenient hydrothermal process. The as-prepared fluorescent Ag NCs were monodispersed, uniform and less than 2 nm in diameter, and can be quenched in the presence of mercury (Hg2+) or copper (Cu2+) ions. Excellent linear relationships existed between the quenching degree of the Ag NCs and the concentrations of Hg2+ or Cu2+ ions in the range of 10 nM to 20 μM or 10 nM to 30 μM, respectively. By using ethylenediaminetetraacetate (EDTA) as the masking agent of Cu2+, Hg2+ was exclusively detected in coexistence with Cu2+ with high sensitivity (LOD = 10 nM), which also provided a reusable detection method for Cu2+. Furthermore, the different quenching phenomena caused by the two metals ions such as changes in visible colour, shifts of UV absorbance peaks and changes in size of Ag NCs make it easy to distinguish between them. Therefore the easily synthesized fluorescent Ag NCs may have great potential as Hg2+ and Cu2+ ions sensors.An easily prepared fluorescent Ag nanoclusters (Ag NCs) probe for the sensitive and selective detection of Hg2+ and Cu2+ ions was developed here. The Ag NCs were synthesized by using polymethacrylic acid sodium salt as a template via a convenient hydrothermal process. The as-prepared fluorescent Ag NCs were monodispersed, uniform and less than 2 nm in diameter, and can be quenched in the presence of mercury (Hg2+) or copper (Cu2+) ions. Excellent linear relationships existed between the quenching degree of the Ag NCs and the concentrations of Hg2+ or Cu2+ ions in the range of 10 nM to 20 μM or 10 nM to 30 μM, respectively. By using ethylenediaminetetraacetate (EDTA) as the masking agent of Cu2+, Hg2+ was exclusively detected in coexistence with Cu2+ with high sensitivity (LOD = 10 nM), which also provided a

  3. A sensitive method based on fluorescence-detected circular dichroism for protein local structure analysis.

    PubMed

    Nehira, Tatsuo; Ishihara, Kaoru; Matsuo, Koichi; Izumi, Shunsuke; Yamazaki, Takeshi; Ishida, Atsuhiko

    2012-11-15

    We report an improved fluorescence-detected circular dichroism (FDCD)-based analytical method that is useful for probing protein three-dimensional structures. The method uses a novel FDCD device with an ellipsoidal mirror that functions on a standard circular dichroism (CD) spectrometer and eliminates all artifacts. Our experiments demonstrated three important findings. First, the method is applicable to any proteins either by using intrinsic fluorescence derived from tryptophan residues or by introducing a fluorescent label onto nonfluorescent proteins. Second, by using intrinsic fluorescence, FDCD spectroscopy can detect a structural change in the tertiary structure of metmyoglobin due to stepwise denaturation on a change in pH. Such changes could not be detected by conventional CD spectroscopy. Third, based on the typical advantages of fluorescence-based analyses, FDCD measurements enable observation of only the target proteins in a solution even in the presence of other peptides. Using our ellipsoidal mirror FDCD device, we could observe structural changes of fluorescently labeled calmodulin on binding with Ca(2+) and/or interacting with binding peptides. Because FDCD appears to reflect the protein's local structure around the fluorophore, it may provide a useful means for "pinpoint analysis" of protein structures.

  4. Chloride-sensitive MEQ fluorescence in chick embryo motoneurons following manipulations of chloride and during spontaneous network activity.

    PubMed

    Chub, Nikolai; Mentis, George Z; O'donovan, Michael J

    2006-01-01

    Intracellular Cl(-) ([Cl(-)](in)) homeostasis is thought to be an important regulator of spontaneous activity in the spinal cord of the chick embryo. We investigated this idea by visualizing the variations of [Cl(-)](in) in motoneurons retrogradely labeled with the Cl-sensitive dye 6-methoxy-N-ethylquinolinium iodide (MEQ) applied to cut muscle nerves in the isolated E10-E12 spinal cord. This labeling procedure obviated the need for synthesizing the reduced, cell-permeable dihydro-MEQ (DiH-MEQ). The specificity of motoneuron labeling was confirmed using retrograde co-labeling with Texas Red Dextran and immunocytochemistry for choline acetyltransferase (ChAT). In MEQ-labeled motoneurons, the GABA(A) receptor agonist isoguvacine (100 muM) increased somatic and dendritic fluorescence by 7.4 and 16.7%, respectively. The time course of this fluorescence change mirrored that of the depolarization recorded from the axons of the labeled motoneurons. Blockade of the inward Na(+)/K(-)/2Cl(-) co-transporter (NKCC1) with bumetanide (20 microM) or with a low-Na(+) bath solution (12 mM), increased MEQ fluorescence by 5.3 and 11.4%, respectively, consistent with a decrease of [Cl(-)](in). After spontaneous episodes of activity, MEQ fluorescence increased and then declined to the pre-episode level during the interepisode interval. The largest fluorescence changes occurred over motoneuron dendrites (19.7%) with significantly smaller changes (5.2%) over somata. Collectively, these results show that retrogradely loaded MEQ can be used to detect [Cl(-)](in) in motoneurons, that the bumetanide-sensitive NKCC1 co-transporter is at least partially responsible for the elevated [Cl(-)](in) of developing motoneurons, and that dendritic [Cl(-)](in) decreases during spontaneous episodes and recovers during the inter-episode interval, presumably due to the action of NKCC1.

  5. A convenient and label-free fluorescence "turn off-on" nanosensor with high sensitivity and selectivity for acid phosphatase.

    PubMed

    Liu, Ziping; Lin, Zihan; Liu, Linlin; Su, Xingguang

    2015-05-30

    In this study, we reported a convenient label-free fluorescence nanosensor for rapid detection of acid phosphatase on the basis of aggregation-caused quenching (ACQ) and enzymolysis approach. The selectivity nanosensor was based on the fluorescence "turn off-on" mode, which possessed high sensitivity features. The original strong fluorescence intensity of CuInS2 QDs was quenched by sodium hexametaphosphate (NaPO3)6. The high efficiency of the quenching was caused by the non-covalent binding of positively charged CuInS2 QDs to the negatively charged (NaPO3)6 through electrostatic interactions, aggregating to form a CuInS2 QDs/(NaPO3)6 complex. Adding acid phosphatase caused intense fluorescence of CuInS2 QDs/(NaPO3)6 to be recovered, and this was because of enzymolysis. (NaPO3)6 was hydrolyzed into small fragments and the high negative charge density decreased, which would weaken the strong electrostatic interactions. As a result, the quenched fluorescence "turned on". Under the optimum conditions, there was a good linear relationship between I/I0 (I and I0 were the fluorescence intensity of CuInS2 QDs/(NaPO3)6 system in the presence and absence of acid phosphatase, respectively) and acid phosphatase concentration in the range of 75-1500 nU mL(-1) with the detection limit of 9.02 nU mL(-1). The proposed nanosensor had been utilized to detect and accurately quantify acid phosphatase in human serum samples with satisfactory results.

  6. Direct, live imaging of cortical spreading depression and anoxic depolarisation using a fluorescent, voltage-sensitive dye

    PubMed Central

    Farkas, Eszter; Pratt, Rosalind; Sengpiel, Frank; Obrenovitch, Tihomir P

    2008-01-01

    Perilesion depolarisations, whether transient anoxic depolarisation (AD) or spreading depression (SD), occur in stroke models and in patients with acute brain ischaemia, but their contribution to lesion progression remains unclear. As these phenomena correspond to waves of cellular depolarisation, we have developed a technique for their live imaging with a fluorescent voltage-sensitive (VS) dye (RH-1838). Method development and validation were performed in two different preparations: chicken retina, to avoid any vascular interference; and cranial window exposing the cortical surface of anaesthetised rats. Spreading depression was produced by high-K medium, and AD by complete terminal ischaemia in rats. After dye loading, the preparation was illuminated at its excitation wavelength and fluorescence changes were recorded sequentially with a charge-coupled device camera. No light was recorded when the VS dye was omitted, ruling out the contribution of any endogenous fluorophore. With both preparations, the changes in VS dye fluorescence with SD were analogous to those of the DC (direct current) potential recorded with glass electrodes. Although some blood quenching of the emitted light was identified, the VS dye signatures of SD had a good signal-to-noise ratio and were reproducible. The changes in VS dye fluorescence associated with AD were more complex because of additional interferents, especially transient brain swelling with subsequent shrinkage. However, the kinetics of the AD-associated changes in VS dye fluorescence was also analogous to that of the DC potential. In conclusion, this method provides the imaging equivalent of electrical extracellular DC potential recording, with the SD and AD negative shifts translating directly to fluorescence increase. PMID:17971792

  7. Electrospun fibrous mats with conjugated tetraphenylethylene and mannose for sensitive turn-on fluorescent sensing of Escherichia coli.

    PubMed

    Zhao, Long; Chen, Yufei; Yuan, Jiang; Chen, Maohua; Zhang, Hong; Li, Xiaohong

    2015-03-11

    A rapid and sensitive detection of microbes in water and biological fluids is a key requirement in water and food safety, environmental monitoring, and clinical diagnosis and treatment. In the current study, electrospun polystyrene-co-maleic anhydride (PSMA) fibers with conjugated mannose and tetraphenylethylene (TPE) were developed for Escherichia coli (E. coli) detection, taking advantage of the high grafting capabilities of ultrafine fibers and the highly porous structure of the fibrous mat to entrap bacterial cells. The specific binding between mannose grafts on PSMA fibers and FimH proteins from the fimbriae of E. coli led to an efficient "turn-on" profile of TPE due to the aggregation-induced emission (AIE) effect. Poly(ethylene glycol) diamine was used as hydrophilic tethers to increase the conformational mobility of mannose grafts, indicating a more sensitive change in the fluorescence intensity against bacteria concentrations, a lower fluorescence background of fibers without bacteria incubation, and a sufficient space for bacteria binding, compared with the use of hexamethylenediamine or poly(ethylene imine) as spacers for mannose grafting. The addition of bovine serum albumin, glucose, or both of them into bacteria suspensions showed no significant changes in the fluorescence intensity of fibrous mats, indicating the anti-interference capability against these proteins and saccharides. An equation was drafted of the fluorescence intensities of fibrous mats against E. coli concentrations ranging from 10(2) to 10(5) CFU/mL. The test strip format was established on mannose-conjugated PSMA fibers after exposure to E. coli of different concentrations, providing a potential tool with a visual sensitivity of bacteria concentrations as low as 10(2) CFU/mL in a matter of minutes. This strategy may offer a capacity to be expanded to exploit electrospun fibrous mats and other carbohydrate-cell interactions for bioanalysis and biosensing of pathogenic bacteria.

  8. Aerodynamics of the Mars Microprobe Entry Vehicles

    NASA Technical Reports Server (NTRS)

    Mitcheltree, R. A.; Moss, J. N.; Cheatwood, F. M.; Greene, F. A.; Braun, R. D.

    1997-01-01

    The selection of the unique aeroshell shape for the Mars Microprobes is discussed. A description of its aerodynamics in hypersonic rarefied, hypersonic continuum, supersonic and transonic flow regimes is then presented. This description is based on Direct Simulation Monte Carlo analyses in the rarefied-flow regime, thermochemical nonequilibrium Computational Fluid Dynamics in the hypersonic regime, existing wind tunnel data in the supersonic and transonic regime, additional computational work in the transonic regime, and finally, ballistic range data. The aeroshell is shown to possess the correct combination of aerodynamic stability and drag to convert the probe's initial tumbling attitude and high velocity at atmospheric-interface into the desired surface-impact orientation and velocity.

  9. Ion microprobe, electron microprobe and cathodoluminescence data for Allende inclusions with emphasis on plagioclase chemistry

    NASA Technical Reports Server (NTRS)

    Hutcheon, I. D.; Steele, I. M.; Smith, J. V.; Clayton, R. N.

    1978-01-01

    Three Type B inclusions from the Allende meteorite have been analyzed. A grain-to-grain characterization of mineral chemistry and isotopic content was made possible by the use of a range of techniques, including luminescence and scanning electron microscopy and electron and ion microprobe analysis. Cathodoluminescence was used in fine-grained, optically opaque regions to distinguish between sub-micrometer phases, such as garnet and Si-rich material, subsequently identified by electron probe and scanning electron microscope analyses. Four types of luminescence patterns, due to twinning, primary sector zoning, alteration of boundaries and fractures, and shock effects, were identified in Allende plagioclase. Luminescence color exhibited a strong correlation with Mg content and provided a guide for an electron probe quantitative map of Mg and Na distributions. Ion microprobe studies of individual grains revealed large excesses of Mg-26.

  10. Fluorescent metal-organic framework MIL-53(Al) for highly selective and sensitive detection of Fe3+ in aqueous solution.

    PubMed

    Yang, Cheng-Xiong; Ren, Hu-Bo; Yan, Xiu-Ping

    2013-08-06

    Fluorescent metal-organic frameworks (MOFs) have received great attention in sensing application. Here, we report the exploration of fluorescent MIL-53(Al) for highly selective and sensitive detection of Fe(3+) in aqueous solution. The cation exchange between Fe(3+) and the framework metal ion Al(3+) in MIL-53(Al) led to the quenching of the fluorescence of MIL-53(Al) due to the transformation of strong-fluorescent MIL-53(Al) to weak-fluorescent MIL-53(Fe), allowing highly selective and sensitive detection of Fe(3+) in aqueous solution with a linear range of 3-200 μM and a detection limit of 0.9 μM. No interferences from 0.8 M Na(+); 0.35 M K(+); 11 mM Cu(2+); 10 mM Ni(2+); 6 mM Ca(2+), Pb(2+), and Al(3+); 5.5 mM Mn(2+); 5 mM Co(2+) and Cr(3+); 4 mM Hg(2+), Cd(2+), Zn(2+), and Mg(2+); 3 mM Fe(2+); 0.8 M Cl(-); 60 mM NO2(-) and NO3(-); 10 mM HPO4(2-), H2PO4(-), SO3(2-), SO4(2-), and HCOO(-); 8 mM CO3(2-), HCO3(-), and C2O4(2-); and 5 mM CH3COO(-) were found for the detection of 150 μM Fe(3+). The possible mechanism for the quenching effect of Fe(3+) on the fluorescence of MIL-53(Al) was elucidated by inductively coupled plasma-mass spectrometry, X-ray diffraction spectrometry, and Fourier transform infrared spectrometry. The specific cation exchange behavior between Fe(3+) and the framework Al(3+) along with the excellent stability of MIL-53(Al) allows highly selective and sensitive detection of Fe(3+) in aqueous solution. The developed method was applied to the determination of Fe(3+) in human urine samples with the quantitative spike recoveries from 98.2% to 106.2%.

  11. Imaging of sialidase activity in rat brain sections by a highly sensitive fluorescent histochemical method.

    PubMed

    Minami, Akira; Shimizu, Hirotaka; Meguro, Yuko; Shibata, Naoki; Kanazawa, Hiroaki; Ikeda, Kiyoshi; Suzuki, Takashi

    2011-09-01

    Sialidase (EC 3.2.1.18) removes sialic acid from sialoglycoconjugates. Since sialidase extracellularly applied to the rat hippocampus influences many neural functions, including synaptic plasticity and innervations of glutamatergic neurons, endogenous sialidase activities on the extracellular membrane surface could also affect neural functions. However, the distribution of sialidase activity in the brain remains unknown. To visualize extracellular sialidase activity on the membrane surface in the rat brain, acute brain slices were incubated with 5-bromo-4-chloroindol-3-yl-α-d-N-acetylneuraminic acid (X-Neu5Ac) and Fast Red Violet LB (FRV LB) at pH 7.3. After 1h, myelin-abundant regions showed intense fluorescence in the rat brain. Although the hippocampus showed weak fluorescence in the brain, mossy fiber terminals in the hippocampus showed relatively intense fluorescence. These fluorescence intensities were attenuated with a sialidase-specific inhibitor, 2,3-dehydro-2-deoxy-N-acetylneuraminic acid (DANA, 1mM). Additionally, the fluorescence intensities caused by X-Neu5Ac and FRV LB were correlated with the sialidase activity measured with 4-methylumbelliferyl-α-d-N-acetylneuraminic acid (4MU-Neu5Ac), a classical substrate for quantitative measurement of sialidase activity, in each brain region. Therefore, staining with X-Neu5Ac and FRV LB is specific for sialidase and useful for quantitative analysis of sialidase activities. The results suggest that white matter of the rat brain has intense sialidase activity.

  12. Nanocrystal clusters in combination with spectral imaging to improve sensitivity in antibody labeling applications of fluorescent nanocrystals

    NASA Astrophysics Data System (ADS)

    Maier, John S.; Panza, Janice L.; Bootman, Matt

    2007-02-01

    Composition-tunable nanocrystals are fluorescent nanoparticles with a uniform particle size and with adjustable optical characteristics. When used for optical labeling of biomolecular targets these and other nanotechnology solutions have enabled new approaches which are possible because of the high optical output, narrow spectral signal, consistent quantum efficiency across a broad emission range and long lived fluorescent behavior of the nanocrystals. When coupled with spectral imaging the full potential of multiplexing multiple probes in a complex matrix can be realized. Spectral imaging can be used to improve sensitivity of narrowband fluorophores through application of chemometric image processing techniques used to reduce the influence of autofluorescence background. Composition-tunable nanocrystals can be complexed together to form nanoclusters which have the advantage of significantly stronger signal and therefore a higher sensitivity. These nanoclusters can be targeted in biomolecular systems using standard live-cell labeling and immunohistochemistry based techniques. Composition-tunable nanocrystals and nanoclusters have comparable mass and brightness across a wide emission range. This enables the production of nanocrystal-based probes that have comparable reactivity and sensitivity over a large color range. We present spectral imaging results of antibody targeted nanocrystal cluster labeling of target proteins in cultured cells and a Western blot experiment. The combination of spectral imaging with the use of clusters of nanocrystals further improves the sensitivity over either of the approaches independently.

  13. A nucleic acid probe labeled with desmethyl thiazole orange: a new type of hybridization-sensitive fluorescent oligonucleotide for live-cell RNA imaging.

    PubMed

    Okamoto, Akimitsu; Sugizaki, Kaori; Yuki, Mizue; Yanagisawa, Hiroyuki; Ikeda, Shuji; Sueoka, Takuma; Hayashi, Gosuke; Wang, Dan Ohtan

    2013-01-14

    A new fluorescent nucleotide with desmethyl thiazole orange dyes, D'(505), has been developed for expansion of the function of fluorescent probes for live-cell RNA imaging. The nucleoside unit of D'(505) for DNA autosynthesis was soluble in organic solvents, which made the preparation of nucleoside units and the reactions in the cycles of DNA synthesis more efficient. The dyes of D'(505)-containing oligodeoxynucleotide were protonated below pH 7 and the oligodeoxynucleotide exhibited hybridization-sensitive fluorescence emission through the control of excitonic interactions of the dyes of D'(505). The simplified procedure and effective hybridization-sensitive fluorescence emission produced multicolored hybridization-sensitive fluorescent probes, which were useful for live-cell RNA imaging. The acceptor-bleaching method gave us information on RNA in a specific cell among many living cells.

  14. Gaseous environment-sensitive fluorescence of YAG:Ce3+ nanocrystals

    NASA Astrophysics Data System (ADS)

    Feofilov, S. P.; Arsentyev, D. V.; Kulinkin, A. B.; Gacoin, T.; Mialon, G.; Meltzer, R. S.; Dujardin, C.

    2010-03-01

    The influence of the surrounding gaseous media on optically excited fluorescence of YAG:Ce nanoparticles was studied. It was observed that the 5d→4f fluorescence of Ce3+ ions in YAG nanoparticles dramatically depends on the pressure of the gaseous media surrounding the powder samples. The observations suggest the possibility of probing the pressure at the single molecule level with nanoparticles. It is suggested that the observed effects result from gas-molecule-assisted electron transfer between the nanoparticles and from modification of the surface of the particles by adsorption of gas molecules.

  15. A fluorescent graphitic carbon nitride nanosheet biosensor for highly sensitive, label-free detection of alkaline phosphatase

    NASA Astrophysics Data System (ADS)

    Xiang, Mei-Hao; Liu, Jin-Wen; Li, Na; Tang, Hao; Yu, Ru-Qin; Jiang, Jian-Hui

    2016-02-01

    Graphitic C3N4 (g-C3N4) nanosheets provide an attractive option for bioprobes and bioimaging applications. Utilizing highly fluorescent and water-dispersible ultrathin g-C3N4 nanosheets, a highly sensitive, selective and label-free biosensor has been developed for ALP detection for the first time. The developed approach utilizes a natural substrate of ALP in biological systems and thus affords very high catalytic efficiency. This novel biosensor is demonstrated to enable quantitative analysis of ALP in a wide range from 0.1 to 1000 U L-1 with a low detection limit of 0.08 U L-1, which is among the most sensitive assays for ALP. It is expected that the developed method may provide a low-cost, convenient, rapid and highly sensitive platform for ALP-based clinical diagnostics and biomedical applications.Graphitic C3N4 (g-C3N4) nanosheets provide an attractive option for bioprobes and bioimaging applications. Utilizing highly fluorescent and water-dispersible ultrathin g-C3N4 nanosheets, a highly sensitive, selective and label-free biosensor has been developed for ALP detection for the first time. The developed approach utilizes a natural substrate of ALP in biological systems and thus affords very high catalytic efficiency. This novel biosensor is demonstrated to enable quantitative analysis of ALP in a wide range from 0.1 to 1000 U L-1 with a low detection limit of 0.08 U L-1, which is among the most sensitive assays for ALP. It is expected that the developed method may provide a low-cost, convenient, rapid and highly sensitive platform for ALP-based clinical diagnostics and biomedical applications. Electronic supplementary information (ESI) available. See DOI: 10.1039/c5nr08278a

  16. A triazole Schiff base-based selective and sensitive fluorescent probe for Zn2 +: A combined experimental and theoretical study

    NASA Astrophysics Data System (ADS)

    Yuan, Caixia; Liu, Xinyu; Wu, Yanbo; Lu, Liping; Zhu, Miaoli

    2016-02-01

    A triazole-Schiff base, 4-(5-Chloro-2-hydroxybenzylideneamino)-1H-1,2,4-triazole-5(4H)-thione (HL), exhibits the high selectivity and sensitivity for Zn2 + in the fluorescence spectrometry over other common metal ions, especially Cd2 + in DMSO:H2O (1:9, v/v) solution. A 1:1 binding ratio of Zn2 +/L for the complex has been obtained by Uv-Vis titration experiments and Job's plot with the detection limit of 51 nmol/L. The coordination mode of the complex in solution was further confirmed by density functional theory (DFT) calculations. Time-dependent density functional theory (TD-DFT) calculations indicate that a chelation-enhanced fluorescence (CHEF) effect occurs in the process of detecting Zn ion.

  17. A highly selective and sensitive fluorescent sensor for the rapid detection of Hg2 + based on phenylamine-oligothiophene derivative

    NASA Astrophysics Data System (ADS)

    Niu, Qingfen; Wu, Xingxing; Zhang, Shanshan; Li, Tianduo; Cui, Yuezhi; Li, Xiaoyan

    2016-01-01

    A fast-responsive fluorescent phenylamine-oligothiophene sensor 3TDDA was reported. This sensor exhibited highly selective and sensitive detection of Hg2 + ion in aqueous solution (THF/CH3CN/H2O, 45/50/5, v/v) through fluorescence quenching. The detection was not affected by the coexistence of other competitive metal ions such as Na+, K+, Ag+, Ca2 +, Fe3 +, Al3 +, Co2 +, Ni2 +, Zn2 +, Pb2 +, Cd2 +, Fe2 + and Cr3 +. A stoichiometric ratio (1:1) of the sensor and Hg2 + was determined by a Job's plot and mole-ratio curves. The binding of sensor 3TDDA and Hg2 + was also chemically reversible with EDTA. The detection limit was calculated as low as 4.392 × 10- 7 M.

  18. Simple and sensitive high performance liquid chromatography method with fluorescence detection for therapeutic drug monitoring of topiramate.

    PubMed

    Milosheska, Daniela; Vovk, Tomaž; Grabnar, Iztok; Roškar, Robert

    2015-01-01

    A simple, sensitive, accurate, precise and inexpensive HPLC method with fluorescence detection, suitable for routine therapeutic drug monitoring (TDM) of an antiepileptic drug topiramate, was developed and validated. The determination of plasma topiramate concentration was carried out after precolumn derivatization, using 4-chloro-7-nitrobenzofurazan as a fluorescent labeling agent and bendroflumethiazide as an internal standard. The standard calibration curve was linear over the concentration range of 0.01-24 μg/mL (r² > 0.9998). The intra- and inter-day accuracies expressed as bias were from 1.4 to 9.9% and from 1.9 to 10.2%, respectively. The intra- and inter-day precisions were below 7.9% and 2.7%, respectively. The validated method was applied for the measurement of plasma topiramate concentrations in patients with epilepsy. The reported method is appropriate for TDM of topiramate as well as for pharmacokinetic and bioequivalence studies.

  19. A cobalt oxyhydroxide nanoflake-based nanoprobe for the sensitive fluorescence detection of T4 polynucleotide kinase activity and inhibition

    NASA Astrophysics Data System (ADS)

    Cen, Yao; Yang, Yuan; Yu, Ru-Qin; Chen, Ting-Ting; Chu, Xia

    2016-04-01

    Phosphorylation of nucleic acids with 5'-OH termini catalyzed by polynucleotide kinase (PNK) is an inevitable process and has been implicated in many important cellular events. Here, we found for the first time that there was a significant difference in the adsorbent ability of cobalt oxyhydroxide (CoOOH) nanoflakes between single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), which resulted in the fluorescent dye-labeled dsDNA still retaining strong fluorescence emission, while the fluorescence signal of ssDNA was significantly quenched by CoOOH nanoflakes. Based on this discovery, we developed a CoOOH nanoflake-based nanoprobe for the fluorescence sensing of T4 PNK activity and its inhibition by combining it with λ exonuclease cleavage reaction. In the presence of T4 PNK, dye-labeled dsDNA was phosphorylated and then cleaved by λ exonuclease to generate ssDNA, which could adsorb on the CoOOH nanoflakes and whose fluorescence was quenched by CoOOH nanoflakes. Due to the high quenching property of CoOOH nanoflakes as an efficient energy acceptor, a sensitive and selective sensing approach with satisfactory performance for T4 PNK sensing in a complex biological matrix has been successfully constructed and applied to the screening of inhibitors. The developed approach may potentially provide a new platform for further research, clinical diagnosis, and drug discovery of nucleotide kinase related diseases.Phosphorylation of nucleic acids with 5'-OH termini catalyzed by polynucleotide kinase (PNK) is an inevitable process and has been implicated in many important cellular events. Here, we found for the first time that there was a significant difference in the adsorbent ability of cobalt oxyhydroxide (CoOOH) nanoflakes between single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), which resulted in the fluorescent dye-labeled dsDNA still retaining strong fluorescence emission, while the fluorescence signal of ssDNA was significantly quenched by Co

  20. Fluorescent cadmium sulfide nanoparticles for selective and sensitive detection of toxic pesticides in aqueous medium

    NASA Astrophysics Data System (ADS)

    Walia, Shanka; Acharya, Amitabha

    2014-12-01

    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.

  1. Radiochemical Analysis by High Sensitivity Micro X-Ray Fluorescence Detection

    SciTech Connect

    Ning Gao

    2006-05-12

    The primary objective of the project was to develop a novel dual-optic x-ray fluorescence instrument capable of doing radiochemical analysis of high-level radioactive wastes at DOE sites such as Savannah River Site and Hanford Site.

  2. Highly sensitive colorimetric and fluorescent sensor for cyanazine based on the inner filter effect of gold nanoparticles

    NASA Astrophysics Data System (ADS)

    Dong, Liang; Hou, Changjun; Yang, Mei; Fa, Huanbao; Wu, Huixiang; Shen, Caihong; Huo, Danqun

    2016-06-01

    Cyanazine residue poses a great threat to human health and its derivatives would remain in soils, natural waters, and other environmental domains for a long time. Herein, a simple, rapid, and ultra-sensitive analytical method for the determination of cyanazine (CZ) based on inner filter effect (IFE) of Au nanoparticles (AuNPs) on the fluorescence of CdTe quantum dots (QDs) is first described in this study. With the presence of citrate-stabilized AuNPs, the fluorescence of GSH-capped CdTe QDs was remarkably quenched by AuNPs via IFE. The fluorescence of the AuNP-CdTe QD system was recovered upon addition of CZ. CZ can adsorb on to the surface of AuNPs due to its cyano group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the IFE of AuNPs on CdTe QDs was weakened, and the fluorescence intensity of CdTe QDs was recovered accordingly. A good linear correlation for detection of CZ was exhibited from 0.05 to 9 μM, and the detection limit reached 0.1568 μM, which was much lower than the safety limit required by the USA, the UK, and China. In order to probe into the selectivity of AuNPs towards CZ over other pesticides, various frequently used pesticides were mixed with AuNPs. AuNP composite solution shows good selectivity towards CZ among other pesticides. This method was successfully carried out for the assessment of CZ in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost, and non-time-consuming compared with traditional methods.

  3. PROTON MICROPROBE ANALYSIS OF TRACE-ELEMENT VARIATIONS IN VITRINITES IN THE SAME AND DIFFERENT COAL BEDS.

    USGS Publications Warehouse

    Minkin, J.A.; Chao, E.C.T.; Blank, Herma; Dulong, F.T.

    1987-01-01

    The PIXE (proton-induced X-ray emission) microprobe can be used for nondestructive, in-situ analyses of areas as small as those analyzed by the electron microprobe, and has a sensitivity of detection as much as two orders of magnitude better than the electron microprobe. Preliminary studies demonstrated that PIXE provides a capability for quantitative determination of elemental concentrations in individual coal maceral grains with a detection limit of 1-10 ppm for most elements analyzed. Encouraged by the earlier results, we carried out the analyses reported below to examine trace element variations laterally (over a km range) as well as vertically (cm to m) in the I and J coal beds in the Upper Cretaceous Ferron Sandstone Member of the Mancos Shale in central Utah, and to compare the data with the data from two samples of eastern coals of Pennsylvanian age.

  4. Bead-based competitive fluorescence immunoassay for sensitive and rapid diagnosis of cyanotoxin risk in drinking water.

    PubMed

    Yu, Hye-Weon; Jang, Am; Kim, Lan Hee; Kim, Sung-Jo; Kim, In S

    2011-09-15

    Due to the increased occurrence of cyanobacterial blooms and their toxins in drinking water sources, effective management based on a sensitive and rapid analytical method is in high demand for security of safe water sources and environmental human health. Here, a competitive fluorescence immunoassay of microcystin-LR (MCYST-LR) is developed in an attempt to improve the sensitivity, analysis time, and ease-of-manipulation of analysis. To serve this aim, a bead-based suspension assay was introduced based on two major sensing elements: an antibody-conjugated quantum dot (QD) detection probe and an antigen-immobilized magnetic bead (MB) competitor. The assay was composed of three steps: the competitive immunological reaction of QD detection probes against analytes and MB competitors, magnetic separation and washing, and the optical signal generation of QDs. The fluorescence intensity was found to be inversely proportional to the MCYST-LR concentration. Under optimized conditions, the proposed assay performed well for the identification and quantitative analysis of MCYST-LR (within 30 min in the range of 0.42-25 μg/L, with a limit of detection of 0.03 μg/L). It is thus expected that this enhanced assay can contribute both to the sensitive and rapid diagnosis of cyanotoxin risk in drinking water and effective management procedures.

  5. A sensitive fluorescence optosensor for analysing propranolol in pharmaceutical preparations and a test for its control in urine in sport.

    PubMed

    Fernández-Sánchez, J F; Carretero, A Segura; Cruces-Blanco, C; Fernández-Gutiérrez, A

    2003-04-01

    We describe a simple and selective method for analysing propranolol and a sensitive test for its control in urine. A flow-through fluorescence optosensor based on on-line immobilization in a non-ionic-exchanger (Amberlite XAD-7) solid support in a continuous flow was used in both cases. Determination was made in 5 mM H(2)PO(4)(-)/HPO(4)(2-) buffer solution at pH 6 at a working temperature of 20 degrees C. Fluorescence intensities were measured at lambda(ex/em) = 300/338 nm with a response time of 80 s, thus obtaining a linear concentration range of between 0 and 250.0 ng ml(-1) with a detection limit of 1.3 ng ml(-1), an analytical sensitivity of 6.0 ng ml(-1) and a standard deviation of 2.40% at a 150 ng ml(-1) concentration level for propranolol. We also propose a test to detect propranolol in urine with a linear concentration range between 0 and 100.0 ng ml(-1), a detection limit of 0.2 ng ml(-1), an analytical sensitivity of 1.0 ng ml(-1), and a standard deviation of 0.84% at a 75 ng ml(-1) concentration level. The effect of proteins presents in urine samples were evaluated. The two proposed methods were satisfactorily applied to commercial formulations and urine samples respectively.

  6. Ion microscope and ion microprobe analysis under oxygen, cesium and gallium bombardment

    NASA Astrophysics Data System (ADS)

    Migeon, H.-N.; Saldi, F.; Gao, Y.; Schuhmacher, M.

    1995-05-01

    This article concentrates on dynamic SIMS analysis using a magnetic sector instrument at micron and sub-micron resolutions with the ion microscope and ion microprobe modes. The advantages and drawbacks of both alternatives for recording measurements in laterally heterogeneous specimens are highlighted expecially concerning transmission and acquisition times. The ionization efficiencies and matrix effects under oxygen, cesium and gallium bombardment are compared. The ion microscope is shown to provide fast acquisition times owing to the parallel detection of the entire analyzed area and the most adequate mode for lateral resolutions above 1 [mu]m, whereas the ion microprobe provides better sensitivity and is best suited for high resolution. Combining cesium and oxygen ion sources provides, in most cases, a better ionization efficiency than the gallium beam but all three sources induce matrix effects which are shown to be much less critical using cationized species.

  7. A metal-free turn-on fluorescent probe for the fast and sensitive detection of inorganic azides.

    PubMed

    Wang, Ke; Friscourt, Frédéric; Dai, Chaofeng; Wang, Lifang; Zheng, Yueqin; Boons, Geert-Jan; Wang, Siming; Wang, Binghe

    2016-04-01

    Sodium azide is toxic and widely used in agricultural, commercial products, and research laboratories. Thus it is of a significant environmental concern and there is a need for the development of a rapid detection method. A fluorogenic dibenzylcyclooctyne derivative (Fl-DIBO) is herein described as a fluorescent probe for the rapid detection of inorganic azide via Strain-Promoted Azide-Alkyne Cycloaddition (SPAAC). Fl-DIBO was found to be highly selective toward NaN3 in comparison to other common anions with good sensitivity and detection limit of 10μM.

  8. Highly multiplex and sensitive SNP genotyping method using a three-color fluorescence-labeled ligase detection reaction coupled with conformation-sensitive CE.

    PubMed

    Choi, Woong; Jung, Gyoo Yeol

    2017-02-01

    For the development of clinically useful genotyping methods for SNPs, accuracy, simplicity, sensitivity, and cost-effectiveness are the most important criteria. Among the methods currently being developed for SNP genotyping technology, the ligation-dependent method is considered the simplest for clinical diagnosis. However, sensitivity is not guaranteed by the ligation reaction alone, and analysis of multiple targets is limited by the detection method. Although CE is an attractive alternative to error-prone hybridization-based detection, the multiplex assay process is complicated because of the size-based DNA separation principle. In this study, we employed the ligase detection reaction coupled with high-resolution CE-SSCP to develop an accurate, sensitive, and simple multiplex genotyping method. Ligase detection reaction could amplify ligated products through recurrence of denaturation and ligation reaction, and SSCP could separate these products according to each different structure conformation without size variation. Thus, simple and sensitive SNP analysis can be performed using this method involving the use of similar-sized probes, without complex probe design steps. We found that this method could not only accurately discriminate base mismatches but also quantitatively detect 37 SNPs of the tp53 gene, which are used as targets in multiplex analysis, using three-color fluorescence-labeled probes.

  9. Endoscopic fluorescence of gastrointestinal neoplasia after sensitization with 5-aminolaevulinic acid (ALA) or Photofrin

    NASA Astrophysics Data System (ADS)

    Messmann, Helmut; Mlkvy, Peter; Montan, Sune; Wang-Nordman, Ingrid; Nilsson, Annika M.; Svanberg, Katarina; Svanberg, Sune; MacRobert, Alexander J.; Bown, Stephen G.

    1995-03-01

    Fluorescence after photosensitization has the potential to identify lesions not visible on conventional endoscopy. We assessed 12 patients at high risk of or with established GI cancers (u ulcerative colitis, 1 colon polyp, 2 familial polyposis with duodenal polyps, 2 early oesophageal cancers). Fluorescence images (excitation 390 nm) were recorded with endoscopic equipment and additional spot measurements (optical multichannel analyzer). Patients were given 10 - 60 mg/kg ALA orally or 2 mg/kg Photofrin i.v. 60 mg/kg ALA gave high levels of PP IX (proto-porphyrin IX) in all areas, but 10 - 15 mg/kg resulted in selectivity in macroscopically inflamed colon. Photofrin gave oesophageal tumors selectivity at 4 and 48 hours. Photofrin patients subsequently had PDT. Photobleaching was documented in 3. We conclude that these techniques have potential as `optical biopsy tools' and for screening for early neoplastic changes.

  10. An environmentally sensitive fluorescent purine nucleoside that changes emission wavelength upon hybridization.

    PubMed

    Saito, Yoshio; Suzuki, Azusa; Okada, Yuji; Yamasaka, Yuki; Nemoto, Nobukatsu; Saito, Isao

    2013-06-25

    C7-naphthylethynylated 8-aza-7-deaza-2'-deoxyguanosine (na)G was synthesized and its photophysical properties were examined. The fluorescent nucleoside exhibited solvatofluorochromic properties (Δλ(fl)(max) = 67 nm). An ODN probe containing (na)G forms a stable base pair only with C and discriminates structural changes such as mismatches and deletions by a distinct change in its emission wavelength.

  11. In situ measurement of airway surface liquid [K+] using a ratioable K+-sensitive fluorescent dye.

    PubMed

    Namkung, Wan; Song, Yuanlin; Mills, Aaron D; Padmawar, Prashant; Finkbeiner, Walter E; Verkman, A S

    2009-06-05

    The airway surface liquid (ASL) is the thin fluid layer lining airway surface epithelial cells, whose volume and composition are tightly regulated and may be abnormal in cystic fibrosis (CF). We synthesized a two-color fluorescent dextran to measure ASL [K(+)], TAC-Lime-dextran-TMR, consisting of a green-fluorescing triazacryptand K(+) ionophore-Bodipy conjugate, coupled to dextran, together with a red fluorescing tetramethylrhodamine reference chromophore. TAC-Lime-dextran-TMR fluorescence was K(+)-selective, increasing >4-fold with increasing [K(+)] from 0 to 40 mm. In well differentiated human airway epithelial cells, ASL [K(+)] was 20.8 +/- 0.3 mm and decreased by inhibition of the Na(+)/K(+) pump (ouabain), ENaC (amiloride), CF transmembrane conductance regulator (CFTR(inh)-172), or K(+) channels (TEA or XE991). ASL [K(+)] was increased by forskolin but not affected by Na(+)/K(+)/2Cl(-) cotransporter inhibition (bumetanide). Functional and expression studies indicated the involvement of [K(+)] channels KCNQ1, KCNQ3, and KCNQ5 as determinants of ASL [K(+)]. [K(+)] in CF cultures was similar to that in non-CF cultures, suggesting that abnormal ASL [K(+)] is not a factor in CF lung disease. In intact airways, ASL [K(+)] was also well above extracellular [K(+)]: 22 +/- 1 mm in pig trachea ex vivo and 16 +/- 1 mm in mouse trachea in vivo. Our results provide the first noninvasive measurements of [K(+)] in the ASL and indicate the involvement of apical and basolateral membrane ion transporters in maintaining a high ASL [K(+)].

  12. Supramolecular interaction of labetalol with cucurbit[7]uril for its sensitive fluorescence detection.

    PubMed

    Li, Changfeng; Feng, Jianxia; Ju, Huangxian

    2015-01-07

    This work studied the host-guest interaction between cucurbit[7]uril (CB[7]) and labetalol in acidic aqueous solution and proposed a simple competitive method for fluorescence detection of labetalol. The binding constant of labetalol-CB[7] was (1.83 ± 0.22) × 10(6) M(-1), which was greater than those of palmatine-CB[7], berberine-CB[7], and coptisine-CB[7] complexes. The fluorescence intensity of palmatine-CB[7], berberine-CB[7], and coptisine-CB[7] complexes decreased linearly with increasing concentration of labetalol ranging from 0.014 to 2.06, 0.014 to 1.15, and 0.034 to 1.23 μM, respectively. Based on the competitive interaction, the proposed detection method for labetalol showed limits of detection of 4.9 nM, 4.9 nM, and 12.0 nM, respectively, and was successfully applied for the determination of labetalol in human urine samples with good precision and recoveries from 95.4% to 102.5%. Moreover, it could be employed to monitor the time-dependent concentration of labetalol in urine from a healthy volunteer after oral medication. The superstructure-based competitive mode provided a promising fluorescence assay strategy for various potential applications.

  13. Chromatic aberration correction and deconvolution for UV sensitive imaging of fluorescent sterols in cytoplasmic lipid droplets.

    PubMed

    Wüstner, Daniel; Faergeman, Nils J

    2008-08-01

    Intrinsically fluorescent sterols, like dehydroergosterol (DHE), mimic cholesterol closely and are therefore suitable to determine cholesterol transport by fluorescence microscopy. Disadvantages of DHE are its low quantum yield, rapid bleaching, and the fact that its excitation and emission is in the UV region of the spectrum. Thus, one has to deal with chromatic aberration and low signal-to-noise ratio. We developed a method to correct for chromatic aberration between the UV channel and the red/green channel in multicolor imaging of DHE compared with the lipid droplet marker Nile Red in living macrophage foam cells and in adipocytes. We used deconvolution microscopy and developed image segmentation techniques to assess the DHE content of lipid droplets in both cell types in an automated manner. Pulse-chase studies and colocalization analysis were performed to monitor the redistribution of DHE upon adipocyte differentiation. DHE is targeted to transferrin-positive recycling endosomes in preadipocytes but associates with droplets in mature adipocytes. Only in adipocytes but not in foam cells fluorescent sterol was confined to the droplet-limiting membrane. We developed an approach to visualize and quantify sterol content of lipid droplets in living cells with potential for automated high content screening of cellular sterol transport.

  14. Aerothermal Heating Predictions for Mars Microprobe

    NASA Technical Reports Server (NTRS)

    Mitcheltree, R. A.; DiFulvio, M.; Horvath, T. J.; Braun, R. D.

    1998-01-01

    A combination of computational predictions and experimental measurements of the aerothermal heating expected on the two Mars Microprobes during their entry to Mars are presented. The maximum, non-ablating, heating rate at the vehicle's stagnation point (at alpha = 0 degrees) is predicted for an undershoot trajectory to be 194 Watts per square centimeters with associated stagnation point pressure of 0.064 atm. Maximum stagnation point pressure occurs later during the undershoot trajectory and is 0.094 atm. From computations at seven overshoot-trajectory points, the maximum heat load expected at the stagnation point is near 8800 Joules per square centimeter. Heat rates and heat loads on the vehicle's afterbody are much lower than the forebody. At zero degree angle-of-attack, heating over much of the hemi-spherical afterbody is predicted to be less than 2 percent of the stagnation point value. Good qualitative agreement is demonstrated for forebody and afterbody heating between CFD calculations at Mars entry conditions and experimental thermographic phosphor measurements from the Langley 20-Inch Mach 6 Air Tunnel. A novel approach which incorporates six degree-of-freedom trajectory simulations to perform a statistical estimate of the effect of angle-of-attack, and other off-nominal conditions, on heating is included.

  15. Stand-alone microprobe at Livermore

    SciTech Connect

    Antolak, A J; Bench, G S; Brown, T A; Frantz, B R; Grant, P G; Morse, D H; Roberts, M L

    1998-10-02

    Lawrence Livermore National Laboratory (LLNL) and Sandia National Laboratories/California have jointly constructed a new stand-alone microprobe facility. Although the facility was built to develop a method to rapidly locate and determine elemental concentrations of micron scale particulates on various media using PIXE, the facility has found numerous applications in biology and materials science. The facility is located at LLNL and uses a General Ionex Corporation Model 358 duoplasmatron negative ion source, a National Electrostatics Corporation 5SDH-2 tandem accelerator, and an Oxford triplet lens. Features of the system include complete computer control of the beam transport using LabVIEWTM for Macintosh, computer controlled beam collimating and divergence limiting slits, automated sample positioning to micron resolution, and video optics for beam positioning and sample observation. Data collection is accomplished with the simultaneous use of as many as four EG&G Ortec IGLET-XTM X-Ray detectors, digital amplifiers made by X-Ray Instruments and Associates (XIA), and LabVIEWTM for Macintosh acquisition software.

  16. The new nuclear microprobe at Livermore

    NASA Astrophysics Data System (ADS)

    Roberts, M. L.; Bench, G. S.; Heikkinen, D. W.; Morse, D. H.; Bach, P. R.; Pontau, A. E.

    1995-09-01

    Lawrence Livermore National Laboratory (LLNL) and Sandia National Laboratories/California have jointly constructed a new nuclear microprobe beamline. This beamline is located on the LLNL 10 MV tandem accelerator and can be used for multidisciplinary research using PIXE, PIGE, energy loss tomography, or IBS techniques. Distinctive features of the beamline include incorporation of magnet power supplies into the accelerator control system, computer-controlled object and image slits, automated target positioning to sub-micron resolution, and video optics for beam positioning and observation. Mitigation of vibrations was accomplished with vibration isolators and a rigid beamline design while integral beamline shielding was used to shield from stray magnetic fields. Available detectors include a wavelength dispersive X-ray spectrometer, a High-Purity Germanium detector (HPGe), a Lithium-Drifted Silicon X-Ray detector (SiLi), and solid state surface barrier detectors. Along with beamline performance, results from recent measurements on determination of trace impurities in an International Thermonuclear Experimental Reactor (ITER) super conducting wire strand, determination of Ca/Sr ratios in seashells, and determination of minor and trace element concentrations in sperm cells are presented.

  17. Deep Space 2: The Mars Microprobe Mission

    NASA Astrophysics Data System (ADS)

    Smrekar, Suzanne; Catling, David; Lorenz, Ralph; Magalhães, Julio; Moersch, Jeffrey; Morgan, Paul; Murray, Bruce; Presley-Holloway, Marsha; Yen, Albert; Zent, Aaron; Blaney, Diana

    The Mars Microprobe Mission will be the second of the New Millennium Program's technology development missions to planetary bodies. The mission consists of two penetrators that weigh 2.4 kg each and are being carried as a piggyback payload on the Mars Polar Lander cruise ring. The spacecraft arrive at Mars on December 3, 1999. The two identical penetrators will impact the surface at ~190 m/s and penetrate up to 0.6 m. They will land within 1 to 10 km of each other and ~50 km from the Polar Lander on the south polar layered terrain. The primary objective of the mission is to demonstrate technologies that will enable future science missions and, in particular, network science missions. A secondary goal is to acquire science data. A subsurface evolved water experiment and a thermal conductivity experiment will estimate the water content and thermal properties of the regolith. The atmospheric density, pressure, and temperature will be derived using descent deceleration data. Impact accelerometer data will be used to determine the depth of penetration, the hardness of the regolith, and the presence or absence of 10 cm scale layers.

  18. A fluorescence high throughput screening method for the detection of reactive electrophiles as potential skin sensitizers

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Skin sensitization is an important toxicological end-point in the risk assessment of chemical allergens. Because of the complexity of the biological mechanisms associated with skin sensitization integrated approaches combining different chemical, biological and in silico methods are recommended to r...

  19. Development of a Tender-Energy Microprobe for Geosciences at NSLS and NSLS-II

    SciTech Connect

    Northrup, Paul A.

    2014-08-30

    This funding is to develop a new Synchrotron user facility for microbeam X-ray absorption spectroscopy (XAS) and quantitative X-ray fluorescence (XRF) imaging, at the National Synchrotron Light Source (NSLS) and NSLS-II. It includes design, purchase of components, and construction of the microprobe endstation and controls. Initial development, commissioning, and application is ongoing at NSLS Beamline X15B, with planned transition in 2014-15 to the NSLS-II TES (Tender-Energy Spatially Resolved X-ray Absorption Spectroscopy) beamline. It is optimized for the “tender” energy range of 1-5 keV, reaching up to 8 keV. Thus it uniquely covers the K absorption edges of critical elements Mg, Al, Si, P, S, Cl, and Ca, and can reach up to Co. A stable, high-flux microbeam focus, user-tunable from ~50 to ~5 microns, has been achieved using two-stage achromatic focusing. Existing beamline optics collimate, monochromate, and macro-focus the X-ray beam to ~1 mm at a secondary source aperture (SSA). Beam from the SSA is then re-focused by a pair of mirrors in KB geometry to the microbeam scale. Size of the microbeam is tunable, at the expense of flux, by adjusting the size of the SSA as a virtual source. The new experimental endstation consists of 1) a sample chamber operable as a radiation enclosure with helium atmosphere to facilitate measurements in this energy range, 2) the KB microfocusing optics, 3) a sample-positioning stage for raster-scanning and positioning the sample, 4) X-ray fluorescence detectors, an existing Ge detector for low-signal sensitivity and a new Si detector for high count rates, 5) an optical camera for viewing samples and locating target locations, 6) beam intensity monitors and diagnostics, and 7) controls and data acquisition system. An important aspect of this project is the added capability for fast, on-the-fly scanning of the monochromator (energy), required for fast XAS and advanced XAS imaging. This instrument will be available for initial PI

  20. A highly sensitive fluorescent chemosensor for selective detection of zinc (II) ion based on the oxadiazole derivative

    NASA Astrophysics Data System (ADS)

    Lin, Lu; Wang, Dan; Chen, Si-Hong; Wang, Dun-Jia; Yin, Guo-Dong

    2017-03-01

    A novel fluorescent chemosensor based on the oxadiazole, 2-(2-hydroxyphenyl)-5-(4-methoxyphenyl)-1,3,4-oxadiazole, was designed and synthesized. The interaction of the oxadiazole with different metal ions had been investigated through UV-vis absorption and fluorescence spectra in 9:1 (v/v) ethanol-water (pH = 7.0) solution. The oxadiazole showed a pronounced fluorescence enhancement at 430 nm upon addition of Zn2 + in aqueous solution, whereas it had no apparent interference from other metal ions. The results indicated that the oxadiazole possessed high selectivity and sensitivity to Zn2 + ion. The stoichiometric ratio between the oxadiazole and Zn2 + ion was calculated to be 2:1 by Job plot experiment, meanwhile their binding modes was confirmed by 1H NMR and mass spectrometry. Their association constant was determined to be 1.95 × 105 M- 1 and the detection limit for Zn2 + ion was 6.14 × 10- 7 mol/L.

  1. Micron-sized surface enhanced Raman scattering reporter/fluorescence probe encoded colloidal microspheres for sensitive DNA detection.

    PubMed

    You, Lijun; Li, Ruimin; Dong, Xu; Wang, Fang; Guo, Jia; Wang, Changchun

    2017-02-15

    A new type of optical probes, featuring surface enhanced Raman scattering (SERS) and fluorescence spectra dual-mode encoding, has been reported in this article. Based on the uniform micrometer-sized melamine resin/Ag nanoparticles (MRM/Ag-NPs) composite microspheres, the SERS reporters and fluorescent probes were successfully fixed onto the different layers of the MEM/Ag-NPs microspheres, which supported the sensitive DNA detecton. The two spectroscopic methods commonly considered to be contradictive to each other, yet the optical signals were separable in the experiments. The dual-encoding strategy and single microsphere detecton method put the number of available independent codes to be rough the multiple of those available in the two optical detection channels, which increases far more rapidly than the summation of the two channels. As a proof of cencept, the utility of this dual spectrum mode SERS-fluoresecence encoded microsphere (SFEM) was demonstrated in a specific DNA detection using complimentary ssDNA functionalized magnetic beads as the DNA capturing and separation agents. Excellent encoding results were demonstrated from the decoding of the SERS and fluorescence signals of the SFEM. The method appears to be general in scope and we expect that the SERS-fluoresecence encoded microspheres system is applicable to multiplex bioassays of a variety of biomolecules.

  2. The inhibition of fluorescence resonance energy transfer between multicolor quantum dots for rapid and sensitive detection of Staphylococcus aureus

    NASA Astrophysics Data System (ADS)

    Wang, Beibei; Wang, Qi; Ma, Meihu; Cai, Zhaoxia

    2015-01-01

    In this paper, we constructed the fluorescence resonance energy transfer (FRET) system between two multi-color quantum dots (QDs) of two sizes for rapid and sensitive detection of Staphylococcus aureus. In this system, green-emitting QDs conjugated with rabbit anti-S. aureus antibodies were used as energy donors while orange-emitting QDs conjugated with goat-anti-rabbit IgG were used as energy acceptors to form FRET system. Pre-binding of Staphylococcus aureus (S. aureus) on the donor occupied the binding sites and thus blocked resonance energy transfer between two colors QDs, leading to the quenching fluorescence of the acceptor. The fluorescence of acceptor has a linear calibration graph with the concentrations of S. aureus from 52 to 2.6 × 105 CFU mL-1. The low detection limit was 10 CFU/mL. It was worth mentioning that the detection method of S. aureus had been applied to the analysis of apple juice and milk samples, which could potentially be developed into a sensor in the further study.

  3. Silica-coated ZnS quantum dots as fluorescent probes for the sensitive detection of Pb2+ ions

    NASA Astrophysics Data System (ADS)

    Qu, Hua; Cao, Lixin; Su, Ge; Liu, Wei; Gao, Rongjie; Xia, Chenghui; Qin, Junjie

    2014-12-01

    The silica-coated ZnS quantum dots (ZnS@SiO2 QDs) were prepared via a simple and environmentally friendly process. The oil-soluble ZnS cores were successfully transferred to water by the coating of SiO2 shells. The QDs exhibited satisfying dispersion and luminescent properties in water. The ZnS@SiO2 QDs were directly used as fluorescent probes for heavy metal ions without the addition of any buffer solution. The luminescence of QDs was extremely sensitive to Pb2+ ions, and the fluorescence quenching was well described by the Stern-Volmer equation, with an even quenching constant for the Pb2+ ions samples concentration ranging from 10-9 to 2.6 × 10-4 M. An extended hypothesis based on the traditional cation exchange mechanism is proposed to analyze the most significant fluorescence quenching effect by Pb2+ ions. Studies show that ZnS@SiO2 QDs have great potentials to be a sensor for Pb2+ analysis at low to high concentrations.

  4. Heterogeneous nano metal-organic framework fluorescence probe for highly selective and sensitive detection of hydrogen sulfide in living cells.

    PubMed

    Ma, Yu; Su, Hao; Kuang, Xuan; Li, Xiangyuan; Zhang, Tingting; Tang, Bo

    2014-11-18

    Hydrogen sulfide (H2S) has been regarded as the third important gaseous signaling molecule involved in human physiological and pathological processes. Due to the high reactive and diffusible properties of H2S, real-time detection of H2S fluctuations in living biological specimens is crucial. Here, we present a Cu(II)-metalated 3D porous nanoscale metal-organic framework (nano-MOF) {CuL[AlOH]2}n (PAC; H6L = meso-tetrakis(4-carboxylphenyl)porphyrin) and successfully employ this nano-MOF as a novel heterogeneous fluorescence probe for H2S detection. As far as we know, nano-MOFs have never been used as selective fluorescence probes for H2S detection. On the basis of the advantages of nano-MOF materials, this biocompatible nano-MOF probe exhibits rapid response, excellent selectivity, and hypotoxicity in in situ detection of H2S and represents the most sensitive fluorescence probe for selective H2S detection under physiological pH. In addition, confocal imaging was achieved successfully in living cells.

  5. A simple and sensitive fluorescent sensor for methyl parathion based on L-tyrosine methyl ester functionalized carbon dots.

    PubMed

    Hou, Juying; Dong, Jing; Zhu, Haishuang; Teng, Xue; Ai, Shiyun; Mang, Minglin

    2015-06-15

    In this paper, a simple and sensitive fluorescent sensor for methyl parathion is developed based on L-tyrosine methyl ester functionalized carbon dots (Tyr-CDs) and tyrosinase system. The carbon dots are obtained by simple hydrothermal reaction using citric acid as carbon resource and L-tyrosine methyl ester as modification reagent. The carbon dots are characterized by transmission electron microscope, high resolution transmission electron microscopy, X-ray diffraction spectrum, Fourier transform infrared spectroscopy, and X-ray photoelectron spectroscopy. The carbon dots show strong and stable photoluminescence with a quantum yield of 3.8%. Tyrosinase can catalyze the oxidation of tyrosine methyl ester on the surface of carbon dots to corresponding quinone products, which can quench the fluorescence of carbon dots. When organophosphorus pesticides (OPs) are introduced in system, they can decrease the enzyme activity, thus decrease the fluorescence quenching rate. Methyl parathion, as a model of OPs, was detected. Experimental results show that the enzyme inhibition rate is proportional to the logarithm of the methyl parathion concentration in the range 1.0×10(-10)-1.0×10(-4) M with the detection limit (S/N=3) of 4.8×10(-11) M. This determination method shows a low detection limit, wide linear range, good selectivity and high reproducibility. This sensing system has been successfully used for the analysis of cabbage, milk and fruit juice samples.

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

    PubMed Central

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

    2013-01-01

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

  7. Boronic acid functionalized N-doped carbon quantum dots as fluorescent probe for selective and sensitive glucose determination

    NASA Astrophysics Data System (ADS)

    Jiang, Guohua; Jiang, Tengteng; Li, Xia; Wei, Zheng; Du, Xiangxiang; Wang, Xiaohong

    2014-04-01

    Nitrogen doped carbon quantum dots (NCQDs) of about 10 nm in diameter have been obtained by hydrothermal reaction from collagen. Because of the superiority of water dispersion, low toxicity and ease of functionlization, the NCQDs were designed as a glucose sensor after covalent grafting by 3-aminophenylboronic (APBA) (APBA-NCQDs). The as-prepared APBA-NCQDs were imparted with glucose sensitivity and selectivity from other saccharides via fluorescence (FL) quenching effect at physiological pH and at room temperature, which show high sensitivity and specificity for glucose determination with a wide range from 1 mM to 14 mM. FL quenching mechanism of APBA-NCQDs was also investigated by adding an external quencher. The APBA-NCQDs-based platform is an environmentally friendly way to substitute inorganic quantum dots containing heavy metals which offer a facile and low cost detection method.

  8. Hierarchical self-assembly of switchable nucleolipid supramolecular gels based on environmentally-sensitive fluorescent nucleoside analogs

    NASA Astrophysics Data System (ADS)

    Nuthanakanti, Ashok; Srivatsan, Seergazhi G.

    2016-02-01

    Exquisite recognition and folding properties have rendered nucleic acids as useful supramolecular synthons for the construction of programmable architectures. Despite their proven applications in nanotechnology, scalability and fabrication of nucleic acid nanostructures still remain a challenge. Here, we describe a novel design strategy to construct new supramolecular nucleolipid synthons by using environmentally-sensitive fluorescent nucleoside analogs, based on 5-(benzofuran-2-yl)uracil and 5-(benzo[b]thiophen-2-yl)uracil cores, as the head group and fatty acids, attached to the ribose sugar, as the lipophilic group. These modified nucleoside-lipid hybrids formed organogels driven by hierarchical structures such as fibers, twisted ribbons, helical ribbons and nanotubes, which depended on the nature of fatty acid chain and nucleobase modification. NMR, single crystal X-ray and powder X-ray diffraction studies revealed the coordinated interplay of various non-covalent interactions invoked by modified nucleobase, sugar and fatty acid chains in setting up the pathway for the gelation process. Importantly, these nucleolipid gels retained or displayed aggregation-induced enhanced emission and their gelation behavior and photophysical properties could be reversibly switched by external stimuli such as temperature, ultrasound and chemicals. Furthermore, the switchable nature of nucleolipid gels to chemical stimuli enabled the selective two channel recognition of fluoride and Hg2+ ions through visual phase transition and fluorescence change. Fluorescent organogels exhibiting such a combination of useful features is rare, and hence, we expect that this innovative design of fluorescent nucleolipid supramolecular synthons could lead to the emergence of a new family of smart optical materials and probes.Exquisite recognition and folding properties have rendered nucleic acids as useful supramolecular synthons for the construction of programmable architectures. Despite their

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

    PubMed

    Zhang, Dapeng; Yin, Lei; Meng, Zihui; Yu, Anchi; Guo, Lianghong; Wang, Hailin

    2014-02-17

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

  10. Magnetic-fluorescent-targeting multifunctional aptasensorfor highly sensitive and one-step rapid detection of ochratoxin A.

    PubMed

    Wang, Chengquan; Qian, Jing; Wang, Kan; Wang, Kun; Liu, Qian; Dong, Xiaoya; Wang, Chengke; Huang, Xingyi

    2015-06-15

    A multifunctional aptasensor for highly sensitive and one-step rapid detection of ochratoxin A (OTA), has been developed using aptamer-conjugated magnetic beads (MBs) as the recognition and concentration element and a heavy CdTe quantum dots (QDs) as the label. Initially, the thiolated aptamer was conjugated on the Fe3O4@Au MBs through Au-S covalent binding. Subsequently, multiple CdTe QDs were loaded both in and on a versatile SiO2 nanocarrier to produce a large amplification factor of hybrid fluorescent nanoparticles (HFNPs) labeled complementary DNA (cDNA). The magnetic-fluorescent-targeting multifunctional aptasensor was thus fabricated by immobilizing the HFNPs onto MBs' surface through the hybrid reaction between the aptamer and cDNA. This aptasensor can be produced at large scale in a single run, and then can be conveniently used for rapid detection of OTA through a one-step incubation procedure. The presence of OTA would trigger aptamer-OTA binding, resulting in the partial release of the HFNPs into bulk solution. After a simple magnetic separation, the supernatant liquid of the above solution contained a great number of CdTe QDs produced an intense fluorescence emission. Under the optimal conditions, the fluorescence intensity of the released HFNPs was proportional to the concentration of OTA in a wide range of 15 pg mL(-1) -100 ng mL(-1) with a detection limit of 5.4 pg mL(-1) (S/N=3). This multifunctional aptasensor represents a promising path toward routine quality control of food safety, and also creates the opportunity to develop aptasensors for other targets using this strategy.

  11. Ultrasound-Triggered Phase Transition Sensitive Magnetic Fluorescent Nanodroplets as a Multimodal Imaging Contrast Agent in Rat and Mouse Model

    PubMed Central

    Chen, Yunchao; Luo, Binhua; Liu, Xuhan; Liu, Wei; Xu, Haibo; Yang, Xiangliang

    2013-01-01

    Ultrasound-triggered phase transition sensitive nanodroplets with multimodal imaging functionality were prepared via premix Shirasu porous glass (SPG) membrane emulsification method. The nanodroplets with fluorescence dye DiR and SPIO nanoparticles (DiR-SPIO-NDs) had a polymer shell and a liquid perfluoropentane (PFP) core. The as-formed DiR-SPIO-NDs have a uniform size of 385±5.0 nm with PDI of 0.169±0.011. The TEM and microscopy imaging showed that the DiR-SPIO-NDs existed as core-shell spheres, and DiR and SPIO nanoparticles dispersed in the shell or core. The MTT and hemolysis studies demonstrated that the nanodroplets were biocompatible and safe. Moreover, the proposed nanodroplets exhibited significant ultrasound-triggered phase transition property under clinical diagnostic ultrasound irradiation due to the vaporization of PFP inside. Meanwhile, the high stability and R2 relaxivity of the DiR-SPIO-NDs suggested its applicability in MRI. The in vivo T2-weighted images of MRI and fluorescence images both showed that the image contrast in liver and spleen of rats and mice model were enhanced after the intravenous injection of DiR-SPIO-NDs. Furthermore, the ultrasound imaging (US) in mice tumor as well as MRI and fluorescence imaging in liver of rats and mice showed that the DiR-SPIO-NDs had long-lasting contrast ability in vivo. These in vitro and in vivo findings suggested that DiR-SPIO-NDs could potentially be a great MRI/US/fluorescence multimodal imaging contrast agent in the diagnosis of liver tissue diseases. PMID:24391983

  12. Hierarchical self-assembly of switchable nucleolipid supramolecular gels based on environmentally-sensitive fluorescent nucleoside analogs.

    PubMed

    Nuthanakanti, Ashok; Srivatsan, Seergazhi G

    2016-02-14

    Exquisite recognition and folding properties have rendered nucleic acids as useful supramolecular synthons for the construction of programmable architectures. Despite their proven applications in nanotechnology, scalability and fabrication of nucleic acid nanostructures still remain a challenge. Here, we describe a novel design strategy to construct new supramolecular nucleolipid synthons by using environmentally-sensitive fluorescent nucleoside analogs, based on 5-(benzofuran-2-yl)uracil and 5-(benzo[b]thiophen-2-yl)uracil cores, as the head group and fatty acids, attached to the ribose sugar, as the lipophilic group. These modified nucleoside-lipid hybrids formed organogels driven by hierarchical structures such as fibers, twisted ribbons, helical ribbons and nanotubes, which depended on the nature of fatty acid chain and nucleobase modification. NMR, single crystal X-ray and powder X-ray diffraction studies revealed the coordinated interplay of various non-covalent interactions invoked by modified nucleobase, sugar and fatty acid chains in setting up the pathway for the gelation process. Importantly, these nucleolipid gels retained or displayed aggregation-induced enhanced emission and their gelation behavior and photophysical properties could be reversibly switched by external stimuli such as temperature, ultrasound and chemicals. Furthermore, the switchable nature of nucleolipid gels to chemical stimuli enabled the selective two channel recognition of fluoride and Hg(2+) ions through visual phase transition and fluorescence change. Fluorescent organogels exhibiting such a combination of useful features is rare, and hence, we expect that this innovative design of fluorescent nucleolipid supramolecular synthons could lead to the emergence of a new family of smart optical materials and probes.

  13. Biological Effects in Coral Biomineralization: The Ion-Microprobe Revolution

    NASA Astrophysics Data System (ADS)

    Meibom, A.

    2004-12-01

    Scleractinian corals are among the most prolific biomineralizing organisms on Earth and massive, reef-building corals are used extensively as proxies for past variations in the global climate. It is therefore of wide interest to understand the degree to which biological versus inorganic processes control the chemistry of the coral skeleton. Early workers considered aragonitic coral skeleton formation to be a purely physiochemical process. More recent studies have increasingly emphasized the role of a skeletal organic matrix, or intercalated organic macro-molecules that control the macroscopic shape and size of the growing crystals. It is now well established that organic compounds play a key role in controlling the morphology of crystals in a wide variety of calcium carbonate biomineralization processes by binding to specific sites, thereby causing direction-specific binding energies on the crystal surfaces. Macro-molecules, such as aspartic acid-rich or glutamic proteins and sulfated polysaccharides, are known to be embedded within the aragonitic skeletal components of coral. In addition, endosymbiotic algae and the layer of cells adjacent to the mineralizing surface, the calicoblastic ectoderm, are believed to play important roles in driving and controlling hermatypic coral skeletogenesis. However, until recently, further progress has been somewhat limited because it was not possible to obtain chemical analyses of the coral skeleton with sufficiently high spatial resolution and sensitivity to correlate chemical variations with the micrometer scale organization of its different structural components. The recent emergence of new ion microprobe technology is changing this situation radically. Conventional ion microprobe and laser ablation techniques have already contributed substantially to our knowledge about the micro-distribution of key trace elements such as B, Mg, Sr, Ba and U. However, with the development of the NanoSIMS, a newly designed ion microprobe

  14. The Perils of Electron Microprobe Analysis of Apatite

    NASA Astrophysics Data System (ADS)

    Henderson, C. E.; Essene, E. J.; Wang, K. L.; Zhang, Y.

    2010-12-01

    . Infrared spectra show a strong band of (CO3)2- for this apatite, which indicates a possible substitution of (CO3)2-(F)- for (PO4)3-. Other techniques to mitigate temporal variation of F and Cl, including alternative metal coatings, concurrent stage movement, and cryogenic sample-cooling were attempted, but did not eliminate the disparity in measured F concentrations between the two sample orientations. Thus, we believe that F measurements on F-rich apatite samples of unknown orientation are immediately suspect and should be regarded as upper limits of true F concentration. X-ray mapping, CL imaging and subsequent quantitative analyses show compositional variations in Na, S, Si, and REE in the Durango and Wilberforce fluorapatite samples used in this study. Problems of electron beam sensitivity, X-ray intensity anisotropy due to sample orientation, and compositional heterogeneity call into question their continued use as routine microanalysis reference materials. Microanalysts are encouraged to use more robust calibration standards, such as Cl-rich or other F-poor apatites for Ca, P, O and Cl, and MgF2 for F measurements. [1] Stormer, J.C., Pierson, M.L, and Tacker, R.C. (1993) Variation of F and Cl X-ray intensity due to anisotropic diffusion in apatite during electron microprobe analysis. Am. Min., 78, 641-648.

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

    NASA Astrophysics Data System (ADS)

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

    1998-04-01

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

  16. Ultra sensitive sensor with enhanced dynamic range for high speed detection of multi-color fluorescence radiation.

    PubMed

    Tsupryk, A; Tovkach, I; Gavrilov, D; Kosobokova, O; Gudkov, G; Tyshko, G; Gorbovitski, B; Gorfinkel, V

    2008-05-15

    This paper describes design of the new ultra sensitive sensor system for fluorescence detection applications. System comprises two units: optical spectra separation unit and detection unit. Optical unit of the sensor performs spatial spectra separation of signal from the laser excited fluorescence, and resulting spectra is collected in the detection part of the system. Optical part is built using diffraction grating as spectra separation element. Detection part comprises 32-channel photomultiplier tube working in single photon counting mode with our 32-channel amplifier. Using single photon detection technique and specific signal processing algorithms for collected data, the proposed system allows to achieve unique combination of characteristics--high sensitivity, high detection speed and wide linearity dynamic range comparing to existing commercial instruments. DNA sequencing experiments with new sensor as detection device, and using two types of lasers (Ar-ion and Nd-YAG) were carried out, yielding sequencing traces which have quality factor of 20 for read lengths as long as 650 base pairs.

  17. A dual amplification fluorescent strategy for sensitive detection of DNA methyltransferase activity based on strand displacement amplification and DNAzyme amplification.

    PubMed

    Cui, Wanling; Wang, Lei; Jiang, Wei

    2016-03-15

    DNA methyltransferase (MTase) plays a critical role in many biological processes and has been regarded as a predictive cancer biomarker and a therapeutic target in cancer treatment. Sensitive detection of DNA MTase activity is essential for early cancer diagnosis and therapeutics. Here, we developed a dual amplification fluorescent strategy for sensitive detection of DNA MTase activity based on strand displacement amplification (SDA) and DNAzyme amplification. A trifunctional double-stranded DNA (dsDNA) probe was designed including a methylation site for DNA MTase recognition, a complementary sequence of 8-17 DNAzyme for synthesizing DNAzyme, and a nicking site for nicking enzyme cleavage. Firstly, the trifunctional dsDNA probe was methylated by DNA MTase to form the methylated dsDNA. Subsequently, HpaII restriction endonuclease specifically cleaved the residue of unmethylated dsDNA. Next, under the action of polymerase and nicking enzyme, the methylared dsDNA initiated SDA, releasing numbers of 8-17 DNAzymes. Finally, the released 8-17 DNAzymes triggered DNAzyme amplification reaction to induce a significant fluorescence enhancement. This strategy could detect DNA MTase activity as low as 0.0082U/mL. Additionally, the strategy was successfully applied for evaluating the inhibitions of DNA MTase using two anticancer drugs, 5-azacytidine and 5-aza-2'-deoxycytidine. The results indicate the proposed strategy has a potential application in early cancer diagnosis and therapeutics.

  18. High-sensitivity capillary electrophoresis of double-stranded DNA fragments using monomeric and dimeric fluorescent intercalating dyes

    SciTech Connect

    Zhu, H.; Clark, S.M.; Benson, S.C.; Rye, H.S.; Glazer, A.N.; Mathies, R.A. )

    1994-07-01

    Fluorescence-detected capillary electrophoresis separations of [phi]X174/HaeIII DNA restriction fragments have been performed using monomeric and dimeric intercalating dyes. Replaceable hydroxyethyl cellulose solutions were used as the separation medium. Confocal fluorescence detection was performed following 488-nm laser excitation. The limits of DNA detection for on-column staining with monomeric dyes (ethidium bromide, two propidium dye derivatives, oxazole yellow, thiazole orange, and a polycationic thiazole orange derivative) were determined. The thiazole orange dyes provide the most sensitive detection with limiting sensitivities of 2-4 amol of DNA base pairs per band, and detection of the 603-bp fragment was successful, injecting from [phi]X174/HaeIII samples containing only 1-2 fg of this fragment per microliter. Separations of preformed DNA-dimeric dye complexes were also performed. The breadth of the bands observed in separations of preformed DNA-dimeric dye complexes is due to the presence of DNA fragments with different numbers of bound dye molecules that can be resolved as closely spaced subbands in many of our separations. The quality of these DNA-dye complex separations can be dramatically improved by performing the electrophoresis with 9-aminoacridine (9AA) in the column and running buffers. 43 refs., 10 figs., 1 tab.

  19. Using fluorescence immunochromatographic test strips based on quantum dots for the rapid and sensitive determination of microcystin-LR.

    PubMed

    Sun, Jiadi; Li, Ying; Pi, Fuwei; Ji, Jian; Zhang, Yinzhi; Sun, Xiulan

    2017-03-01

    A novel immunosensor for the detection of microcystin-LR (MC-LR) was constructed with use of immunochromatographic test strips (ICTS). Quantum dots were chosen to be the fluorescent labels for the immune sensor in ICTS because of their excellent optical and electronic properties with a relatively narrow emission spectrum. The detection sensitivity of the ICTS was related to the concentration of the fluorescent probe and the amount of the MC-LR standards. Under optimal conditions, with MC-LR as the target, the ICTS sensor had a linear range from 0.25 to 5 μg/L, with a correlation coefficient of 0.9901 and a detection limit of 0.1 μg/L. Furthermore, the repeatability of the ICTS was good, and the coefficient of variation was 10%. The ICTS immunosensor allows the reliable detection of MC-LR in water, and has potential in simple, sensitive detection applications. Graphical Abstract A novel method was developed to detect MC-LR using QDs based immunochromatographic test strip.

  20. Polydopamine Thin Films as Protein Linker Layer for Sensitive Detection of Interleukin-6 by Surface Plasmon Enhanced Fluorescence Spectroscopy.

    PubMed

    Toma, Mana; Tawa, Keiko

    2016-08-31

    Polydopamine (PDA) thin films are introduced to the surface modification of biosensor surfaces utilizing surface plasmon enhanced fluorescence spectroscopy (SPFS) as the linker layer of capture antibody on to the sensor surfaces. The capture antibody can be directly attached to the sensor surface without using any coupling agent by functionalizing the gold sensor surface with PDA thin films. The PDA coating is performed by a single-step preparation process by applying the dopamine solution on the sensor surface, which requires an extremely short incubation time (10 min). The real-time in situ measurement of the adsorption kinetics of the capture antibody onto the PDA-coated sensor surface is studied by surface plasmon resonance (SPR) spectroscopy. It reveals that the immobilization of capture antibody immediately occurs after introduction of a solution containing capture antibody, and the sensor surface is fully covered with the capture antibody. The sensitive detection of the cytokine marker interleukin-6 (IL-6) is performed by SPFS using a sandwich assay format with fluorescently labeled detection antibody. The sensor chips functionalized by PDA chemistry exhibited sensitive sensor responses with low nonspecific adsorption of the detection antibody onto the sensor surface. The detection limit of IL-6 with the developed SPFS biosensor is determined to be 2 pg/mL (100 fM), which is within the range of the diagnostic criteria. Our observation elucidates the remarkable utility of PDA coatings for chemical modification of the metallic sensor surfaces by a simple, brief, and inexpensive manner.

  1. Sets of RNA Repeated Tags and Hybridization-Sensitive Fluorescent Probes for Distinct Images of RNA in a Living Cell

    PubMed Central

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

    2010-01-01

    Background Imaging the behavior of RNA in a living cell is a powerful means for understanding RNA functions and acquiring spatiotemporal information in a single cell. For more distinct RNA imaging in a living cell, a more effective chemical method to fluorescently label RNA is now required. In addition, development of the technology labeling with different colors for different RNA would make it easier to analyze plural RNA strands expressing in a cell. Methodology/Principal Findings Tag technology for RNA imaging in a living cell has been developed based on the unique chemical functions of exciton-controlled hybridization-sensitive oligonucleotide (ECHO) probes. Repetitions of selected 18-nucleotide RNA tags were incorporated into the mRNA 3′-UTR. Pairs with complementary ECHO probes exhibited hybridization-sensitive fluorescence emission for the mRNA expressed in a living cell. The mRNA in a nucleus was detected clearly as fluorescent puncta, and the images of the expression of two mRNAs were obtained independently and simultaneously with two orthogonal tag–probe pairs. Conclusions/Significance A compact and repeated label has been developed for RNA imaging in a living cell, based on the photochemistry of ECHO probes. The pairs of an 18-nt RNA tag and the complementary ECHO probes are highly thermostable, sequence-specifically emissive, and orthogonal to each other. The nucleotide length necessary for one tag sequence is much shorter compared with conventional tag technologies, resulting in easy preparation of the tag sequences with a larger number of repeats for more distinct RNA imaging. PMID:20885944

  2. Highly sensitive rapid fluorescence detection of protein residues on surgical instruments

    NASA Astrophysics Data System (ADS)

    Kovalev, Valeri I.; Bartona, James S.; Richardson, Patricia R.; Jones, Anita C.

    2006-07-01

    There is a risk of contamination of surgical instruments by infectious protein residues, in particular, prions which are the agents for Creutzfeldt-Jakob Disease in humans. They are exceptionally resistant to conventional sterilization, therefore it is important to detect their presence as contaminants so that alternative cleaning procedures can be applied. We describe the development of an optimized detection system for fluorescently labelled protein, suitable for in-hospital use. We show that under optimum conditions the technique can detect ~10 attomole/cm2 with a scan speed of ~3-10 cm2/s of the test instrument's surface. A theoretical analysis and experimental measurements will be discussed.

  3. DNA nicks and increased sensitivity of DNA to fluorescence in situ end labeling during functional spermiogenesis.

    PubMed

    Smith, A; Haaf, T

    1998-09-01

    Terminal transferase can be used to quantitate DNA strand breaks in situ by labeling free 3'-hydroxyl ends with exogenous nucleotides. Endogenous nicks in DNA temporally appear and disappear during functionally significant structural rearrangements of chromatin. Fluorescence in situ end labeling of mouse and rat testicular cells demonstrated that functional spermiogenesis is associated with abundant DNA nicks that occur in elongating spermatids, most likely as a result of nucleoprotein changes during terminal differentiation. Detectable DNA breaks were not observed in round spermatids and epididymal sperm.

  4. Phase-Sensitive Fluorescence Study of Mono-L-Aspartyl Chlorin E6

    DTIC Science & Technology

    2007-11-02

    evolving as a modality for the treatment of malignant tumors. Most of the clinical work on PDT has been carried out, using hematoporphyrin derivative...HpD). [1,2] HpD is effective as a photosensitizer of PDT , but has some problems to be overcome. One of them is that HpD does not absorb...results of phase-resolved fluorescence measurement on ME2906, which is currently an important photosensitizer in PDT . In this study, we have done the

  5. Gadolinium(III)-sensitized fluorescence of europium in its mixed-metal compounds with trifluroacetate

    NASA Astrophysics Data System (ADS)

    Kalinovskaya, I. V.; Zadorozhnaya, A. N.

    2017-04-01

    The fluorescence properties of mixed-metal compounds of Eu(III) and Gd(III) with trifluoroacetic acid, Eu1-xGdx(C2F3O2)3·yD·zH2O, where D - 1,10-phenanthroline, 2,2-dipyridil, diphenylguanidine, x = 0, 0.25, 0.5, or 0.7, were studied. Luminescence spectroscopic evidence and the examination of excitation spectra indicate the occurrence of efficient energy transfer from the gadolinium to the europium ion. The greatest promotion of Eu3+ photoluminescence at 615 nm is observed when Eu:Gd = 1:1.

  6. Use of a beta microprobe system to measure arterial input function in PET via an arteriovenous shunt in rats

    PubMed Central

    2011-01-01

    Background Kinetic modeling of physiological function using imaging techniques requires the accurate measurement of the time-activity curve of the tracer in plasma, known as the arterial input function (IF). The measurement of IF can be achieved through manual blood sampling, the use of small counting systems such as beta microprobes, or by derivation from PET images. Previous studies using beta microprobe systems to continuously measure IF have suffered from high background counts. Methods In the present study, a light-insensitive beta microprobe with a temporal resolution of up to 1 s was used in combination with a pump-driven femoral arteriovenous shunt to measure IF in rats. The shunt apparatus was designed such that the placement of the beta microprobe was highly reproducible. The probe-derived IF was compared to that obtained from manual sampling at 5-s intervals and IF derived from a left ventricle VOI in a dynamic PET image of the heart. Results Probe-derived IFs were very well matched to that obtained by "gold standard" manual blood sampling, but with an increased temporal resolution of up to 1 s. The area under the curve (AUC) ratio between probe- and manually derived IFs was 1.07 ± 0.05 with a coefficient of variation of 0.04. However, image-derived IFs were significantly underestimated compared to the manually sampled IFs, with an AUC ratio of 0.76 ± 0.24 with a coefficient of variation of 0.32. Conclusions IF derived from the beta microprobe accurately represented the IF as measured by blood sampling, was reproducible, and was more accurate than an image-derived technique. The use of the shunt removed problems of tissue-background activity, and the use of a light-tight probe with minimal gamma sensitivity refined the system. The probe/shunt apparatus can be used in both microprobe and PET studies. PMID:22214227

  7. Study of microstructure and silicon segregation in cast iron using color etching and electron microprobe analysis

    SciTech Connect

    Vazehrad, S.; Diószegi, A.

    2015-06-15

    An investigation on silicon segregation of lamellar, compacted and nodular graphite iron was carried out by applying a selective, immersion color etching and a modified electron microprobe to study the microstructure. The color etched micrographs of the investigated cast irons by revealing the austenite phase have provided data about the chronology and mechanism of microstructure formation. Moreover, electron microprobe has provided two dimensional segregation maps of silicon. A good agreement was found between the segregation profile of silicon in the color etched microstructure and the silicon maps achieved by electron microprobe analysis. However, quantitative silicon investigation was found to be more accurate than color etching results to study the size of the eutectic colonies. - Highlights: • Sensitivity of a color etchant to silicon segregation is quantitatively demonstrated. • Si segregation measurement by EMPA approved the results achieved by color etching. • Color etched micrographs provided data about solidification mechanism in cast irons. • Austenite grain boundaries were identified by measuring the local Si concentration.

  8. Fluorescent carbon nanodots for sensitive and selective detection of tannic acid in wines.

    PubMed

    Ahmed, Gaber Hashem Gaber; Laíño, Rosana Badía; Calzón, Josefa Angela García; García, Marta Elena Díaz

    2015-01-01

    Herein we describe an easy one step synthesis of carbon nanodots (C-dots) by thermal carbonization of 6-bromohexylboronic acid using two different amine compounds, polyethyleneglycol bis(3-aminopropyl (PEGA) and 1,2-aminopropane (DPA), at 180 °C in atmospheric oxygen. The as-synthesized C-dots were characterized by FTIR, HRTEM, NMR and fluorescence. The C-dots prepared using PEGA showed a strong emission at 440 nm with excitation at 362 nm. These C-dots exhibited analytical potential as sensing probes for tannic acid (TA) determination. pH effect, interferences, and analytical performance of the method were investigated. The method was found effective in the linear concentration range from 0.1 to 10 mg L(-1) TA achieving a limit of detection equal 0.018 mg L(-1) TA. The applicability of the method was demonstrated by direct measurements of TA in red and white wine samples. Validation of the method was achieved by spiking the wine samples with different standard TA concentrations obtaining recoveries in the range (90-112.5%). A probable mechanism by which TA quenched the C-dots fluorescence was proposed.

  9. Sensitive detection of copper ions via ion-responsive fluorescence quenching of engineered porous silicon nanoparticles

    NASA Astrophysics Data System (ADS)

    Hwang, Jangsun; Hwang, Mintai P.; Choi, Moonhyun; Seo, Youngmin; Jo, Yeonho; Son, Jaewoo; Hong, Jinkee; Choi, Jonghoon

    2016-10-01

    Heavy metal pollution has been a problem since the advent of modern transportation, which despite efforts to curb emissions, continues to play a critical role in environmental pollution. Copper ions (Cu2+), in particular, are one of the more prevalent metals that have widespread detrimental ramifications. From this perspective, a simple and inexpensive method of detecting Cu2+ at the micromolar level would be highly desirable. In this study, we use porous silicon nanoparticles (NPs), obtained via anodic etching of Si wafers, as a basis for undecylenic acid (UDA)- or acrylic acid (AA)-mediated hydrosilylation. The resulting alkyl-terminated porous silicon nanoparticles (APS NPs) have enhanced fluorescence stability and intensity, and importantly, exhibit [Cu2+]-dependent quenching of fluorescence. After determining various aqueous sensing conditions for Cu2+, we demonstrate the use of APS NPs in two separate applications – a standard well-based paper kit and a portable layer-by-layer stick kit. Collectively, we demonstrate the potential of APS NPs in sensors for the effective detection of Cu2+.

  10. Sensitive detection of copper ions via ion-responsive fluorescence quenching of engineered porous silicon nanoparticles

    PubMed Central

    Hwang, Jangsun; Hwang, Mintai P.; Choi, Moonhyun; Seo, Youngmin; Jo, Yeonho; Son, Jaewoo; Hong, Jinkee; Choi, Jonghoon

    2016-01-01

    Heavy metal pollution has been a problem since the advent of modern transportation, which despite efforts to curb emissions, continues to play a critical role in environmental pollution. Copper ions (Cu2+), in particular, are one of the more prevalent metals that have widespread detrimental ramifications. From this perspective, a simple and inexpensive method of detecting Cu2+ at the micromolar level would be highly desirable. In this study, we use porous silicon nanoparticles (NPs), obtained via anodic etching of Si wafers, as a basis for undecylenic acid (UDA)- or acrylic acid (AA)-mediated hydrosilylation. The resulting alkyl-terminated porous silicon nanoparticles (APS NPs) have enhanced fluorescence stability and intensity, and importantly, exhibit [Cu2+]-dependent quenching of fluorescence. After determining various aqueous sensing conditions for Cu2+, we demonstrate the use of APS NPs in two separate applications – a standard well-based paper kit and a portable layer-by-layer stick kit. Collectively, we demonstrate the potential of APS NPs in sensors for the effective detection of Cu2+. PMID:27752120

  11. Fluorescence-guided tumor visualization using a custom designed NIR attachment to a surgical microscope for high sensitivity imaging (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Kittle, David S.; Patil, Chirag G.; Mamelak, Adam; Hansen, Stacey; Perry, Jeff; Ishak, Laura; Black, Keith L.; Butte, Pramod V.

    2016-03-01

    Current surgical microscopes are limited in sensitivity for NIR fluorescence. Recent developments in tumor markers attached with NIR dyes require newer, more sensitive imaging systems with high resolution to guide surgical resection. We report on a small, single camera solution enabling advanced image processing opportunities previously unavailable for ultra-high sensitivity imaging of these agents. The system captures both visible reflectance and NIR fluorescence at 300 fps while displaying full HD resolution video at 60 fps. The camera head has been designed to easily mount onto the Zeiss Pentero microscope head for seamless integration into surgical procedures.

  12. Fluorescent probes for "off-on" highly sensitive detection of Hg²⁺ and L-cysteine based on nitrogen-doped carbon dots.

    PubMed

    Zhang, Yi; Cui, Peipei; Zhang, Feng; Feng, Xiaoting; Wang, Yaling; Yang, Yongzhen; Liu, Xuguang

    2016-05-15

    Fluorescent nitrogen-doped carbon dots (NCDs) were synthesized by a facile, and low-cost one-step hydrothermal strategy using citric acid as carbon source and ammonia solution as nitrogen source for the first time. The obtained NCDs show stable blue fluorescence with a high quantum yield of 35.4%, along with the fluorescence lifetime of ca. 6.75 ns. Most importantly, Hg(2+) can completely quench the fluorescence of NCDs as a result of the formation of a non-fluorescent stable NCDs-Hg(2+) complex. Static fluorescence quenching towards Hg(2+) is proved by the Stern-Volmer equation, ultraviolet-visible absorption spectra, temperature dependent quenching and fluorescence lifetime measurements. Subsequently, the fluorescence of the NCDs-Hg(2+) system is completely recovered with the addition L-cysteine (L-Cys) owing to the dissociation of NCDs-Hg(2+) complex to form a more stable Hg(2+)-L-Cys complex by Hg(2+)-S bonding. Therefore, such NCDs can be used as an effective fluorescent "turn-off" probe for rapid, rather highly selective and sensitive detection of Hg(2+), with a limit of detection (LOD) as low as 1.48 nM and a linear detection range of 0-10 μM. Interestingly, NCDs-Hg(2+) system can be conveniently employed as a fluorescent "turn-on" sensor for highly selective and sensitive detection of L-Cys with a low LOD of 0.79 nM and a wide linear detection range of 0-50 μM. Further, the sensitivity of NCDs to Hg(2+) is preserved in tap water with a LOD of 1.65 nM and a linear detection range of 0-10 μM.

  13. Structure-matched Phthalocyanine Ion Pair as a Red-emitting Fluorescent Optical Probe for the Analysis of Sodium Dodecylbenzenesulfonate with High Specificity and Sensitivity.

    PubMed

    Yu, Fei; Guo, Menglin; Deng, Yabin; Lu, Yin; Chen, Lin; Huang, Ping; Li, Donghui

    2016-01-01

    We have found that a positively charged cationic copper phthalocyanine, Alcian blue (Alcian blue 8GX), can efficiently quench the fluorescence of an oppositely charged red fluorescent phthalocyanine compound with a matched molecular structure, tetrasulfonated aluminum phthalocyanine (AlS4Pc), because of the formation of an ion pair complex (AlS4Pc-Alcian blue 8GX) that exhibits almost no fluorescence. An investigation was carried out on the fluorescence recovery of AlS4Pc-Alcian blue 8GX caused by a series of anionic surfactants containing a sulfonic group (sodium dodecylbenzenesulfonate (SDBS), sodium lauryl sulfate (SLS), and sodium dodecyl sulfate (SDS)). The results showed that SDBS exhibited a significant response, and the highest sensitivity among the surfactants. Due to its high efficiency of fluorescence quenching and the high level of fluorescence recovery, direct observes can even be performed by the naked eye. The results revealed that the Alcian blue 8GX-AlS4Pc ion-pair complex fluorescent probe only responded to SDBS in the low-concentration range. Based on the new founding, this study proposed a novel principle and method of fluorescence enhancement to specifically measure the concentration of SDBS, thereby achieving a highly sensitive and highly specific determination of SDBS. Under the optimal conditions, the fluorescence intensity (I(f)) of the system and the concentration of SDBS in the range of 1 × 10(-7) - 1 × 10(-5) mol/dm(3) exhibited a good linear relationship. This method is highly sensitive, and the operation is simple and rapid. It had been applied for the quantitative analysis of SDBS in environmental water, while achieving satisfactory results compared with those of the standard method. This study developed a new application of the fluorescent phthalocyanine compounds used as molecular probes in analytical sciences.

  14. Hg diffusion in books of XVIII and XIX centuries by synchrotron microprobe

    NASA Astrophysics Data System (ADS)

    Pessanha, S.; Carvalho, M. L.; Manso, M.; Guilherme, A.; Marques, A. F.; Perez, C. A.

    2009-08-01

    The pigment vermilion (HgS) was used to color the fore edge, tail and head of books. Dissemination and quantification of Hg present in the ink used to color books from XVIII and XIX centuries are reported. Mercury is a very toxic element for the human body, therefore it is extremely important to know whether Hg tends to disseminate throughout the paper or stays confined to the borders of the books with less danger for readers. Synchrotron X-ray microprobe was used to evaluate Hg dissemination from the border to the centre of the paper sheet. The diffusion pattern of Hg was compared with the results obtained by a portable X-ray fluorescence spectrometer and mean quantitative calculations were obtained by a stationary X-ray fluorescence system with triaxial geometry. The results showed high concentrations of Hg in the external regions, but no diffusion was observed for the inner parts of the paper.

  15. A DNA machine-based fluorescence amplification strategy for sensitive detection of uracil-DNA glycosylase activity.

    PubMed

    Wu, Yushu; Wang, Lei; Zhu, Jing; Jiang, Wei

    2015-06-15

    Sensitive detection of uracil-DNA glycosylase (UDG) activity is critical for function study of UDG and clinical diagnosis. Here, we developed a novel fluorescent strategy for sensitive detection of UDG activity based on the signal amplification by a label-free and enzyme-free DNA machine. A double-strand DNA (dsDNA) probe P1-P2 with uracil bases and trigger sequence was designed for UDG recognition and signal transduction. Two hairpin probes H1 and H2 which were partially complementary were employed to construct the label-free and enzyme-free DNA machine. Under the action of UDG, uracil bases were removed from the P1-P2 dsDNA probe, and then a strand P2' with abasic sites was released. Subsequently, the liberated P2' activated the DNA machine and generated numerous H1-H2 complexes containing G-quadruplex (G4) structures in the end. Finally, the G4 structures could bind with N-methylmesoporphyrin IX (NMM) to form G4-NMM complexes with the enhanced fluorescence responses. This strategy could detect UDG activity as low as 0.00044 U/mL. In addition, the strategy was also applied for the analysis of UDG activity in HeLa cells lysate with low effect of cellular components. Moreover, this strategy was successfully applied for assaying the inhibition of UDG using uracil glycosylase inhibitor (UGI). This strategy provided a potential tool for sensitive quantification of UDG activity in UDG functional study and clinical diagnosis.

  16. Toehold-mediated strand displacement reaction-dependent fluorescent strategy for sensitive detection of uracil-DNA glycosylase activity.

    PubMed

    Wu, Yushu; Wang, Lei; Jiang, Wei

    2017-03-15

    Sensitive detection of uracil-DNA glycosylase (UDG) activity is beneficial for evaluating the repairing process of DNA lesions. Here, toehold-mediated strand displacement reaction (TSDR)-dependent fluorescent strategy was constructed for sensitive detection of UDG activity. A single-stranded DNA (ssDNA) probe with two uracil bases and a trigger sequence were designed. A hairpin probe with toehold domain was designed, and a reporter probe was also designed. Under the action of UDG, two uracil bases were removed from ssDNA probe, generating apurinic/apyrimidinic (AP) sites. Then, the AP sites could inhibit the TSDR between ssDNA probe and hairpin probe, leaving the trigger sequence in ssDNA probe still free. Subsequently, the trigger sequence was annealed with the reporter probe, initiating the polymerization and nicking amplification reaction. As a result, numerous G-quadruplex (G4) structures were formed, which could bind with N-methyl-mesoporphyrin IX (NMM) to generate enhanced fluorescent signal. In the absence of UDG, the ssDNA probe could hybridize with the toehold domain of the hairpin probe to initiate TSDR, blocking the trigger sequence, and then the subsequent amplification reaction would not occur. The proposed strategy was successfully implemented for detecting UDG activity with a detection limit of 2.7×10(-5)U/mL. Moreover, the strategy could distinguish UDG well from other interference enzymes. Furthermore, the strategy was also applied for detecting UDG activity in HeLa cells lysate with low effect of cellular components. These results indicated that the proposed strategy offered a promising tool for sensitive quantification of UDG activity in UDG-related function study and disease prognosis.

  17. A cobalt oxyhydroxide-modified upconversion nanosystem for sensitive fluorescence sensing of ascorbic acid in human plasma

    NASA Astrophysics Data System (ADS)

    Cen, Yao; Tang, Jun; Kong, Xiang-Juan; Wu, Shuang; Yuan, Jing; Yu, Ru-Qin; Chu, Xia

    2015-08-01

    Ascorbic acid (AA), a potent antioxidant readily scavenging reactive species, is a crucial micronutrient involved in many biochemical processes. Here, we have developed a cobalt oxyhydroxide (CoOOH)-modified upconversion nanosystem for fluorescence sensing of AA activity in human plasma. The nanosystem consists of upconversion nanoparticles (UCNPs) NaYF4:30% Yb,0.5% Tm@NaYF4, which serve as energy donors, and CoOOH nanoflakes formed on the surface of UCNPs, which act as efficient energy acceptors. The fluorescence resonance energy transfer (FRET) process from the UCNPs to the absorbance of the CoOOH nanoflakes occurs in the nanosystem. The AA-mediated specific redox reaction reduces CoOOH into Co2+, leading to the inhibition of FRET, and resulting in the recovery of upconversion emission spectra. On the basis of these features, the nanosystem can be used for sensing AA activity with sensitivity and selectivity. Moreover, due to the minimizing background interference provided by UCNPs, the nanosystem has been applied to monitoring AA levels in human plasma sample with satisfactory results. The proposed approach may potentially provide an analytical platform for research and clinical diagnosis of AA related diseases.Ascorbic acid (AA), a potent antioxidant readily scavenging reactive species, is a crucial micronutrient involved in many biochemical processes. Here, we have developed a cobalt oxyhydroxide (CoOOH)-modified upconversion nanosystem for fluorescence sensing of AA activity in human plasma. The nanosystem consists of upconversion nanoparticles (UCNPs) NaYF4:30% Yb,0.5% Tm@NaYF4, which serve as energy donors, and CoOOH nanoflakes formed on the surface of UCNPs, which act as efficient energy acceptors. The fluorescence resonance energy transfer (FRET) process from the UCNPs to the absorbance of the CoOOH nanoflakes occurs in the nanosystem. The AA-mediated specific redox reaction reduces CoOOH into Co2+, leading to the inhibition of FRET, and resulting in the

  18. Highly sensitive fluorescent stain for detecting lipopolysaccharides in sodium dodecyl sulfate polyacrylamide gel electrophoresis.

    PubMed

    Wang, Xu; Zhou, Ayi; Cai, Wanhui; Yu, Dongdong; Zhu, Zhongxin; Jiang, Chengxi; Jin, Litai

    2015-08-01

    A sensitive and simple technique was developed for the visualization of gel-separated lipopolysaccharides by using a hydrazide derivative, UGF202. As low as 0.5-1 ng total LPS could be detected by UGF202 stain, which is 2- and 16-fold more sensitive than that of the commonly used Pro-Q Emerald 300 and Keenan et al. developed silver stain, respectively. The results indicated that UGF202 stain could be a good choice for LPS determination in polyacrylamide gels.

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

    PubMed

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

    2016-02-01

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

  20. Highly sensitive rapid fluorescence detection of protein residues on surgical instruments

    NASA Astrophysics Data System (ADS)

    Kovalev, Valeri I.; Barton, James S.; Richardson, Patricia R.; Jones, Anita C.

    2006-02-01

    There is a risk of contamination of surgical instruments by nfectious protein residues, in particular, prions which are the agents for Creutzfeldt-Jakob Disease in humans. They are exceptionally resistant to conventional sterilization, therefore it is important to detect their presence as contaminants so that alternative cleaning procedures can be applied. We describe the development of an optimized detection system for fluorescently labelled protein, suitable for in-hospital use. We show that under optimum conditions the technique can detect ~100 zeptomoles/mm2 with an area scan speed of ~20 cm2/s and for using the system to detect other agents of biomedical interest. A theoretical analysis and experimental measurements will be discussed.

  1. Utilizing a pH-Sensitive Dye in the Selective Fluorescent Recognition of Sulfate.

    PubMed

    Agafontsev, Aleksandr M; Shumilova, Tatiana A; Panchenko, Pavel A; Janz, Sharon; Fedorova, Olga A; Kataev, Evgeny A

    2016-10-10

    A receptor containing amidopyrrole binding subunits and free amino groups, conjugated to a naphthalimide dye, has been designed and synthesized. The intrinsic selectivity of the binding motif for phosphate present in DMSO completely disappears in 10 % DMSO aqueous buffer at pH 3.6, at which the receptor is protonated. The electrostatic interactions between the receptor and an anion start to dominate, thus leading to selectivity for sulfate. The ability of the HSO4(-) anion to transfer the proton to the amino group during the recognition event suppresses the photoinduced electron transfer (PET) on the dye, resulting in a selective turn-on fluorescent response. The choice of pH of the solution for sensing is dictated by the pKa value of the dye.

  2. A micropatterned cell array with an integrated oxygen-sensitive fluorescent membrane.

    PubMed

    Montagne, Kevin; Komori, Kikuo; Yang, Fei; Tatsuma, Tetsu; Fujii, Teruo; Sakai, Yasuyuki

    2009-11-01

    We propose a simple method for producing micropatterned cell spots by photocatalytic lithography on a Pt porphyrin-based oxygen-sensitive polystyrene membrane that enables real-time imaging of oxygen consumption of patterned cell spots with sub-millimetre resolution.

  3. Fluorescence polarization immunoassays for rapid, accurate, and sensitive determination of mycotoxins

    Technology Transfer Automated Retrieval System (TEKTRAN)

    Analytical methods for the determination of mycotoxins in foods are commonly based on chromatographic techniques (GC, HPLC or LC-MS). Although these methods permit a sensitive and accurate determination of the analyte, they require skilled personnel and are time-consuming, expensive, and unsuitable ...

  4. Sensitive fluorescent detection of DNA methyltransferase using nicking endonuclease-mediated multiple primers-like rolling circle amplification.

    PubMed

    Huang, Juan; Li, Xiao-Yu; Du, Yi-Chen; Zhang, Li-Na; Liu, Ke-Ke; Zhu, Li-Na; Kong, De-Ming

    2017-05-15

    Sensitive and reliable detection of DNA methyltransferase (MTase) is of great significance for both early tumor diagnosis and therapy. In this study, a simple, label-free and sensitive DNA MTase-sensing method was developed on the basis of a nicking endonuclease-mediated multiple primers-like rolling circle amplification (RCA) strategy. In this method, a dumbbell RCA template was prepared by blunt-end ligation of two molecules of hairpin DNA. In addition to the primer-binding sequence, the dumbbell template contained another three important parts: 5'-CCGG-3' sequences in double-stranded stems, nicking endonuclease recognition sites and C-rich sequences in single-stranded loops. The introduction of 5'-CCGG-3' sequences allows the dumbbell template to be destroyed by the restriction endonuclease, HpaII, but is not destroyed in the presence of the target MTase-M.SssI MTase. The introduction of nicking endonuclease recognition sites makes the M.SssI MTase-protected dumbbell template-mediated RCA proceed in a multiple primers-like exponential mode, thus providing the RCA with high amplification efficiency. The introduction of C-rich sequences may promote the folding of amplification products into a G-quadruplex structure, which is specifically recognized by the commercially available fluorescent probe thioflavin T. Improved RCA amplification efficiency and specific fluorescent recognition of RCA products provide the M.SssI MTase-sensing platform with high sensitivity. When a dumbbell template containing four nicking endonuclease sites is used, highly specific M.SssI MTase activity detection can be achieved in the range of 0.008-50U/mL with a detection limit as low as 0.0011U/mL. Simple experimental operation and mix-and-detection fluorescent sensing mode ensures that M.SssI MTase quantitation works well in a real-time RCA mode, thus further simplifying the sensing performance and making high throughput detection possible. The proposed MTase-sensing strategy was also

  5. Fluorescent-dye-doped sol-gel sensor for highly sensitive carbon dioxide gas detection below atmospheric concentrations.

    PubMed

    Dansby-Sparks, Royce N; Jin, Jun; Mechery, Shelly J; Sampathkumaran, Uma; Owen, Thomas William; Yu, Bi Dan; Goswami, Kisholoy; Hong, Kunlun; Grant, Joseph; Xue, Zi-Ling

    2010-01-15

    Optical fluorescence sol-gel sensors have been developed for the detection of carbon dioxide gas in the 0.03-30% range with a detection limit of 0.008% (or 80 ppm) and a quantitation limit of 0.02% (or 200 ppm) CO(2). Sol-gels were spin-coated on glass slides to create an organically modified silica-doped matrix with the 1-hydroxypyrene-3,6,8-trisulfonate (HPTS) fluorescent indicator. The luminescence intensity of the HPTS indicator (513 nm) is quenched by CO(2), which protonates the anionic form of HPTS. An ion pair technique was used to incorporate the lipophilic dye into the hydrophilic sol-gel matrix. TiO(2) particles (<5 microm diameter) were added to induce Mie scattering and increase the incident light interaction with the sensing film, thus increasing the signal-to-noise ratio. Moisture-proof overcoatings have been used to maintain a constant level of water inside the sensor films. The optical sensors are inexpensive to prepare and can be easily coupled to fiber optics for remote sensing capabilities. A fiber-optic bundle was used for the gas detection and shown to work as part of a multianalyte platform for simultaneous detection of multiple analytes. The studies reported here resulted in the development of sol-gel optical fluorescent sensors for CO(2) gas with sensitivity below that in the atmosphere (ca. 387 ppm). These sensors are a complementary approach to current FT-IR measurements for real-time carbon dioxide detection in environmental applications.

  6. Copper nanoclusters as a highly sensitive and selective fluorescence sensor for ferric ions in serum and living cells by imaging.

    PubMed

    Cao, Haiyan; Chen, Zhaohui; Zheng, Huzhi; Huang, Yuming

    2014-12-15

    A simple, one-step facile route for preparation of water soluble and fluorescent Cu nanoclusters (NCs) stabilized by tannic acid (TA) is described. The as-prepared TA capped Cu NCs (TA-Cu NCs) are characterized by UV-vis spectroscopy, Fourier transform infrared (FT-IR) spectroscopy, luminescence, transmission electron microscopy (TEM), and X-ray photoelectron spectroscopy (XPS). The TA-Cu NCs show luminescence properties having excitation and emission maxima at 360 nm and 430 nm, respectively, with a quantum yield of about 14%. The TA-Cu NCs are very stable even in 0.3 M NaCl, and their luminescent properties show pH independent. The fluorescence (FL) of Cu NCs is strongly quenched by Fe(3+) through an electron transfer mechanism, but not by other metal ions. Furthermore, the FL of the TA-Cu NCs shows no changes with the addition of Fe(2+) or H2O2 individually. On this basis, a facile chemosensor was developed for rapid, reliable, sensitive, and selective sensing of Fe(3+) ions with detection limit as low as 10 nM and a dynamic range from 10 nM to 10 μM. The proposed sensor was successfully used for the determination of iron contents in serum samples. Importantly, the Cu NCs-based FL probe showed long-term stability, good biocompatibility and very low cytotoxicity. It was successfully used for imaging ferric ions in living cells, suggesting the potential application of Cu NCs fluorescent probe in clinical analysis and cell imaging.

  7. Highly sensitive and adaptable fluorescence-quenched pair discloses the substrate specificity profiles in diverse protease families.

    PubMed

    Poreba, Marcin; Szalek, Aleksandra; Rut, Wioletta; Kasperkiewicz, Paulina; Rutkowska-Wlodarczyk, Izabela; Snipas, Scott J; Itoh, Yoshifumi; Turk, Dusan; Turk, Boris; Overall, Christopher M; Kaczmarek, Leszek; Salvesen, Guy S; Drag, Marcin

    2017-02-23

    Internally quenched fluorescent (IQF) peptide substrates originating from FRET (Förster Resonance Energy Transfer) are powerful tool for examining the activity and specificity of proteases, and a variety of donor/acceptor pairs are extensively used to design individual substrates and combinatorial libraries. We developed a highly sensitive and adaptable donor/acceptor pair that can be used to investigate the substrate specificity of cysteine proteases, serine proteases and metalloproteinases. This novel pair comprises 7-amino-4-carbamoylmethylcoumarin (ACC) as the fluorophore and 2,4-dinitrophenyl-lysine (Lys(DNP)) as the quencher. Using caspase-3, caspase-7, caspase-8, neutrophil elastase, legumain, and two matrix metalloproteinases (MMP2 and MMP9), we demonstrated that substrates containing ACC/Lys(DNP) exhibit 7 to 10 times higher sensitivity than conventional 7-methoxy-coumarin-4-yl acetic acid (MCA)/Lys(DNP) substrates; thus, substantially lower amounts of substrate and enzyme can be used for each assay. We therefore propose that the ACC/Lys(DNP) pair can be considered a novel and sensitive scaffold for designing substrates for any group of endopeptidases. We further demonstrate that IQF substrates containing unnatural amino acids can be used to investigate protease activities/specificities for peptides containing post-translationally modified amino acids. Finally, we used IQF substrates to re-investigate the P1-Asp characteristic of caspases, thus demonstrating that some human caspases can also hydrolyze substrates after glutamic acid.

  8. Highly sensitive and adaptable fluorescence-quenched pair discloses the substrate specificity profiles in diverse protease families

    PubMed Central

    Poreba, Marcin; Szalek, Aleksandra; Rut, Wioletta; Kasperkiewicz, Paulina; Rutkowska-Wlodarczyk, Izabela; Snipas, Scott J.; Itoh, Yoshifumi; Turk, Dusan; Turk, Boris; Overall, Christopher M.; Kaczmarek, Leszek; Salvesen, Guy S.; Drag, Marcin

    2017-01-01

    Internally quenched fluorescent (IQF) peptide substrates originating from FRET (Förster Resonance Energy Transfer) are powerful tool for examining the activity and specificity of proteases, and a variety of donor/acceptor pairs are extensively used to design individual substrates and combinatorial libraries. We developed a highly sensitive and adaptable donor/acceptor pair that can be used to investigate the substrate specificity of cysteine proteases, serine proteases and metalloproteinases. This novel pair comprises 7-amino-4-carbamoylmethylcoumarin (ACC) as the fluorophore and 2,4-dinitrophenyl-lysine (Lys(DNP)) as the quencher. Using caspase-3, caspase-7, caspase-8, neutrophil elastase, legumain, and two matrix metalloproteinases (MMP2 and MMP9), we demonstrated that substrates containing ACC/Lys(DNP) exhibit 7 to 10 times higher sensitivity than conventional 7-methoxy-coumarin-4-yl acetic acid (MCA)/Lys(DNP) substrates; thus, substantially lower amounts of substrate and enzyme can be used for each assay. We therefore propose that the ACC/Lys(DNP) pair can be considered a novel and sensitive scaffold for designing substrates for any group of endopeptidases. We further demonstrate that IQF substrates containing unnatural amino acids can be used to investigate protease activities/specificities for peptides containing post-translationally modified amino acids. Finally, we used IQF substrates to re-investigate the P1-Asp characteristic of caspases, thus demonstrating that some human caspases can also hydrolyze substrates after glutamic acid. PMID:28230157

  9. A Novel Sensor for Sensitive and Selective Detection of Iodide Using Turn-on Fluorescence Graphene Quantum Dots/Ag Nanocomposite.

    PubMed

    Xu, Xianghong; Wang, Yanhui

    2015-01-01

    Based on the principle of fluorescence enhancing, by the strong and specific interreaction between iodide (I(-)) ions and nanoAg on the surface of graphene quantum dots/Ag (GQDs/Ag) nanocomposite, we propose a simple label-free and turn-on method for the detection of I(-) ions with high selectivity and sensitivity by using fluorescent GQDs/Ag nanocomposite in aqueous media.

  10. Sensitive detection of sodium cromoglycate with glutathione-capped CdTe quantum dots as a novel fluorescence probe.

    PubMed

    Hao, Chenxia; Liu, Shaopu; Li, Dan; Yang, Jidong; He, Youqiu

    2015-11-01

    A sensitive and simple analytical strategy for the detection of sodium cromoglycate (SCG) has been established based on a readily detectable fluorescence quenching effect of SCG for glutathione-capped (GSH-capped) CdTe quantum dots (QDs). The fluorescence of GSH-capped CdTe QDs could be efficiently quenched by SCG through electron transfer from GSH-capped CdTe QDs to SCG. Under optimum conditions, the response was linearly proportional to the concentration of SCG between 0.6419 and 100 µg/mL, with a correlation coefficient (R) of 0.9964; the detection limit (3δ/K) was 0.1926 µg/mL. The optimum conditions and the influence of coexisting foreign substances on the reaction were also investigated. The very effective and simple method reported here has been successfully applied to the determination of SCG in synthetic and real samples. It is believed that the established approach could have good prospects for application in the fields of clinical diseases diagnosis and treatment.

  11. Cleavable DNA-protein hybrid molecular beacon: A novel efficient signal translator for sensitive fluorescence anisotropy bioassay.

    PubMed

    Hu, Pan; Yang, Bin

    2016-01-15

    Due to its unique features such as high sensitivity, homogeneous format, and independence on fluorescent intensity, fluorescence anisotropy (FA) assay has become a hotspot of study in oligonucleotide-based bioassays. However, until now most FA probes require carefully customized structure designs, and thus are neither generalizable for different sensing systems nor effective to obtain sufficient signal response. To address this issue, a cleavable DNA-protein hybrid molecular beacon was successfully engineered for signal amplified FA bioassay, via combining the unique stable structure of molecular beacon and the large molecular mass of streptavidin. Compared with single DNA strand probe or conventional molecular beacon, the DNA-protein hybrid molecular beacon exhibited a much higher FA value, which was potential to obtain high signal-background ratio in sensing process. As proof-of-principle, this novel DNA-protein hybrid molecular beacon was further applied for FA bioassay using DNAzyme-Pb(2+) as a model sensing system. This FA assay approach could selectively detect as low as 0.5nM Pb(2+) in buffer solution, and also be successful for real samples analysis with good recovery values. Compatible with most of oligonucleotide probes' designs and enzyme-based signal amplification strategies, the molecular beacon can serve as a novel signal translator to expand the application prospect of FA technology in various bioassays.

  12. Highly sensitive fluorescence and SERS detection of azide through a simple click reaction of 8-chloroquinoline and phenylacetylene.

    PubMed

    Zeng, Qing; Ye, Lingling; Ma, Lu; Yin, Wenqing; Li, Tingsheng; Liang, Aihui; Jiang, Zhiliang

    2015-05-01

    In 0.19 mol/L acetic acid (HAc), a click reaction of 8-chloroquinoline/azide/phenylacetylene take places in aqueous solution without Cu(I) as a catalyst. 8-Chloroquinoline (CQN) exhibited a strong fluorescence peak at 430 nm that was quenched linearly as the concentration of azide increased from 20 to 1000 ng/mL. This quenching was due to consumption of CQN in the click reaction and a decrease in the number of efficiently excited photons due to the presence of triazole-quinoline ramification molecules with strong hydrophobicity. Using blue nanosilver sol as the substrate, CQN absorbed onto the surface of nanosilver particles, showing a strong surface-enhanced Raman scattering (SERS) peak at 1585 cm(-1) that decreased linearly as the azide concentration increased from 8 to 500 ng/mL; the detection limit was 4 ng/mL. Thus, two new, simple and sensitive fluorescence and SERS methods have been developed for the determination of azide via the click reaction.

  13. A highly selective and sensitive near-infrared fluorescent probe for imaging of hydrogen sulphide in living cells and mice

    PubMed Central

    Zhang, Ling; Zheng, Xi Emily; Zou, Fang; Shang, Yanguo; Meng, Wenqi; Lai, En; Xu, Zhichen; Liu, Yi; Zhao, Jing

    2016-01-01

    Hydrogen sulphide (H2S), the third endogenous gaseous signalling molecule, has attracted attention in biochemical research. The selective detection of H2S in living systems is essential for studying its functions. Fluorescence detection methods have become useful tools to explore the physiological roles of H2S because of their real-time and non-destructive characteristics. Herein we report a near-infrared fluorescent probe, NIR-HS, capable of tracking H2S in living organisms. With high sensitivity, good selectivity and low cytotoxicity, NIR-HS was able to recognize both the exogenous and endogenous H2S in living cells. More importantly, it realized the visualization of endogenous H2S generated in cells overexpressing cystathionine β-synthase (CBS), one of the enzymes responsible for producing endogenous H2S. The probe was also successfully applied to detect both the exogenous and endogenous H2S in living mice. The superior sensing properties of the probe render it a valuable research tool in the H2S-related medical research. PMID:26743682

  14. Fluorescence detection of Fe(3+) ions in aqueous solution and living cells based on a high selectivity and sensitivity chemosensor.

    PubMed

    Jia, Hongmin; Gao, Xue; Shi, Yu; Sayyadi, Nima; Zhang, Zhiqiang; Zhao, Qi; Meng, Qingtao; Zhang, Run

    2015-01-01

    Although ferric ion (Fe(3+)) performs critical roles in diverse biochemical processes in living systems, its physiological and pathophysiological functions have not been fully explored due to the lack of methods for quantification of Fe(3+) ions in biological system. In this work, a highly sensitive and selective fluorescence chemosensor, L, was developed for the detection of Fe(3+) ions in aqueous solution and in living cells. L was facile synthesized by one step reaction and well characterized by NMR, API-ES, FT-IR, and elementary analysis. The prepared chemosensor displayed excellent selectivity for Fe(3+) ions detection over a wide range of tested metal ions. In the present of Fe(3+) ions, the strong green fluorescence of L was substantially quenched. The 1:1 stoichiometry of the complexation was confirmed by a Job's plot. The association constant (Ka) of L with Fe(3+) was evaluated using the Benesi-Hildebrand method and was found to be 1.36×10(4) M(-1). The MTT assay determined that L exhibits low cytotoxicity toward living cells. Confocal imaging and flow cytometry studies showed that L is readily interiorized by MDA-MB-231 cells through an energy-dependent pathway and could be used to detect of Fe(3+) ions in living cells.

  15. A simple and sensitive surface molecularly imprinted polymers based fluorescence sensor for detection of λ-Cyhalothrin.

    PubMed

    Liu, Chunbo; Song, Zhilong; Pan, Jianming; Yan, Yongsheng; Cao, Zhijing; Wei, Xiao; Gao, Lin; Wang, Juan; Dai, Jiangdong; Meng, Minjia; Yu, Ping

    2014-07-01

    In this study, surface molecularly imprinted YVO4:Eu(3+) nanoparticles with molecular recognitive optosensing activity were successfully prepared by precipitation polymerization using λ-Cyhalothrin (LC) as template molecules, methacrylic acid and ethylene glycol dimethacrylate as the polymerization precursors which could complex with template molecules, and the material has been characterized by SEM, TEM, FT-IR, XRD, TGA and so on. Meanwhile, the as-prepared core-shell structured nanocomposite (YVO4:Eu(3+)@MIPs), which was composed of lanthanide doped YVO4:Eu(3+) as fluorescent signal and surface molecular imprinted polymers as molecular selective recognition sites, could selectively and sensitively optosense the template molecules. After the experimental conditions were optimized, two linear relationship were obtained covering the concentration range of 2.0-10.0 μM and 10.0-90.0 μM, and the limit of detection (LOD) for LC was found to be 1.76 μM. Furthermore, a possible mechanism was put forward to explain the fluorescence quenching of YVO4:Eu(3+)@MIPs. More importantly, the obtained sensor was proven to be suitable for the detection of residues of LC in real examples. And the excellent performance of this sensor will facilitate future development of rapid and high-efficiency detection of LC.

  16. Tuning the Aggregation/Disaggregation Behavior of Graphene Quantum Dots by Structure-Switching Aptamer for High-Sensitivity Fluorescent Ochratoxin A Sensor.

    PubMed

    Wang, Song; Zhang, Yajun; Pang, Guangsheng; Zhang, Yingwei; Guo, Shaojun

    2017-02-07

    The design of graphene quantum dots (GQDs)-aptamer bioconjugates as the new sensing platform is very important for developing high-sensitivity fluorescent biosensors; however, achieving new bioconjugates is still a great challenge. Herein, we report the development of a new high-sensitivity fluorescent aptasensor for the detection of ochratoxin A (OTA) based on tuning aggregation/disaggregation behavior of GQDs by structure-switching aptamers. The fluorescence sensing process for OTA detection involved two key steps: (1) cDNA-aptamer (cDNA, complementary to part of the OTA aptamer) hybridization induced the aggregation of GQD (fluorescence quenching) after cDNA was added into the GQDs-aptamer bioconjugate solution, and (2) the target of OTA triggered disaggregation of GQD aggregates (fluorescence recovery). Such new fluorescent sensing platform can be used to monitor OTA with a linear range of 0 to 1 ng/mL and very low detection limit of 13 pg/mL, which is among the best in all the developed fluorescent nanoparticles-based sensors. Such sensing strategy is also successful in analyzing OTA in practical red wine sample with 94.4-102.7% of recoveries and relative standard deviation in the range of 2.9-5.8%. The present works open a new way for signaling the target-aptamer binding event by tuning aggregation/disaggregation behavior of GQDs-bioconjugates.

  17. Imaging of conformational changes of proteins with a new environment-sensitive fluorescent probe designed for site-specific labeling of recombinant proteins in live cells.

    PubMed

    Nakanishi, J; Nakajima, T; Sato, M; Ozawa, T; Tohda, K; Umezawa, Y

    2001-07-01

    We demonstrate herein a new method for imaging conformational changes of proteins in live cells using a new synthetic environment-sensitive fluorescent probe, 9-amino-6,8-bis(1,3,2-dithioarsolan-2-yl)-5H-benzo[a]phenoxazin-5-one. This fluorescent probe can be attached to recombinant proteins containing four cysteine residues at the i, i + 1, i + 4, and i + 5 positions of an alpha-helix. The specific binding of the fluorescent probe to this 4Cys motif enables fluorescent labeling inside cells by its extracellular administration. The high sensitivity of the fluorophore to its environment enables monitoring of the conformational changes of the proteins in live cells as changes in its fluorescence intensity. The present method was applied to calmodulin (CaM), a Ca2+-binding protein that was well-known to expose hydrophobic domains, depending on the Ca2+ concentration. A recombinant CaM fused at its C-terminal with a helical peptide containing a 4Cys motif was labeled with the fluorescent probe inside live cells. The fluorescence intensity changed reversibly depending on the intracellular Ca2+ concentration, which reflected the conformational change of the recombinant CaM in the live cells.

  18. Liquid chromatography coupled to molecular fluorescence with postcolumn UV sensitization for thimerosal and derivative compounds monitoring in environmental samples.

    PubMed

    Acosta, Gimena; Torres, Sabier; Kaplan, Marcos; Fernández, Liliana P; Pacheco, Pablo H; Gil, Raúl A

    2016-10-01

    A HPLC coupled with molecular fluorescence (MF) spectrometry method for determination of thimerosal (THM, sodium ethylmercurythiosalicylate, C9 H9 HgNaO2 S), and derivatives is proposed. A sensitization of MF was provoked by UV irradiation of analytes in a home-made photoreactor that served as interface between the LC column and MF spectrometer. This method is applied to determination of THM, ethyl mercury, and thiosalicylic acid in samples of pharmaceutical industry effluents, and waters of La Carolina and Jáchal rivers situated in the center-west side of San Luis city and in the east of San Juan city (Middle West, Argentine) where the effluents are dumped. The LODs calculated on basis of 3σ criterion were 1.8, 5, and 0.05 μmol/L for THM, ethyl mercury, and for thiosalicylic acid, respectively.

  19. Highly sensitive and selective detection of Al(III) ions in aqueous buffered solution with fluorescent peptide-based sensor.

    PubMed

    In, Byunggyu; Hwang, Gi Won; Lee, Keun-Hyeung

    2016-09-15

    A fluorescent sensor based on a tripeptide (SerGluGlu) with a dansyl fluorophore detected selectively Al(III) among 16 metal ions in aqueous buffered solutions without any organic cosolvent. The peptide-based sensor showed a highly sensitive turn on response to aluminium ion with high binding affinity (1.84×10(4)M(-1)) in aqueous buffered solutions. The detection limit (230nM, 5.98ppb) of the peptide-based sensor was much lower than the maximum allowable level (7.41μM) of aluminium ions in drinking water demanded by EPA. The binding mode of the peptide sensor with aluminium ions was characterized using ESI mass spectrometry, NMR titration, and pH titration experiments.

  20. Single-molecule-sensitive fluorescence resonance energy transfer in freely-diffusing attoliter droplets

    SciTech Connect

    Rahmanseresht, Sheema; Ramos, Kieran P.; Gamari, Ben D.; Goldner, Lori S.; Milas, Peker

    2015-05-11

    Fluorescence resonance energy transfer (FRET) from individual, dye-labeled RNA molecules confined in freely-diffusing attoliter-volume aqueous droplets is carefully compared to FRET from unconfined RNA in solution. The use of freely-diffusing droplets is a remarkably simple and high-throughput technique that facilitates a substantial increase in signal-to-noise for single-molecular-pair FRET measurements. We show that there can be dramatic differences between FRET in solution and in droplets, which we attribute primarily to an altered pH in the confining environment. We also demonstrate that a sufficient concentration of a non-ionic surfactant mitigates this effect and restores FRET to its neutral-pH solution value. At low surfactant levels, even accounting for pH, we observe differences between the distribution of FRET values in solution and in droplets which remain unexplained. Our results will facilitate the use of nanoemulsion droplets as attoliter volume reactors for use in biophysical and biochemical assays, and also in applications such as protein crystallization or nanoparticle synthesis, where careful attention to the pH of the confined phase is required.

  1. Rewritable Painting Realized from Ambient-Sensitive Fluorescence of ZnO Nanoparticles

    PubMed Central

    Liu, Kai-Kai; Shan, Chong-Xin; He, Gao-Hang; Wang, Ruo-Qiu; Dong, Lin; Shen, De-Zhen

    2017-01-01

    Paper, as one of the most important information carriers, has contributed to the development and transmission of human civilization greatly. Meanwhile, a serious problem of environmental sustainable development caused by the production and utilization of paper has been resulted to modern society. Therefore, a simple and green route is urgently demanded to realize rewritable painting on paper. Herein, a simple route to rewritable painting on copy paper has been demonstrated by using eco-friendly ZnO nanoparticles (NPs) as fluorescent ink, and vinegar and soda that are frequently used in kitchen as erasing and neutralizing agents. Words or patterns written using the ZnO NPs as ink can be erased by vinegar vapour within five seconds, and after a neutralizing process in the ambient of soda vapour, the paper can be used for writing again. It is worth noting that the resolution and precision of the patterns produced via the above route degrade little after ten rewriting cycles, and the quality of the patterns produced using the ZnO NPs as ink fades little after being storage for several months, which promises the versatile potential applications of the rewriting route proposed in this paper. PMID:28169344

  2. Microwave-accelerated metal-enhanced fluorescence: an ultra-fast and sensitive DNA sensing platform.

    PubMed

    Aslan, Kadir; Malyn, Stuart N; Bector, Geetika; Geddes, Chris D

    2007-11-01

    In this paper, we investigated the effects of low-power microwave heating on the components of the recently described new approach to surface DNA hybridization assays, based on the Microwave-Accelerated Metal-Enhanced Fluorescence (MAMEF) platform technology. Thiolated oligonucleotides have been linked to surface-bound silver nanostructures which partially coat a glass slide. The addition of a complementary fluorescein-labeled oligonucleotide results in metal-enhanced fluorescein emission as the probe is brought into close proximity to the silver upon hybridization. In addition, the combined use with low-power microwave heating, which is thought to locally heat around the silvered surface, affords for both the assay kinetics and optical amplification to also be localized to the surface. In our model DNA target assay reported here, we can detect 23-mer targets in less than 20 s, up to a 600-fold decrease in the assay run time as compared to control samples hybridized to completion at room temperature. Importantly, the use of MAMEF also reduces the extent of unwanted non-specific DNA absorption, further increasing specific DNA target detection limits. It was also found that low-power microwave heating did not denature DNA and the bulk temperature increase near to silver nanoparticles was only ca. 1 degrees C.

  3. Rewritable Painting Realized from Ambient-Sensitive Fluorescence of ZnO Nanoparticles

    NASA Astrophysics Data System (ADS)

    Liu, Kai-Kai; Shan, Chong-Xin; He, Gao-Hang; Wang, Ruo-Qiu; Dong, Lin; Shen, De-Zhen

    2017-02-01

    Paper, as one of the most important information carriers, has contributed to the development and transmission of human civilization greatly. Meanwhile, a serious problem of environmental sustainable development caused by the production and utilization of paper has been resulted to modern society. Therefore, a simple and green route is urgently demanded to realize rewritable painting on paper. Herein, a simple route to rewritable painting on copy paper has been demonstrated by using eco-friendly ZnO nanoparticles (NPs) as fluorescent ink, and vinegar and soda that are frequently used in kitchen as erasing and neutralizing agents. Words or patterns written using the ZnO NPs as ink can be erased by vinegar vapour within five seconds, and after a neutralizing process in the ambient of soda vapour, the paper can be used for writing again. It is worth noting that the resolution and precision of the patterns produced via the above route degrade little after ten rewriting cycles, and the quality of the patterns produced using the ZnO NPs as ink fades little after being storage for several months, which promises the versatile potential applications of the rewriting route proposed in this paper.

  4. Single-molecule-sensitive fluorescence resonance energy transfer in freely-diffusing attoliter droplets

    NASA Astrophysics Data System (ADS)

    Rahmanseresht, Sheema; Milas, Peker; Ramos, Kieran P.; Gamari, Ben D.; Goldner, Lori S.

    2015-05-01

    Fluorescence resonance energy transfer (FRET) from individual, dye-labeled RNA molecules confined in freely-diffusing attoliter-volume aqueous droplets is carefully compared to FRET from unconfined RNA in solution. The use of freely-diffusing droplets is a remarkably simple and high-throughput technique that facilitates a substantial increase in signal-to-noise for single-molecular-pair FRET measurements. We show that there can be dramatic differences between FRET in solution and in droplets, which we attribute primarily to an altered pH in the confining environment. We also demonstrate that a sufficient concentration of a non-ionic surfactant mitigates this effect and restores FRET to its neutral-pH solution value. At low surfactant levels, even accounting for pH, we observe differences between the distribution of FRET values in solution and in droplets which remain unexplained. Our results will facilitate the use of nanoemulsion droplets as attoliter volume reactors for use in biophysical and biochemical assays, and also in applications such as protein crystallization or nanoparticle synthesis, where careful attention to the pH of the confined phase is required.

  5. Rewritable Painting Realized from Ambient-Sensitive Fluorescence of ZnO Nanoparticles.

    PubMed

    Liu, Kai-Kai; Shan, Chong-Xin; He, Gao-Hang; Wang, Ruo-Qiu; Dong, Lin; Shen, De-Zhen

    2017-02-07

    Paper, as one of the most important information carriers, has contributed to the development and transmission of human civilization greatly. Meanwhile, a serious problem of environmental sustainable development caused by the production and utilization of paper has been resulted to modern society. Therefore, a simple and green route is urgently demanded to realize rewritable painting on paper. Herein, a simple route to rewritable painting on copy paper has been demonstrated by using eco-friendly ZnO nanoparticles (NPs) as fluorescent ink, and vinegar and soda that are frequently used in kitchen as erasing and neutralizing agents. Words or patterns written using the ZnO NPs as ink can be erased by vinegar vapour within five seconds, and after a neutralizing process in the ambient of soda vapour, the paper can be used for writing again. It is worth noting that the resolution and precision of the patterns produced via the above route degrade little after ten rewriting cycles, and the quality of the patterns produced using the ZnO NPs as ink fades little after being storage for several months, which promises the versatile potential applications of the rewriting route proposed in this paper.

  6. Non-destructive trace element microanalysis of as-received cometary nucleus samples using synchrotron x ray fluorescence

    NASA Technical Reports Server (NTRS)

    Sutton, S. R.

    1989-01-01

    The Synchrotron X ray Fluorescence (SXRF) microprobe at the National Synchrotron Light Source (NSLS), Brookhaven National Laboratory, will be an excellent instrument for non-destructive trace element analyses of cometary nucleus samples. Trace element analyses of as-received cometary nucleus material will also be possible with this technique. Bulk analysis of relatively volatile elements will be important in establishing comet formation conditions. However, as demonstrated for meteorites, microanalyses of individual phases in their petrographic context are crucial in defining the histories of particular components in unequilibrated specimens. Perhaps most informative in comparing cometary material with meteorites will be the halogens and trace metals. In-situ, high spatial resolution microanalyses will be essential in establishing host phases for these elements and identifying terrestrial (collection/processing) overprints. The present SXRF microprobe is a simple, yet powerful, instrument in which specimens are excited with filtered, continuum synchrotron radiation from a bending magnet on a 2.5 GeV electron storage ring. A refrigerated cell will be constructed to permit analyses at low temperatures. The cell will consist essentially of an air tight housing with a cold stage. Kapton windows will be used to allow the incident synchrotron beam to enter the cell and fluorescent x rays to exit it. The cell will be either under vacuum or continuous purge by ultrapure helium during analyses. Several other improvements of the NSLS microprobe will be made prior to the cometary nucleus sample return mission that will greatly enhance the sensitivity of the technique.

  7. Colloidal graphene as a transducer in homogeneous fluorescence-based immunosensor for rapid and sensitive analysis of microcystin-LR.

    PubMed

    Liu, Meng; Zhao, Huimin; Chen, Shuo; Yu, Hongtao; Quan, Xie

    2012-11-20

    Herein, we reported the assembly of colloidal graphene (CG) and microcystin (MC)-LR-DNA bioconjugates to develop a homogeneous competitive fluorescence-based immunoassay for rapid and sensitive detection of MC-LR in water samples. Initially, the MC-LR-DNA probe was quickly adsorbed onto the CG surface through the strong noncovalent π-π stacking interactions and can be effectively quenched benefiting from the high quenching efficiency of CG. In contrast, the competitive binding of anti-MC-LR with MC-LR-DNA destroyed the graphene/MC-LR-DNA interaction, thus resulting in the restoration of fluorescence signal. This signal transduction mechanism made it possible for analysis of the target MC-LR. Taking advantage of the colloidal nature of the as-prepared graphene, the assay was carried out in homogeneous solution throughout, which avoided numerous immobilization, incubation, and washing steps that were necessary to traditional heterogeneous immunoassays, thereby reducing the whole assay time (within less than 35 min) and allowing a much better antigen-antibody interaction. Moreover, due to the direct competitive mode, the assay did not involve any antibody labeling or modification process, which would be beneficial to preserve the binding affinity of antigen-antibody. Under optimal conditions, the proposed immunosensor can be applied for quantitative analysis of MC-LR with a detection limit of 0.14 μg/L, which satisfied the World Health Organization (WHO) provisional guideline limit of 1 μg/L for MC-LR in drinking water, thus providing a powerful tool for rapid and sensitive monitoring of MC-LR in environmental samples.

  8. The ISAS Synchrotron Microprobe at DELTA

    SciTech Connect

    Bohlen, Alex von; Kraemer, Markus; Hergenroeder, Roland; Berges, Ulf

    2007-01-19

    Since 2004 ISAS operates a dipole beamline at the synchrotron radiation facility DELTA at University of Dortmund. Synchrotron radiation is used at this beamline as an excellent excitation source for X-ray fluorescence spectrometry (XRF). Among others, the high brilliance of the synchrotron radiation in contrast to conventional X-ray tubes, the strong polarization of the synchrotron radiation and the low divergence of the electron beam can be applied to XRF offering several advantages for spectroscopy. These outstanding features encouraged us to develop and operate a synchrotron radiation induced X-ray micro fluorescence probe connected to a wavelength dispersive spectrometer (SR-WDXRF). A relevant characteristic of such a device, namely, good lateral resolution at high spectral resolution can be applied for single spot-, line-scan and area map analyses of a variety of objects. The instrumentation of the SR-WDXRF and the performed experiments will be presented. Main task is the detection of light elements by their fluorescence K-lines and the specification of element compounds.

  9. One-pot synthesis of mesoporous structured ratiometric fluorescence molecularly imprinted sensor for highly sensitive detection of melamine from milk samples.

    PubMed

    Xu, Shoufang; Lu, Hongzhi

    2015-11-15

    A facile strategy was developed to prepare mesoporous structured ratiometric fluorescence molecularly imprinted sensor for highly sensitive and selective determination of melamine using CdTe QDs as target sensitive dye and hematoporphyrin as reference dyes. One-pot synthesis method was employed because it could simplify the imprinting process and shorten the experimental period. The as-prepared fluorescence MIPs sensor, which combined ratiometric fluorescence technique with mesoporous silica materials into one system, exhibited excellent selectivity and sensitivity. Under optimum conditions, these mesoporous structured ratiometric fluorescence MIP@QDs sensors showed detection limit as low as 38 nM, which was much lower than those non-mesoporous one. The recycling process was sustainable at least 10 times without obvious efficiency decrease. The feasibility of the developed method in real samples was successfully evaluated through the analysis of melamine in raw milk and milk powder samples with satisfactory recoveries of 92-101%. The developed method proposed in this work proved to be a convenient, rapid, reliable and practical way to prepared high sensitive and selective fluorescence sensors with potentially applicable for trace pollutants analysis in complicated samples.

  10. Secretory fluorescent protein, a secretion green fluorescent fusion protein with alkaline phosphatase activity as a sensitive and traceable reporter in baculovirus expression system.

    PubMed

    Teng, Chao-Yi; Wu, Tzong-Yuan

    2007-07-01

    The advantages of using traceable fluorescent protein (enhanced green fluorescent protein; EGFP) and a secretory alkaline phosphatase (SEAP) have been used to generate a reporter gene: the secretory fluorescent protein (SEFP). Sf21 cells, infected with the recombinant baculovirus containing the SEFP gene, revealed both traceable fluorescence and easily detectable alkaline phosphatase activity in the culture medium. The distribution of SEFP within the cells revealed that it was excluded from the nucleus, implying that the accumulation of SEFP in a secretory pathway, similar to that of the secretion signal-tagged FPs. Furthermore, the time- and dose-dependent release from the blockage of brefeldin A (BFA) confirmed that the secretion of SEFP was mediated by the secretion pathway and excluded leakage from viral infection. This SEFP reporter gene with traceable fluorescence and alkaline phosphatase activity may become a useful tool for studies on secretory protein production.

  11. "Turn-off" fluorescent sensor for highly sensitive and specific simultaneous recognition of 29 famous green teas based on quantum dots combined with chemometrics.

    PubMed

    Liu, Li; Fan, Yao; Fu, Haiyan; Chen, Feng; Ni, Chuang; Wang, Jinxing; Yin, Qiaobo; Mu, Qingling; Yang, Tianming; She, Yuanbin

    2017-04-22

    Fluorescent "turn-off" sensors based on water-soluble quantum dots (QDs) have drawn increasing attention owing to their unique properties such as high fluorescence quantum yields, chemical stability and low toxicity. In this work, a novel method based on the fluorescence "turn-off" model with water-soluble CdTe QDs as the fluorescent probes for differentiation of 29 different famous green teas is established. The fluorescence of the QDs can be quenched in different degrees in light of positions and intensities of the fluorescent peaks for the green teas. Subsequently, with aid of classic partial least square discriminant analysis (PLSDA), all the green teas can be discriminated with high sensitivity, specificity and a satisfactory recognition rate of 100% for training set and 98.3% for prediction set, respectively. Especially, the "turn-off" fluorescence PLSDA model based on second-order derivatives (2nd der) with reduced least complexity (LVs = 3) was the most effective one for modeling. Most importantly, we further demonstrated the established "turn-off" fluorescent sensor mode has several significant advantages and appealing properties over the conventional fluorescent method for large-class-number classification (LCNC) of green teas. This work is, to the best of our knowledge, the first report on the rapid and effective identification of so many kinds of famous green teas based on the "turn-off" model of QDs combined with chemometrics, which also implies other potential applications on complex LCNC classification system with weak fluorescence or even without fluorescence to achieve higher detective response and specificity.

  12. Searches for Microbial Cells with Fluorescence Loggers with Single-cell Sensitivity

    NASA Astrophysics Data System (ADS)

    Price, P. B.; Rohde, R. A.; Bay, R. C.

    2007-12-01

    Two known habitats for microbial metabolism in ice are surfaces of mineral grains and liquid veins along three- grain boundaries. Several problems suggest the need for a third habitat: veins usually contain toxic liquid; some microorganisms are too large to fit into a vein; veins may not be present at all depths; and the oxygen concentration in veins does not permit the coexistence of both strict anaerobes and aerobes in the same region. We show that a more general habitat avoids these problems. Isolated microbes frozen in ice and not in contact with a vein or grain can metabolize by redox reactions with dissolved small molecules diffusing through the ice lattice. The two requirements are that the gaseous reactants have sufficiently high equilibrium concentrations and diffusion coefficients to provide enough metabolic energy to repair macromolecular damage as it occurs. Molecules with less than ~6 atoms (e.g., H2, O2, N2¬, CO, CO2, CH4, H2S, NH3, HNO3, HCHO, and HCOOH) have values of diffusion coefficient D(T) that exceed ~10- 15 m2 s-1, which is sufficient to sustain microbial life in ice. For terrestrial environments, we show that there is an adequate supply of such molecules diffusing throughout deep glacial ice to sustain metabolism for millions of years. Our recent noninvasive observations of ice cores from GISP2 and WAIS Divide provide evidence for this habitat. Using scanning fluorimetry to map proteins (a proxy for cells) and F420 (a proxy for methanogens) in ice cores, we find isolated spikes of fluorescence consistent with as few as one microbial cell in a volume 0.16 microliter with the protein mapper and in 1.9 microliter with the methanogen mapper. With such precise localization one could use a nanomanipulator to extract single cells for molecular identification. Low- power, miniaturized versions of these instruments could search for single cells in subglacial lakes, Martian ice- rich permafrost, and Europan ice.

  13. U/Th dating by SHRIMP RG ion-microprobe mass spectrometry using single ion-exchange beads

    USGS Publications Warehouse

    Bischoff, J.L.; Wooden, J.; Murphy, F.; Williams, Ross W.

    2005-01-01

    We present a new analytical method for U-series isotopes using the SHRIMP RG (Sensitive High mass Resolution Ion MicroProbe) mass spectrometer that utilizes the preconcentration of the U-series isotopes from a sample onto a single ion-exchange bead. Ion-microprobe mass spectrometry is capable of producing Th ionization efficiencies in excess of 2%. Analytical precision is typically better than alpha spectroscopy, but not as good as thermal ionization mass spectroscopy (TIMS) and inductively coupled plasma multicollector mass spectrometry (ICP-MS). Like TIMS and ICP-MS the method allows analysis of small samples sizes, but also adds the advantage of rapidity of analysis. A major advantage of ion-microprobe analysis is that U and Th isotopes are analyzed in the same bead, simplifying the process of chemical separation. Analytical time on the instrument is ???60 min per sample, and a single instrument-loading can accommodate 15-20 samples to be analyzed in a 24-h day. An additional advantage is that the method allows multiple reanalyses of the same bead and that samples can be archived for reanalysis at a later time. Because the ion beam excavates a pit only a few ??m deep, the mount can later be repolished and reanalyzed numerous times. The method described of preconcentrating a low concentration sample onto a small conductive substrate to allow ion-microprobe mass spectrometry is potentially applicable to many other systems. Copyright ?? 2005 Elsevier Ltd.

  14. Elastic recoil detection analysis on the ANSTO heavy ion microprobe

    NASA Astrophysics Data System (ADS)

    Siegele, R.; Orlic, I.; Cohen, David D.

    2002-05-01

    The heavy ion microprobe at the Australian Nuclear Science and Technology Organisation is capable of focussing heavy ions with an ME/ q2 of up to 100 amu MeV. This makes the microprobe ideally suited for heavy ion elastic recoil detection analysis (ERDA). However, beam currents on a microprobe are usually very small, which requires a detection system with a large solid angle. We apply microbeam heavy ion ERDA using a large solid angle ΔE- E telescope with a gas ΔE detector to layered structures. We demonstrate the capability to measure oxygen and carbon with a lateral resolution of 20 μm, together with determination of the depth of the contamination in thin deposited layers.

  15. DNA-stabilized silver nanoclusters and carbon nanoparticles oxide: A sensitive platform for label-free fluorescence turn-on detection of HIV-DNA sequences.

    PubMed

    Ye, Yu-Dan; Xia, Li; Xu, Dang-Dang; Xing, Xiao-Jing; Pang, Dai-Wen; Tang, Hong-Wu

    2016-11-15

    Based on the remarkable difference between the interactions of carbon nanoparticles (CNPs) oxide with single-stranded DNA (ssDNA) and double-stranded DNA (dsDNA), and the fact that fluorescence of DNA-stabilized silver nanoclusters (AgNCs) can be quenched by CNPs oxide, DNA-functionalized AgNCs were applied as label-free fluorescence probes and a novel fluorescence resonance energy transfer (FRET) sensor was successfully constructed for the detection of human immunodeficiency virus (HIV) DNA sequences. CNPs oxide were prepared with the oxidation of candle soot, hence it is simple, time-saving and low-cost. The strategy of dual AgNCs probes was applied to improve the detection sensitivity by using dual- probe capturing the same target DNA in a sandwich mode and as the fluorescence donor, and using CNPs oxide as the acceptor. In the presence of target DNA, a dsDNA hybrid forms, leading to the desorption of the ssDNA-AgNCs probes from CNPs oxide, and the recovering of fluorescence of the AgNCs in a HIV-DNA concentration-dependent manner. The results show that HIV-DNA can be detected in the range of 1-50nM with a detection limit of 0.40nM in aqueous buffer. The method is simple, rapid and sensitive with no need of labeled fluorescent probes, and moreover, the design of fluorescent dual-probe makes full use of the excellent fluorescence property of AgNCs and further improves the detection sensitivity.

  16. Single-layer MnO2 nanosheets suppressed fluorescence of 7-hydroxycoumarin: mechanistic study and application for sensitive sensing of ascorbic acid in vivo.

    PubMed

    Zhai, Wanying; Wang, Chunxia; Yu, Ping; Wang, Yuexiang; Mao, Lanqun

    2014-12-16

    In this study, we systematically investigate the mechanism of single-layer MnO2 nanosheets suppressing fluorescence of 7-hydroxycoumarin and, based on this, demonstrate a new fluorescent method for in vivo sensing of ascorbic acid (AA) in rat brain. The mechanism for the fluorescence suppression is attributed to a combination of inner filter effect (IFE) and static quenching effect (SQE), which is different from those reported for the traditional two-dimensional nanosheets, and Förster resonant energy transfer (FRET) mechanism reported for MnO2 nanosheets. The combination of IFE and SQE leads to an exponential decay in fluorescence intensity of 7-hydroxycoumarin with increasing concentration of MnO2 nanosheets in solution. Such a property allows optimization of the concentration of MnO2 nanosheets in such a way that the addition of reductive analyte (e.g., AA) will to the greatest extent restore the MnO2 nanosheets-suppressed fluorescence of 7-hydroxycoumarin through the redox reaction between AA and MnO2 nanosheets. On the basis of this feature, we demonstrate a fluorescent method for in vivo sensing of AA in the cerebral systems with an improved sensitivity. Compared with the turn-on fluorescent method through first decreasing the fluorescence to the lowest level by adding concentrated MnO2 nanosheets, the method demonstrated here possesses a higher sensitivity, lower limit of detection, and wider linear range. Upon the use of ascorbate oxidase to achieve the selectivity for AA, the turn-on fluorescence method demonstrated here can be used for in vivo sensing of AA in a simple but reliable way.

  17. A new hydroxynaphthyl benzothiazole derived fluorescent probe for highly selective and sensitive Cu2 + detection

    NASA Astrophysics Data System (ADS)

    Tang, Lijun; He, Ping; Zhong, Keli; Hou, Shuhua; Bian, Yanjiang

    2016-12-01

    A new reactive probe, 1-(benzo[d]thiazol-2-yl)naphthalen-2-yl-picolinate (BTNP), was designed and synthesized. BTNP acts as a highly selective probe to Cu2 + in DMSO/H2O (7/3, v/v, Tris-HCl 10 mM, pH = 7.4) solution based on Cu2 + catalyzed hydrolysis of the picolinate ester moiety in BTNP, which leads to the formation of an ESIPT active product with dual wavelength emission enhancement. The probe also possesses the advantages of simple synthesis, rapid response and high sensitivity. The pseudo-first-order reaction rate constant was calculated to be 0.205 min- 1. Moreover, application of BTNP to Cu2 + detection in living cells and real water samples was also explored.

  18. Imaging Synaptic Vesicle Exocytosis-Endocytosis with pH-Sensitive Fluorescent Proteins.

    PubMed

    Afuwape, Olusoji A T; Kavalali, Ege T

    2016-01-01

    The introduction of pHluorin, a pH-sensitive GFP, by Miesenbock and colleagues provided a versatile tool to studies of vesicle trafficking, in particular synaptic vesicle exocytosis and endocytosis. By tagging pHluorin to the luminal region of the synaptic vesicular protein synaptobrevin (also called VAMP, vesicle-associated membrane protein) or other synaptic vesicle-specific proteins such as the vesicular glutamate transporter-1, we are able to directly track synaptic vesicle endocytosis in response to stimuli in a molecularly specific manner. Here, we describe the process of imaging synaptic vesicle endocytosis in response to extracellular stimulation in dissociated neuronal cultures of hippocampal neurons obtained from rats-also applicable to mice-using pHluorin-tagged vesicular glutamate transporter-1 as a reporter.

  19. Ultra-sensitive detection of kanamycin for food safety using a reduced graphene oxide-based fluorescent aptasensor

    NASA Astrophysics Data System (ADS)

    Ha, Na-Reum; Jung, In-Pil; La, Im-Joung; Jung, Ho-Sup; Yoon, Moon-Young

    2017-01-01

    Overuse of antibiotics has caused serious problems, such as appearance of super bacteria, whose accumulation in the human body through the food chain is a concern. Kanamycin is a common antibiotic used to treat diverse infections; however, residual kanamycin can cause many side effects in humans. Thus, development of an ultra-sensitive, precise, and simple detection system for residual kanamycin in food products is urgently needed for food safety. In this study, we identified kanamycin-binding aptamers via a new screening method, and truncated variants were analyzed for optimization of the minimal sequence required for target binding. We found various aptamers with high binding affinity from 34.7 to 669 nanomolar Kdapp values with good specificity against kanamycin. Furthermore, we developed a reduced graphene oxide (RGO)-based fluorescent aptasensor for kanamycin detection. In this system, kanamycin was detected at a concentration as low as 1 pM (582.6 fg/mL). In addition, this method could detect kanamycin accurately in kanamycin-spiked blood serum and milk samples. Consequently, this simple, rapid, and sensitive kanamycin detection system with newly structural and functional analysis aptamer exhibits outstanding detection compared to previous methods and provides a new possibility for point of care testing and food safety.

  20. Ultra-sensitive detection of kanamycin for food safety using a reduced graphene oxide-based fluorescent aptasensor

    PubMed Central

    Ha, Na-Reum; Jung, In-Pil; La, Im-Joung; Jung, Ho-Sup; Yoon, Moon-Young

    2017-01-01

    Overuse of antibiotics has caused serious problems, such as appearance of super bacteria, whose accumulation in the human body through the food chain is a concern. Kanamycin is a common antibiotic used to treat diverse infections; however, residual kanamycin can cause many side effects in humans. Thus, development of an ultra-sensitive, precise, and simple detection system for residual kanamycin in food products is urgently needed for food safety. In this study, we identified kanamycin-binding aptamers via a new screening method, and truncated variants were analyzed for optimization of the minimal sequence required for target binding. We found various aptamers with high binding affinity from 34.7 to 669 nanomolar Kdapp values with good specificity against kanamycin. Furthermore, we developed a reduced graphene oxide (RGO)-based fluorescent aptasensor for kanamycin detection. In this system, kanamycin was detected at a concentration as low as 1 pM (582.6 fg/mL). In addition, this method could detect kanamycin accurately in kanamycin-spiked blood serum and milk samples. Consequently, this simple, rapid, and sensitive kanamycin detection system with newly structural and functional analysis aptamer exhibits outstanding detection compared to previous methods and provides a new possibility for point of care testing and food safety. PMID:28054670

  1. Sensitive detection of proteins in polyacrylamide gel via isatoic anhydride derivatization: Introduction of a low-cost fluorescent prelabeling procedure.

    PubMed

    Asadollahi, Kazem; Rafiee, Saharnaz; Riazi, Gholamhossein

    2016-10-01

    Here, we introduce isatoic anhydride as a sensitive and commodious fluorescent prelabel for detection of proteins in one-dimensional polyacrylamide gels. High reactivity of isatoic anhydride with nucleophiles in mild alkaline environments makes it an appropriate tag for labeling of biomolecules. In this study, we show that preelectrophoresis labeling of proteins with isatoic anhydride for few minutes at room temperature allows detection of 2-4 ng of standard proteins, BSA and lysozyme, per band. Proteins were successfully labeled in the presence of a wide range of common biological reagents and in crude cell extract. The labeled proteins have the same electrophoretic migration in comparison to unlabeled proteins; however the application of saturation labeling method results in slight band broadening. Compatibility of the method with downstream processes was assessed by tryptic digestion of labeled proteins and study of peptide mixture using gel electrophoresis which revealed partial digestion of labeled proteins due to lysine modification. The present procedure is sensitive, rapid, and inexpensive and is a promising alternative for current protein staining procedures, where downstream processes are not desired.

  2. Permethylated-β-Cyclodextrin Capped CdTe Quantum Dot and its Sensitive Fluorescence Analysis of Malachite Green.

    PubMed

    Cao, Yujuan; Wei, Jiongling; Wu, Wei; Wang, Song; Hu, Xiaogang; Yu, Ying

    2015-09-01

    In the present work, the CdTe quantum dots were covalently conjugated with permethylated-β-cyclodextrin (OMe-β-CD) using 1-ethyl-3-(3-dimethylaminopropyl) carbodiimide hydrochloride as cross-linking reagent. The obtained functional quantum dots (OMe-β-CD/QDs) showed highly luminescent, water solubility and photostability as well as good inclusion ability to malachite green. A sensitive fluorescence method was developed for the analysis of malachite green in different samples. The good linearity was 2.0 × 10(-7)-1.0 × 10(-5) mol/L and the limit of detect was 1.7 × 10(-8) mol/L. The recoveries for three environmental water samples were 92.0-108.2 % with relative standard deviation (RSD) of 0.24-1.87 %, while the recovery for the fish sample was 94.3 % with RSD of 1.04 %. The results showed that the present method was sensitive and convenient to determine malachite green in complex samples. Graphical Abstract The analytical mechanism of OMe-β-CD/QDs and its linear response to MG.

  3. Ultra-fast and sensitive detection of non-typhoidal Salmonella using microwave-accelerated metal-enhanced fluorescence ("MAMEF").

    PubMed

    Tennant, Sharon M; Zhang, Yongxia; Galen, James E; Geddes, Chris D; Levine, Myron M

    2011-04-08

    Certain serovars of Salmonella enterica subsp. enterica cause invasive disease (e.g., enteric fever, bacteremia, septicemia, meningitis, etc.) in humans and constitute a global public health problem. A rapid, sensitive diagnostic test is needed to allow prompt initiation of therapy in individual patients and for measuring disease burden at the population level. An innovative and promising new rapid diagnostic technique is microwave-accelerated metal-enhanced fluorescence (MAMEF). We have adapted this assay platform to detect the chromosomal oriC locus common to all Salmonella enterica subsp. enterica serovars. We have shown efficient lysis of biologically relevant concentrations of Salmonella spp. suspended in bacteriological media using microwave-induced lysis. Following lysis and DNA release, as little as 1 CFU of Salmonella in 1 ml of medium can be detected in <30 seconds. Furthermore the assay is sensitive and specific: it can detect oriC from Salmonella serovars Typhi, Paratyphi A, Paratyphi B, Paratyphi C, Typhimurium, Enteritidis and Choleraesuis but does not detect Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, Streptococcus pneumoniae, Haemophilus influenzae or Acinetobacter baumanii. We have also performed preliminary experiments using a synthetic Salmonella oriC oligonucleotide suspended in whole human blood and observed rapid detection when the sample was diluted 1:1 with PBS. These pre-clinical data encourage progress to the next step to detect Salmonella in blood (and other ordinarily sterile, clinically relevant body fluids).

  4. Microactuator fabricated by powder particle assemblage using microprobe technology

    NASA Astrophysics Data System (ADS)

    Konno, Takeshi; Egashira, Mitsuru; Shinya, Norio

    1997-06-01

    In the previous paper, preliminary research results on powder particle assemblage technique using a microprobe was reported. It was shown that the technique makes it possible to manipulate powder particles one by one, etch microscopically and weld the powder particle into a substrate or other powder particles. In this work, the welding mechanism of this method and metallurgical properties of welded parts were investigated, and micro- actuators were fabricated by means of powder particle assemblage technique using the microprobe. The results indicated the potentiality of this technique for application to assemblage of micro-machine and micro-devices.

  5. A fluorescent light-up aggregation-induced emission probe for screening gefitinib-sensitive non-small cell lung carcinoma.

    PubMed

    Hu, Yi; Shi, Leilei; Su, Yue; Zhang, Chuan; Jin, Xin; Zhu, Xinyuan

    2017-03-07

    Fluorescent light-up probes with aggregation-induced emission (AIE) characteristics have been focused on recently. In this report, a new fluorescent probe, namely, DEVD-TPE, which consisted of the substrate peptide Asp-Glu-Val-Asp (DEVD) and the AIE reporter group tetraphenylethene (TPE), was developed for detecting caspase-3 in living cells. In a slightly alkaline solution, the DEVD-TPE probe displayed almost no fluorescence owing to the dynamic rotation of the phenyl rings in solution. However, DEVD-TPE exhibited significant fluorescence when it was cleaved by caspase-3, as well as when the reporter group TPE underwent aggregation. The epidermal growth factor receptor (EGFR) inhibitor gefitinib was used for determining the screening efficacy of the probe for different non-small cell lung carcinoma (NSCLC) cell lines, namely, HCC827, A549 and H1650 cells. Cell proliferation and apoptosis assays indicated that the three cell lines had different sensitivities to gefitinib. The results of analysis by living-cell fluorescence imaging and flow cytometry were consistent with those of the cell proliferation and apoptosis assays. This demonstrated that our probe could detect caspase-3 in living cells, which confirmed the apoptosis of NSCLC cells. Furthermore, our probe indicated that gefitinib was more efficient against HCC827 cells than against the other two NSCLC cell lines. This report proves that the fluorescent probe DEVD-TPE is highly sensitive to caspase-3 and has potential prospects in the rapid screening of NSCLC.

  6. Developing a genetically encoded green fluorescent protein mutant for sensitive light-up fluorescent sensing and cellular imaging of Hg(II).

    PubMed

    Jiang, Tao; Guo, Daiping; Wang, Qian; Wu, Xin; Li, Zhao; Zheng, Zhenhua; Yin, Boyuan; Xia, Lin; Tang, Jixian; Luo, Wenxin; Xia, Ningshao; Jiang, Yunbao

    2015-05-30

    Hg(II) is well-known for quenching fluorescence in a distance dependent manner. Nevertheless, when we exposed the fluorophore of a green fluorescent protein (GFP) toward Hg(II), through H148C mutation, the GFP fluorescence could be "lighted up" by Hg(II) down to sub-nM level. The detection linear range is 0.5-3.0 nM for protein solutions at 8.0 nM. The GFPH148C protein displayed a promising selectivity toward Hg(II) and also the cellular imaging capacity. Spectra measurements suggested that the ground-state redistribution of protein contributed to the fluorescence enhancement, which was found not limited to Hg(II), and thus presented an opening for building a pool of GFP-based chemosensors toward other heavy metal ions.

  7. Photochemical studies of a fluorescent chlorophyll catabolite--source of bright blue fluorescence in plant tissue and efficient sensitizer of singlet oxygen.

    PubMed

    Jockusch, Steffen; Turro, Nicholas J; Banala, Srinivas; Kräutler, Bernhard

    2014-02-01

    Fluorescent chlorophyll catabolites (FCCs) are fleeting intermediates of chlorophyll breakdown, which is seen as an enzyme controlled detoxification process of the chlorophylls in plants. However, some plants accumulate large amounts of persistent FCCs, such as in senescent leaves and in peels of yellow bananas. The photophysical properties of such a persistent FCC (Me-sFCC) were investigated in detail. FCCs absorb in the near UV spectral region and show blue fluorescence (max at 437 nm). The Me-sFCC fluorescence had a quantum yield of 0.21 (lifetime 1.6 ns). Photoexcited Me-sFCC intersystem crosses into the triplet state (quantum yield 0.6) and generates efficiently singlet oxygen (quantum yield 0.59). The efficient generation of singlet oxygen makes fluorescent chlorophyll catabolites phototoxic, but might also be useful as a (stress) signal and for defense of the plant tissue against infection by pathogens.

  8. A quantitative protocol for dynamic measurements of protein interactions by Förster resonance energy transfer-sensitized fluorescence emission

    PubMed Central

    Elder, A.D.; Domin, A.; Kaminski Schierle, G.S.; Lindon, C.; Pines, J.; Esposito, A.; Kaminski, C.F.

    2008-01-01

    Fluorescence detection of acceptor molecules sensitized by Förster resonance energy transfer (FRET) is a powerful method to study protein interactions in living cells. The method requires correction for donor spectral bleed-through and acceptor cross-excitation as well as the correct normalization of signals to account for varying fluorophore concentrations and imaging parameters. In this paper, we review different methods for FRET signal normalization and then present a rigorous model for sensitized emission measurements, which is both intuitive to understand and practical to apply. The method is validated by comparison with the acceptor photobleaching and donor lifetime-imaging techniques in live cell samples containing EYFP and ECFP tandem constructs exhibiting known amounts of FRET. By varying the stoichiometry of interaction in a controlled fashion, we show that information on the fractions of interacting donors and acceptors can be recovered. Furthermore, the method is tested by performing measurements on different microscopy platforms in both widefield and confocal imaging modes to show that signals recovered under different imaging conditions are in quantitative agreement. Finally, the method is applied in the study of dynamic interactions in the cyclin–cdk family of proteins in live cells. By normalizing the obtained signals for both acceptor and donor concentrations and using a FRET exhibiting control construct for calibration, stoichiometric changes in these interactions could be visualized in real time. The paper is written to be of practical use to researchers interested in performing sensitized emission measurements. The correct interpretation of the retrieved signals in a biological context is emphasized, and guidelines are given for the practical application of the developed algorithms.

  9. Silver Nanoparticle-Based Fluorescence-Quenching Lateral Flow Immunoassay for Sensitive Detection of Ochratoxin A in Grape Juice and Wine

    PubMed Central

    Jiang, Hu; Li, Xiangmin; Xiong, Ying; Pei, Ke; Nie, Lijuan; Xiong, Yonghua

    2017-01-01

    A silver nanoparticle (AgNP)-based fluorescence-quenching lateral flow immunoassay with competitive format (cLFIA) was developed for sensitive detection of ochratoxin A (OTA) in grape juice and wine samples in the present study. The Ru(phen)32+-doped silica nanoparticles (RuNPs) were sprayed on the test and control line zones as background fluorescence signals. The AgNPs were designed as the fluorescence quenchers of RuNPs because they can block the exciting light transferring to the RuNP molecules. The proposed method exhibited high sensitivity for OTA detection, with a detection limit of 0.06 µg/L under optimized conditions. The method also exhibited a good linear range for OTA quantitative analysis from 0.08 µg/L to 5.0 µg/L. The reliability of the fluorescence-quenching cLFIA method was evaluated through analysis of the OTA-spiked red grape wine and juice samples. The average recoveries ranged from 88.0% to 110.0% in red grape wine and from 92.0% to 110.0% in grape juice. Meanwhile, less than a 10% coefficient variation indicated an acceptable precision of the cLFIA method. In summary, the new AgNP-based fluorescence-quenching cLFIA is a simple, rapid, sensitive, and accurate method for quantitative detection of OTA in grape juice and wine or other foodstuffs. PMID:28264472

  10. A turn-off fluorescent biosensor for the rapid and sensitive detection of uranyl ion based on molybdenum disulfide nanosheets and specific DNAzyme

    NASA Astrophysics Data System (ADS)

    Zhang, HongYan; Ruan, YaJuan; Lin, Ling; Lin, Minggui; Zeng, Xiaoxue; Xi, Zhiming; Fu, FengFu

    2015-07-01

    A novel fluorescent biosensor for detecting uranyl ion (UO22+) in aqueous environment has been developed based on the specific recognition of DNAzyme and the fluorescence quenching ability of molybdenum disulfide (MoS2) nanosheets. The DNAzyme contains a DNA enzyme strand and a 6-carboxylfluorescein (FAM)-labeled DNA substrate strand. We demonstrated that MoS2 nanosheets have low affinity to the substrate-enzyme complex DNAzyme. Whereas, in the presence of UO22+, UO22+ can specifically cleave DNAzyme to release FAM-labeled single-strand DNA and the released FAM-labeled single-strand DNA can be firmly adsorbed on the surface of MoS2 nanosheets, which resulted in an obvious decrease of fluorescence intensity. This provided a sensing platform for the rapid, simple and sensitive fluorescent detection of UO22+. By using the sensing platform, a sensitive and selective fluorescent method for the rapid detection of UO22+ has been developed. In comparison with previous biosensor, the proposed method has obvious analytical advantage such as relatively high sensitivity and good stability, short analytical time and low cost. It can be used to detect as low as 2.14 nM of UO22+ in aqueous environment with a recovery of 96-102% and a RSD < 5% (n = 6). The success of this study provides a promising alternative for the rapid and on-site detection of UO22+ in environmental monitoring.

  11. Silver Nanoparticle-Based Fluorescence-Quenching Lateral Flow Immunoassay for Sensitive Detection of Ochratoxin A in Grape Juice and Wine.

    PubMed

    Jiang, Hu; Li, Xiangmin; Xiong, Ying; Pei, Ke; Nie, Lijuan; Xiong, Yonghua

    2017-02-28

    A silver nanoparticle (AgNP)-based fluorescence-quenching lateral flow immunoassay with competitive format (cLFIA) was developed for sensitive detection of ochratoxin A (OTA) in grape juice and wine samples in the present study. The Ru(phen) 3 2 + -doped silica nanoparticles (RuNPs) were sprayed on the test and control line zones as background fluorescence signals. The AgNPs were designed as the fluorescence quenchers of RuNPs because they can block the exciting light transferring to the RuNP molecules. The proposed method exhibited high sensitivity for OTA detection, with a detection limit of 0.06 µg/L under optimized conditions. The method also exhibited a good linear range for OTA quantitative analysis from 0.08 µg/L to 5.0 µg/L. The reliability of the fluorescence-quenching cLFIA method was evaluated through analysis of the OTA-spiked red grape wine and juice samples. The average recoveries ranged from 88.0% to 110.0% in red grape wine and from 92.0% to 110.0% in grape juice. Meanwhile, less than a 10% coefficient variation indicated an acceptable precision of the cLFIA method. In summary, the new AgNP-based fluorescence-quenching cLFIA is a simple, rapid, sensitive, and accurate method for quantitative detection of OTA in grape juice and wine or other foodstuffs.

  12. 6-N,N-dimethylamino-2,3-naphthalimide: a new environment-sensitive fluorescent probe in delta- and mu-selective opioid peptides.

    PubMed

    Vázquez, M Eugenio; Blanco, Juan B; Salvadori, Severo; Trapella, Claudio; Argazzi, Roberto; Bryant, Sharon D; Jinsmaa, Yunden; Lazarus, Lawrence H; Negri, Lucia; Giannini, Elisa; Lattanzi, Roberta; Colucci, Mariantonella; Balboni, Gianfranco

    2006-06-15

    A new environment-sensitive fluorophore, 6-N,N-(dimethylamino)-2,3-naphthalimide (6DMN) was introduced in the delta-selective opioid peptide agonist H-Dmt-Tic-Glu-NH(2) and in the mu-selective opioid peptide agonist endomorphin-2 (H-Tyr-Pro-Phe-Phe-NH(2)). Environment-sensitive fluorophores are a special class of chromophores that generally exhibit a low quantum yield in aqueous solution but become highly fluorescent in nonpolar solvents or when bound to hydrophobic sites in proteins or membranes. New fluorescent delta-selective irreversible antagonists (H-Dmt-Tic-Glu-NH-(CH(2))(5)-CO-Dap(6DMN)-NH(2) (1) and H-Dmt-Tic-Glu-Dap(6DMN)-NH(2) (2)) were identified as potential fluorescent probes showing good properties for use in studies of distribution and internalization of delta receptors by confocal laser scanning microscopy.

  13. A label-free fluorescence DNA probe based on ligation reaction with quadruplex formation for highly sensitive and selective detection of nicotinamide adenine dinucleotide.

    PubMed

    Zhao, Jingjin; Zhang, Liangliang; Jiang, Jianhui; Shen, Guoli; Yu, Ruqin

    2012-05-11

    A simple label-free fluorescent sensing scheme for sensitive and selective detection of nicotinamide adenine dinucleotide (NAD(+)) has been developed based on DNA ligation reaction with ligand-responsive quadruplex formation. This approach can detect 0.5 nM NAD(+) with high selectivity against other NAD(+) analogs.

  14. An ultra-sensitive monoclonal antibody-based fluorescent microsphere immunochromatographic test strip assay for detecting aflatoxin M1 in milk

    Technology Transfer Automated Retrieval System (TEKTRAN)

    A rapid lateral flow fluorescent microspheres immunochromatography test strip (FMs-ICTS) has been developed for the detection of aflatoxin M1 (AFM1) residues in milk. For this purpose, an ultra-sensitive anti-AFM1 monoclonal antibody (MAb) 1D3 was prepared and identified. The IC50 value of the MA...

  15. Development of a pH sensor based on a nanostructured filter adding pH-sensitive fluorescent dye for detecting acetic acid in photovoltaic modules

    NASA Astrophysics Data System (ADS)

    Asaka, Takashi; Itayama, Tomohiro; Nagasaki, Hideaki; Iwami, Kentaro; Yamamoto, Chizuko; Hara, Yukiko; Masuda, Atsushi; Umeda, Norihiro

    2015-08-01

    Acetic acid formed via the hydrolysis of ethylene vinyl acetate (EVA) as an encapsulant in photovoltaic (PV) modules causes a decrease in the conversion efficiency of such modules by grid corrosion. Here, a nondestructive and simple optical method for evaluating the condition of PV modules is proposed. This method uses a dual-wavelength pH-sensitive fluorescent dye to detect acetic acid in PV modules using a change in pH. The change in pH induced by the formation of acetic acid is detected by the change in the ratio of the fluorescent intensities of two peaks of the dye. A pH-sensitive fluorescent dye showed sensitivity for small amounts of acetic acid such as that produced from EVA. Furthermore, a membrane filter dyed with a pH-sensitive fluorescent dye was confirmed to detect acetic acid in aged EVA after a damp-heat test (85 °C, 85%) for 5000 h in PV modules.

  16. A thermal microprobe fabricated with wafer-stage processing

    NASA Astrophysics Data System (ADS)

    Zhang, Yongxia; Zhang, Yanwei; Blaser, Juliana; Sriram, T. S.; Enver, Ahsan; Marcus, R. B.

    1998-05-01

    A thermal microprobe has been designed and built for high resolution temperature sensing. The thermal sensor is a thin-film thermocouple junction at the tip of an atomic force microprobe (AFM) silicon probe needle. Only wafer-stage processing steps are used for the fabrication. For high resolution temperature sensing it is essential that the junction be confined to a short distance at the AFM tip. This confinement is achieved by a controlled photoresist coating process. Experiment prototypes have been made with an Au/Pd junction confined to within 0.5 μm of the tip, with the two metals separated elsewhere by a thin insulating oxide layer. Processing begins with double-polished, n-type, 4 in. diameter, 300-μm-thick silicon wafers. Atomically sharp probe tips are formed by a combination of dry and wet chemical etching, and oxidation sharpening. The metal layers are sputtering deposited and the cantilevers are released by a combination of KOH and dry etching. A resistively heated calibration device was made for temperature calibration of the thermal microprobe over the temperature range 25-110 °C. Over this range the thermal outputs of two microprobes are 4.5 and 5.6 μV/K and is linear. Thermal and topographical images are also obtained from a heated tungsten thin film fuse.

  17. A TiS2 nanosheet enhanced fluorescence polarization biosensor for ultra-sensitive detection of biomolecules

    NASA Astrophysics Data System (ADS)

    Li, Xiang; Ding, Xuelian; Li, Yongfang; Wang, Linsong; Fan, Jing

    2016-05-01

    Development of new strategies for the sensitive and selective detection of ultra-low concentrations of specific cancer markers is of great importance for assessing cancer therapeutics due to its crucial role in early clinical diagnoses and biomedical applications. In this work, we have developed two types of fluorescence polarization (FP) amplification assay strategies for the detection of biomolecules by using TiS2 as a FP enhancer and Zn2+-dependent self-hydrolyzing deoxyribozymes as catalysts to realize enzyme-catalyzed target-recycling signal amplification. One approach is based on the terminal protection of small-molecule-linked DNA, in which biomolecular binding to small molecules in DNA-small-molecule chimeras can protect the conjugated DNA from degradation by exonuclease I (Exo I); the other approach is based on the terminal protection of biomolecular bound aptamer DNA, in which biomolecules directly bound to the single strand aptamer DNA can protect the ssDNA from degradation by Exo I. We select folate receptor (FR) and thrombin (Tb) as model analytes to verify the current concept. It is shown that under optimized conditions, our strategies exhibit high sensitivity and selectivity for the quantification of FR and Tb with low detection limits (0.003 ng mL-1 and 0.01 pM, respectively). Additionally, this strategy is a simple ``mix and detect'' approach, and does not require any separation steps. This biosensor is also utilized in the analysis of real biological samples, the results agree well with those obtained by the enzyme-linked immunosorbent assay (ELISA).Development of new strategies for the sensitive and selective detection of ultra-low concentrations of specific cancer markers is of great importance for assessing cancer therapeutics due to its crucial role in early clinical diagnoses and biomedical applications. In this work, we have developed two types of fluorescence polarization (FP) amplification assay strategies for the detection of biomolecules

  18. Sensitive detection and quantification of gliadin contamination in gluten-free food with immunomagnetic beads based liposomal fluorescence immunoassay.

    PubMed

    Chu, Pei-Tzu; Wen, Hsiao-Wei

    2013-07-17

    Gliadin from wheat is a common food allergen that can induce baker's asthma, wheat-dependent exercise-induced anaphylaxis, atopic dermatitis, and celiac disease. This gliadin assay focuses on rapidly screen and check for gluten contamination in raw materials and in the gluten-free food production process, not only for wheat-sensitive patients but also for the industries producing gluten-free foodstuffs. The developed assay incorporates the use of anti-gliadin antibody-conjugated immunomagnetic beads (IMBs) to capture the gliadin in samples and fluorescent dyes-loaded immunoliposomal nanovesicles (IMLNs) to produce and enhance the detection signal. Hence, a sandwich complex is formed as "IMBs-gliadin-IMLNs". Experimental results indicate that this detection platform exhibits good sensitivity for gliadin with a detection limit as low as 0.6 μg mL(-1) of gliadin; as the polyclonal antibody showed slight cross-reactions with barley and rye. Excellent recovery rates were found ranging from 83.5 to 102.6% as testing the spiked samples. Moreover, the CV (%) of intra- and inter-assay of this developed assay are 4.8-10.6% and 3.5-9.9%, respectively. Based on a parallel analysis of twenty food samples, the results of this developed assay provide a good consistency with those of an AOAC-approved ELISA kit without any false-negative results. The proposed assay method is thus a highly promising alternative method for detecting the contamination of gliadin in the food industry.

  19. investigating acid production by Streptococcus mutans with a surface-displayed pH-sensitive green fluorescent protein.

    PubMed

    Guo, Lihong; Hu, Wei; He, Xuesong; Lux, Renate; McLean, Jeff; Shi, Wenyuan

    2013-01-01

    Acidogenicity and aciduricity are the main virulence factors of the cavity-causing bacterium Streptococcus mutans. Monitoring at the individual cell level the temporal and spatial distribution of acid produced by this important oral pathogen is central for our understanding of these key virulence factors especially when S. mutans resides in multi-species microbial communities. In this study, we explored the application of pH-sensitive green fluorescent proteins (pHluorins) to investigate these important features. Ecliptic pHluorin was functionally displayed on the cell surface of S. mutans as a fusion protein with SpaP. The resulting strain (O87) was used to monitor temporal and spatial pH changes in the microenvironment of S. mutans cells under both planktonic and biofilm conditions. Using strain O87, we revealed a rapid pH drop in the microenviroment of S. mutans microcolonies prior to the decrease in the macro-environment pH following sucrose fermentation. Meanwhile, a non-uniform pH distribution was observed within S. mutans biofilms, reflecting differences in microbial metabolic activity. Furthermore, strain O87 was successfully used to monitor the S. mutans acid production profiles within dual- and multispecies oral biofilms. Based on these findings, the ecliptic pHluorin allows us to investigate in vivo and in situ acid production and distribution by the cariogenic species S. mutans.

  20. Peroxyoxalate chemiluminescence detection for the highly sensitive determination of fluorescence-labeled chlorpheniramine with Suzuki coupling reaction.

    PubMed

    Adutwum, Lawrence Asamoah; Kishikawa, Naoya; Ohyama, Kaname; Harada, Shiro; Nakashima, Kenichiro; Kuroda, Naotaka

    2010-09-01

    A sensitive and selective high performance liquid chromatography-peroxyoxalate chemiluminescence (PO-CL) method has been developed for the simultaneous determination of chlorpheniramine (CPA) and monodesmethyl chlorpheniramine (MDCPA) in human serum. The method combines fluorescent labeling with 4-(4,5-diphenyl-1H-imidazole-2-yl)phenyl boronic acid using Suzuki coupling reaction with PO-CL detection. CPA and MDCPA were extracted from human serum by liquid-liquid extraction with n-hexane. Excess labeling reagent, which interfered with trace level determination of analytes, was removed by solid-phase extraction using a C18 cartridge. Separation of derivatives of both analytes was achieved isocratically on a silica column with a mixture of acetonitrile and 60 mM imidazole-HNO(3) buffer (pH 7.2; 85:15, v/v) containing 0.015% triethylamine. The proposed method exhibited a good linearity with a correlation coefficient of 0.999 for CPA and MDCPA within the concentration range of 0.5-100 ng/mL. The limits of detection (S/N = 3) were 0.14 and 0.16 ng/mL for CPA and MDCPA, respectively. Using the proposed method, CPA could be selectively determined in human serum after oral administration.

  1. A highly sensitive fluorescent indicator dye for calcium imaging of neural activity in vitro and in vivo.

    PubMed

    Tada, Mayumi; Takeuchi, Atsuya; Hashizume, Miki; Kitamura, Kazuo; Kano, Masanobu

    2014-06-01

    Calcium imaging of individual neurons is widely used for monitoring their activity in vitro and in vivo. Synthetic fluorescent calcium indicator dyes are commonly used, but the resulting calcium signals sometimes suffer from a low signal-to-noise ratio (SNR). Therefore, it is difficult to detect signals caused by single action potentials (APs) particularly from neurons in vivo. Here we showed that a recently developed calcium indicator dye, Cal-520, is sufficiently sensitive to reliably detect single APs both in vitro and in vivo. In neocortical neurons, calcium signals were linearly correlated with the number of APs, and the SNR was > 6 for in vitro slice preparations and > 1.6 for in vivo anesthetised mice. In cerebellar Purkinje cells, dendritic calcium transients evoked by climbing fiber inputs were clearly observed in anesthetised mice with a high SNR and fast decay time. These characteristics of Cal-520 are a great advantage over those of Oregon Green BAPTA-1, the most commonly used calcium indicator dye, for monitoring the activity of individual neurons both in vitro and in vivo.

  2. Fluorescence detection of white-beam X-ray absorption anisotropy: towards element-sensitive projections of local atomic structure

    PubMed Central

    Korecki, P.; Tolkiehn, M.; Dąbrowski, K. M.; Novikov, D. V.

    2011-01-01

    Projections of the atomic structure around Nb atoms in a LiNbO3 single crystal were obtained from a white-beam X-ray absorption anisotropy (XAA) pattern detected using Nb K fluorescence. This kind of anisotropy results from the interference of X-rays inside a sample and, owing to the short coherence length of a white beam, is visible only at small angles around interatomic directions. Consequently, the main features of the recorded XAA corresponded to distorted real-space projections of dense-packed atomic planes and atomic rows. A quantitative analysis of XAA was carried out using a wavelet transform and allowed well resolved projections of Nb atoms to be obtained up to distances of 10 Å. The signal of nearest O atoms was detected indirectly by a comparison with model calculations. The measurement of white-beam XAA using characteristic radiation indicates the possibility of obtaining element-sensitive projections of the local atomic structure in more complex samples. PMID:21997909

  3. Sensitive mRNA detection by fluorescence in situ hybridization using horseradish peroxidase-labeled oligodeoxynucleotides and tyramide signal amplification.

    PubMed

    van de Corput, M P; Dirks, R W; van Gijlswijk, R P; van Binnendijk, E; Hattinger, C M; de Paus, R A; Landegent, J E; Raap, A K

    1998-11-01

    With the ongoing progress in human genome projects, many genes are discovered whose function and/or expression pattern are not known. Most of these genes are expressed in relatively low abundance compared to housekeeping genes such as elongation factor-1alpha and beta-actin. Gene expression is studied by Northern blot assays or by semiquantitative PCR methods. Another method is the visualization of transcripts in tissue or cell cultures by fluorescence in situ hybridization (FISH). However, for low-abundance RNA detection, this method is hampered by its limited detection sensitivity and by the interference of background signals with specific hybridization signals. Background signals are introduced by nonspecific hybridization of probe sequences or nonspecific binding of antibodies used for visualization. To eliminate background signals derived from both sources and to benefit from the peroxidase-driven tyramide signal amplification (TSA), we directly conjugated horseradish peroxidase (HRP) to oligodeoxynucleotides (ODNs) and used these probes to study in the bladder cancer cell line 5637 the expression of various cytokine genes which, according to Northern hybridization and reverse transcriptase-polymerase chain reaction (RT-PCR) assays, are expressed at levels up to 10,000-fold less than abundantly expressed housekeeping genes. The results show that reduction of probe complexity and the limited use of immunocytochemical detection layers strongly reduces noise signals derived from nonspecific binding of nucleic acid probe and antibodies. The use of the HRP-ODNs in combination with TSA allowed detection of low-abundance cytokine mRNAs by FISH.

  4. Non-redox modulated fluorescence strategy for sensitive and selective ascorbic acid detection with highly photoluminescent nitrogen-doped carbon nanoparticles via solid-state synthesis.

    PubMed

    Zhu, Xiaohua; Zhao, Tingbi; Nie, Zhou; Liu, Yang; Yao, Shouzhuo

    2015-08-18

    Highly photoluminescent nitrogen-doped carbon nanoparticles (N-CNPs) were prepared by a simple and green route employing sodium alginate as a carbon source and tryptophan as both a nitrogen source and a functional monomer. The as-synthesized N-CNPs exhibited excellent water solubility and biocompatibility with a fluorescence quantum yield of 47.9%. The fluorescence of the N-CNPs was intensively suppressed by the addition of ascorbic acid (AA). The mechanism of the fluorescence suppression of the N-CNPs was investigated, and the synergistic action of the inner filter effect (IFE) and the static quenching effect (SQE) contributed to the intensive fluorescence suppression, which was different from those reported for the traditional redox-based fluorescent probes. Owing to the spatial effect and hydrogen bond between the AA and the groups on the N-CNP surface, excellent sensitivity and selectivity for AA detecting was obtained in a wide linear relationship from 0.2 μM to 150 μM. The detection limit was as low as 50 nM (signal-to-noise ratio of 3). The proposed sensing systems also represented excellent sensitivity and selectivity for AA analysis in human biological fluids, providing a valuable platform for AA sensing in clinic diagnostic and drug screening.

  5. Sensitive fluorescent assay for copper (II) determination in aqueous solution using copper-specific ssDNA and Sybr Green I.

    PubMed

    Zhan, Shenshan; Xu, Hanchu; Zhang, Weilin; Zhan, Xuejia; Wu, Yuangen; Wang, Lumei; Zhou, Pei

    2015-09-01

    This paper reports a fluorescent turn-off assay for sensitive detection of Cu(2+) in an aqueous solution by using a copper-specific ssDNA Cu100 and Sybr Green I. By monitoring the fluorescence changes arose from different interactions of Sybr Green I with Cu100 and Cu100/Cu(2+) complex, the Cu(2+) could be linearly detected from 5.57 to 250 ppb, with a detection limit of 5.57 ppb. The feasibility of this assay was demonstrated by detecting Cu(2+) in certified reference materials and spiked water samples with satisfactory results.

  6. Use of selenium to detect mercury in water and cells: an enhancement of the sensitivity and specificity of a seleno fluorescent probe.

    PubMed

    Tang, Bo; Ding, Baiyu; Xu, Kehua; Tong, Lili

    2009-01-01

    Seleno fluorescent probe: An organoselenium fluorescent probe (FSe-1) for mercury was designed based on the irreversible deselenation mechanism. FSe-1 exhibits an ultrahigh selectivity and sensitivity for Hg(2+) detection only for reactive selenium atom sites, due the strong affinity between Se and Hg. Furthermore, the new probe has been successfully used for imaging mercury ions in RAW 264.7 cells (a mouse macrophage cell line; see figure).Inspired by the antitoxic function of selenium towards heavy-metal ions, we designed an organoselenium fluorescent probe (FSe-1) for mercury. The reaction of FSe-1 and Hg(2+) is an irreversible deselenation mechanism based on the selenophilic character of mercury. FSe-1 exhibits an ultrahigh selectivity and sensitivity for Hg(2+) detection only for reactive selenium atom sites due to the strong affinity between Se and Hg. The experimental results proved that FSe-1 was selective for Hg(2+) ions over other relevant metal ions and bioanalytes, and also showed an enhancement in sensitivity of up to 1.0 nM, which is lower than the current Environmental Protection Agency standard for drinking water. Furthermore, the new probe has been successfully applied to the imaging of mercury ions in RAW 264.7 cells (a mouse macrophage cell line) with high sensitivity and selectivity.

  7. Analysis of Light Gathering Abilities of Dynamically Solidified Micro-lenses, and Their Implementation to Improve Sensitivity of Fluorescent PCR Micro-detectors.

    PubMed

    Wu, Jian; Guo, Wei; Wang, Chunyan; Yu, Kuanxin; Chen, Tao; Li, Yinghui

    2015-06-01

    Fluorescent polymerase chain reaction (PCR) is becoming the preferred method of quantitative analysis due to its high specificity and sensitivity. We propose to use a new kind of micro-lens, dynamically solidified with optic glue, to improve the sensitivity of fluorescent PCR micro-detector. We developed light ray track equations for these lenses and used them to calculate relative light intensity distribution curve for stimulation lenses and illumination point probability distribution curve for detection lenses. We manufactured dynamically solidified micro-lenses using optic glue NOA61, and measured their light gathering ability. Lenses with radius/thickness (R/H) ratio of 4 reached light focusing ratio of 3.85 (stimulation lens) and photon collection efficiency of 0.86 (detection lens). We then used dynamically solidified lenses in PCR fluorescence micro-detector and analyzed their effect on the detector sensitivity. We showed that the use of dynamically solidified micro-lenses with R/H = 4 resulted in over 4.4-fold increased sensitivity of the detector.

  8. Hexa-BODIPY Linked-Triazole Based on a Cyclotriphosphazene Core as a Highly Selective and Sensitive Fluorescent Sensor for Fe(2+) Ions.

    PubMed

    Çetindere, Seda; Tümay, Süreyya O; Kılıç, Adem; Durmuş, Mahmut; Yeşilot, Serkan

    2016-07-01

    A new type of fluorescent chemosensor based on tethered hexa-borondipyrromethene cyclotriphosphazene platform (HBTC) linked via triazole groups was designed and synthesized. Its sensing behavior toward metal ions was investigated by ultraviolet-visible and fluorescence spectroscopies. Addition of a Fe(2+) ion to a tetrahydrofuran solution of HBTC gave a visual color change as well as a significantly quenched fluorescence emission, while other tested 19 metal ions induced no color or spectral changes. This compound was found to be highly selective and sensitive for Fe(2+) with a low limit of detection (2.03 μM) which is, to the best of our knowledge, the superior than the previously studied chemosensors for Fe(2+). Graphical Abstract ᅟ.

  9. A simple polyethylenimine-salicylaldehyde fluorescence probe: sensitive and selective detection of Zn2+ and Cd2+ in aqueous solution by adding S2- ion

    NASA Astrophysics Data System (ADS)

    Tan, Nai-Di; Yin, Jian-Hang; Pu, Guang; Yuan, Yaqing; Meng, Lei; Xu, Na

    2016-12-01

    A novel polyethylenimine (PEI)-salicylaldehyde (SA) fluorescence probe, simply prepared by stirring procedure, can sensitively detect Zn2+ (25 nM) and Cd2+ (10 nM) in aqueous solution. Formation of metal-to-ligand complexes (Zn2+@PEI-SA and Cd2+@PEI-SA) between ions and Schiff base generate a new emission at 454 nm, which increase with the concentration of the ions (turn-on). Furthermore, after adding S2- into the complexes, we can distinguish them due to the fluorescence quenching for Cd2+@PEI-SA (turn-off). Both response times for turning on and off the fluorescence are less than 1 min.

  10. Late Pleistocene granodiorite beneath Crater Lake caldera, Oregon, dated by ion microprobe

    USGS Publications Warehouse

    Bacon, C.R.; Persing, H.M.; Wooden, J.L.; Ireland, T.R.

    2000-01-01

    Variably melted granodiorite blocks ejected during the Holocene caldera-forming eruption of Mount Mazama were plucked from the walls of the climactic magma chamber ~15 km depth. Ion-microprobe U-Pb dating of zircons from two unmelted granodiorite blocks with SHRIMP RG (sensitive high-resolution ion microprobe-reverse geometry) gives a nominal 238U/206Pb age of 101+78-80 ka, or 174+89-115 ka when adjusted for an initial 230Th deficit. SHRIMP RG U-Th measurements on a subset of the zircons yield a 230Th/238U isochron age of 112 ?? 24 ka, considered to be the best estimate of the time of solidification of the pluton. These results suggest that the granodiorite is related to andesite and dacite of Mount Mazama and not to magmas of the climactic eruption. The unexposed granodiorite has an area of at least 28 km2. This young, shallow pluton was emplaced in virtually the same location where a similarly large magma body accumulated and powered violent explosive eruptions ~7700 yr ago, resulting in collapse of Crater Lake caldera.

  11. Determination of the topology of endoplasmic reticulum membrane proteins using redox-sensitive green-fluorescence protein fusions.

    PubMed

    Tsachaki, Maria; Birk, Julia; Egert, Aurélie; Odermatt, Alex

    2015-07-01

    Membrane proteins of the endoplasmic reticulum (ER) are involved in a wide array of essential cellular functions. Identification of the topology of membrane proteins can provide significant insight into their mechanisms of action and biological roles. This is particularly important for membrane enzymes, since their topology determines the subcellular site where a biochemical reaction takes place and the dependence on luminal or cytosolic co-factor pools and substrates. The methods currently available for the determination of topology of proteins are rather laborious and require post-lysis or post-fixation manipulation of cells. In this work, we have developed a simple method for defining intracellular localization and topology of ER membrane proteins in living cells, based on the fusion of the respective protein with redox-sensitive green-fluorescent protein (roGFP). We validated the method and demonstrated that roGFP fusion proteins constitute a reliable tool for the study of ER membrane protein topology, using as control microsomal 11β-hydroxysteroid dehydrogenase (11β-HSD) proteins whose topology has been resolved, and comparing with an independent approach. We then implemented this method to determine the membrane topology of six microsomal members of the 17β-hydroxysteroid dehydrogenase (17β-HSD) family. The results revealed a luminal orientation of the catalytic site for three enzymes, i.e. 17β-HSD6, 7 and 12. Knowledge of the intracellular location of the catalytic site of these enzymes will enable future studies on their biological functions and on the role of the luminal co-factor pool.

  12. High sensitivity detection of cancer in vivo using a dual-controlled activation fluorescent imaging probe based on H-dimer formation and pH activation.

    PubMed

    Ogawa, Mikako; Kosaka, Nobuyuki; Regino, Celeste A S; Mitsunaga, Makoto; Choyke, Peter L; Kobayashi, Hisataka

    2010-05-01

    The key to improving the sensitivity of in vivo molecular imaging is to increase the target-to-background signal ratio (TBR). Optical imaging has a distinct advantage over other molecular imaging methods in that the fluorescent signal can be activated at the target thus reducing background signal. Previously, we found that H-dimer formation quenches fluorescence of xanthene fluorophores, and among these, TAMRA had the highest quenching ratio. Another approach to lowering background signal is to employ pH activation based on the photon-induced electron transfer (PeT) theory. We hypothesized that combining these two strategies could lead to greater quenching capacity than was possible with either probe alone. A pH-sensitive fluorophore, pHrodo or TAMRA was conjugated to the cancer targeting molecules, avidin (Av) and trastuzumab (Tra). As expected, both pHrodo and TAMRA formed H-dimers when conjugated to avidin or antibody and the dimerization resulted in efficient fluorescence quenching. In addition, pHrodo conjugated probes showed pH-dependent fluorescence activation. When the probes were used in an in vivo animal model, fluorescence endoscopy with Av-pHrodo depicted tumors with high TBR 1 h and 2 h after injection. Av-TAMRA also visualized tumors 1 h and 2 h after the injection, however, TBR was lower due to the background signal from non-specific binding 1 h after the injection as well as background fluorescence from the unbound agent. Thus, we demonstrate that a dual-controlled activatable optical probe based on the combination of H-dimer formation and pH activation can achieve high TBR at early time points during in vivo molecular imaging.

  13. Depth Probing Soft X-ray Microprobe (DPSXRM) for High Resolution Probing of Earth's Microstructural Samples

    NASA Astrophysics Data System (ADS)

    Dikedi, P. N.

    2015-12-01

    The Cambrian explosion; occurrence of landslides in very dry weather conditions; rockslides; dead, shriveled-up and crumbled leaves possessing fossil records with the semblance of well preserved, flat leaves; abundance of trilobite tracks in lower and higher rock layers; and sailing stones are enigmas demanding demystifications. These enigmas could be elucidated when data on soil structure, texture and strength are provided by some device with submicrometre accuracy; for these and other reasons, the design of a Depth Probing Soft X-ray Microprobe (DPSXRM), is being proposed; it is expected to deliver soft X-rays, at spatial resolution, ϛ≥600nm and to probe at the depth of 0.5m in 17s. The microprobe is portable compared to a synchrotron radiation facility (Diamond Light Source has land size of 43,300m2); spatial resolution,ϛ , of the DPSXRM surpasses those of the X-ray Fluorescence microanalysis (10µm), electron microprobe (1-3µm) and ion microprobe (5->30µm); the DPSXRM has allowance for multiple targets. Vanadium and Manganese membranes are proposed owing to respective 4.952KeV VKα1 and 5.899KeV MnKα1 X-rays emitted, which best suits micro-probing of Earth's microstructural samples. Compound systems like the Kirk-Patrick and Baez and Wolter optics, aspheric mirrors like elliptical and parabolic optics, small apertures and Abbe sine condition are employed to reduce or remove astigmatism, obliquity, comatic and spherical aberrations—leading to good image quality. Results show that 5.899KeV MnKα1 and 4.952KeV VKα1 soft X-rays will travel a distance of 2.75mm to form circular patches of radii 2.2mm and 2.95mm respectively. Zone plate with nth zone radius of 1.5mm must be positioned 1.5mm and 2mm from the electron gun if circular patches must be formed from 4.952KeV VKα1 and 5.899KeV MnKα1 soft X-rays respectively. The focal lengths of 0.25μm≤ƒ≤1.50μm and 0.04μm≤ƒ≤0.2μm covered by 4.952KeV VKα1 and 5.899KeV Mn Kα1 soft X-Rays, will

  14. Radioactive halos and ion microprobe measurement of Pb isotope ratios

    NASA Technical Reports Server (NTRS)

    Gentry, R. V.

    1974-01-01

    This investigation was to obtain, if possible, the Pb isotope ratios of both lunar and meteoritic troilite grains by utilizing ion microprobe techniques. Such direct in situ measurement of Pb isotope ratios would eliminate contamination problems inherent in wet chemistry separation procedures, and conceivably determine whether lunar troilite grains were of meteoritic origin. For comparison purposes two samples of meteoritic troilite were selected (one from Canyon Diablo) for analysis along with two very small lunar troilite grains (approximately 50-100 microns). It was concluded that the ion microprobe as presently operating, does not permit the in situ measurement of Pb isotope ratios in lunar or meteoritic troilite. On the basis of these experiments no conclusions could be drawn as to the origin of the lunar troilite grains.

  15. A novel and sensitive fluorescence immunoassay for the detection of fluoroquinolones in animal-derived foods using upconversion nanoparticles as labels.

    PubMed

    Hu, Gaoshuang; Sheng, Wei; Zhang, Yan; Wu, Xuening; Wang, Shuo

    2015-11-01

    A novel fluorescence immunoassay to detect fluoroquinolones in animal-derived foods was developed for the first time by use of upconversion nanoparticles as signal-probe labels. The bioassay system was established by the use of coating-antigen-modified polystyrene particles as immune-sensing probes for separation and anti-norfloxacin monoclonal antibody conjugated with carboxyl-functionalized NaYF4:Yb,Er upconversion nanoparticles which were prepared via a pyrolysis method and a subsequent ligand exchange process as fluorescent-signal probes (emission intensity recorded at 542 nm with excitation at 980 nm). Under optimized conditions, detection of fluoroquinolones was performed easily. The detection limit of this fluorescence immunoassay for norfloxacin, for example, was 10 pg mL(-1), within a wide linear range of 10 pg mL(-1) to 10 ng mL(-1) (R (2)  = 0.9959). For specificity analysis, the data obtained indicate this method could be applied in broad-spectrum detection of fluoroquinolones. The recoveries of norfloxacin-spiked animal-derived foods ranged from 82.37 to 132.22 %, with coefficients of variation of 0.24-25.06 %. The extraction procedure was rapid and simple, especially for milk samples, which could be analyzed directly without any pretreatment. In addition, the results obtained with the method were in good agreement with those obtained with commercial ELISA kits. The fluorescence immunoassay was more sensitive, especially with regard to the detection limit in milk samples (0.01 ng mL(-1) for norfloxacin): it was 50-fold more sensitive than commercial ELISA kits (0.5 ng mL(-1) for norfloxacin). The results show the proposed fluorescence immunoassay was facile, sensitive, and interference free, and is an alternative method for the quantitative detection of fluoroquinolone residues in animal-derived foods.

  16. Microprobe analyses of glasses and minerals from Luna-16 soil

    NASA Technical Reports Server (NTRS)

    Brown, R. W.; Harmon, R. S.; Jakes, P.; Reid, A. M.; Ridley, W. I.; Warner, J. L.

    1971-01-01

    Electron microprobe analyses are presented for nine elements in 250 glasses and 434 pyroxenes, eight elements in 113 olivines, and six elements in 354 feldspars, 35 spinels, and 159 ilmenites. All grains are from the 125-425 micron fraction of horizon A and horizon D soil from the Luna 16 sample. A norm is presented for each glass analysis and the structural formula is calculated for each mineral analysis.

  17. 230Th-U dating of surficial deposits using the ion microprobe (SHRIMP-RG): A microstratigraphic perspective

    USGS Publications Warehouse

    Maher, K.; Wooden, J.L.; Paces, J.B.; Miller, D.M.

    2007-01-01

    We used the sensitive high-resolution ion microprobe reverse-geometry (SHRIMP-RG) to date pedogenic opal using the 230Th-U system. Due to the high-spatial resolution of an ion microprobe (typically 30 ??m), regions of pure opal within a sample can be targeted and detrital material can be avoided. In addition, because the technique is non-destructive, the sample can be preserved for other types of analyses including electron microprobe or other stable isotope or trace element ion microprobe measurements. The technique is limited to material with U concentrations greater than ???50 ppm. However, the high spatial resolution, small sample requirements, and the ability to avoid detrital material make this technique a suitable technique for dating many Pleistocene deposits formed in semi-arid environments. To determine the versatility of the method, samples from several different deposits were analyzed, including silica-rich pebble coatings from pedogenic carbonate horizons, a siliceous sinter deposit, and opaline silica deposited as a spring mound. U concentrations for 30-??m-diameter spots ranged from 50 to 1000 ppm in these types of materials. The 230Th/232Th activity ratios also ranged from ???100 to 106, eliminating the need for detrital Th corrections that reduce the precision of traditional U-Th ages for many milligram- and larger-sized samples. In pedogenic material, layers of high-U opal (ca. 500 ppm) are commonly juxtaposed next to layers of calcite with much lower U concentrations (1-2 ppm). If these types of samples are not analyzed using a technique with the appropriate spatial resolution, the ages may be strongly biased towards the age of the opal. Comparison with standard TIMS (Thermal Ionization Mass Spectrometry) measurements from separate microdrilled samples suggests that although the analytical precision of the ion microprobe (SHRIMP-RG) measurements is less than TIMS, the high spatial resolution results in better accuracy in the age determination for

  18. Spherical chamber effective solution for multipurpose nuclear microprobe

    NASA Astrophysics Data System (ADS)

    Pelicon, P.; Simčič, J.; Jakšić, M.; Medunić, Z.; Naab, F.; McDaniel, F. D.

    2005-04-01

    Vacuum chambers for multipurpose nuclear microprobes must provide for the installation and servicing of several detection systems operating simultaneously, as well as sample visual control and mechanical manipulation. Detectors for X-rays, scattered ions, nuclear reaction products, secondary electrons, secondary luminescence and optical microscopes are mounted at the angles preferably larger than 120° with respect to the beam direction. Their positioning should not increase the space in the region between the ion lens and the focal point of the microprobe. Spherical chambers presented here effectively solve this problem and offer, at the same time, ports for gamma-ray detector, annular microscope, easy manual access in the sample region, ports for vertical and horizontal sample positioning and manipulation, as well as STIM and ERDA detectors at forward scattering angles and the Faraday cup. The basic construction, resulting in the three different but similar chamber designs at three nuclear microprobes worldwide, are presented. Current installation details, comments on the performance and suggested improvements are given.

  19. A highly selective and sensitive fluorescence assay for determination of copper(II) and cobalt(II) ions in environmental water and toner samples.

    PubMed

    Tsai, Chia-Yi; Lin, Yang-Wei

    2013-02-21

    In this study, a highly selective and sensitive fluorescence assay has been proposed for the determination of copper(II) and cobalt(II) ions in environmental water and toner samples. In the presence of hydrogen peroxide, copper(II) reacted with a new fluorescence reagent Amplex® UltraRed (AUR), forming a fluorescence product only at pH 7.0, while the fluorescence product of cobalt(II) with AUR formed only at pH 9.0. The fluorescence signal obtained was linear with respect to the copper(II) concentration over the range of 1.6-12.0 μM (R(2) = 0.988) and was linear with respect to the cobalt(II) concentration over the range of 45.0 nM to 1.0 μM (R(2) = 0.992). The limits of detection (at a signal-to-noise ratio of 3) for copper(II) and cobalt(II) were 0.17 μM and 14.0 nM, respectively. Our present approach is simpler, faster, and more cost-effective than other techniques for the detection of copper(II) and cobalt(II) ions in environmental water samples and that of copper(II) ions in toner samples.

  20. A highly sensitive label-free sensor for Mercury ion (Hg²⁺) by inhibiting thioflavin T as DNA G-quadruplexes fluorescent inducer.

    PubMed

    Ge, Jia; Li, Xi-Ping; Jiang, Jian-Hui; Yu, Ru-Qin

    2014-05-01

    DNA sequences with guanine repeats can be induced to form G-quartets that adopt G-quadruplex structures in the presence of thioflavin T (ThT). ThT plays a dual role of inducing DNA sequences to fold into quadruplex structures and of sensing the change by its remarkable fluorescence enhancement. ThT binding to the DNA sequences with guanine repeats showed highly specific fluorescence enhancement compared with single/double-stranded DNA. In this work, we have utilized the conformational switch from G-quadruplex complex induced by fluorogenic dye ThT to Hg(2+) mediated T-Hg-T double-stranded DNA formation, thereby pioneering a facile approach to detect Hg(2+) with fluorescence spectrometry. Through this approach, Hg(2+) in aqueous solutions can be detected at 5 nM with fluorescence spectrometry in a facile way, with high selectivity against other metal ions. These results indicate the introduced label-free method for fluorescence spectrometric Hg(2+) detection is simple, quantitative, sensitive, and highly selective.

  1. Fluorescence enhancement of glutathione capped CdTe/ZnS quantum dots by embedding into cationic starch for sensitive detection of rifampicin

    NASA Astrophysics Data System (ADS)

    Hooshyar, Zari; Bardajee, Ghasem Rezanejade

    2017-02-01

    In this study, we describe the synthesis of a new quantum dots (QDs) by embedding glutathione capped CdTe/ZnS QDs into cationic starch biopolymer (CS-GSH-CdTe/ZnS QDs). The fluorescence intensity of prepared QDs was significantly enhanced. When QDs interacted with rifampicin, the fluorescence intensity of the CS-GSH-CdTe/ZnS QDs was highly quenched compared with GSH-CdTe/ZnS QDs. Based on the above, a new fluorescent nanosensor for simple, sensitive and selective detection of rifampicin was developed. The fluorescence quenching was well described by the typical Stern-Volmer equation. After optimization, the linear range of the as-prepared QDs fluorescence intensity versus the concentration of rifampicin was F0/F = 0.0422Q + 1.109 (R2 = 0.99). The detection limit was 0.06 × 10- 6 mol/L. The proposed method with satisfactory results was used to detect rifampicin in commercial capsules and tablets.

  2. A novel europium-sensitive fluorescent nano-chemosensor based on new functionalized magnetic core-shell Fe3O4@SiO2 nanoparticles.

    PubMed

    Ganjali, Mohammad Reza; Hosseini, Morteza; Khobi, Mehdi; Farahani, Shima; Shaban, Masoom; Faridbod, Farnoush; Shafiee, Abbas; Norouzi, Parviz

    2013-10-15

    A novel Eu(3+)-sensitive fluorescent chemosensor is introduced. It is based on magnetic core-shell silica nanoparticle which is functionalized by Cinchonidine (CD-Fe3O4@SiO2). The nano-chemosensor was synthesized and characterized by Fourier transform infrared spectroscopy (FT-IR), thermal gravimetric analysis (TGA), transmission electron microscopy (TEM), scanning electron microscopy (SEM), UV-visible absorption and fluorescence emission. The fluorescent nano-chemosensor shows a selective interaction with Eu(3+) ion. Fluorescence studies revealed that the emission intensity of the functionalized magnetic core-shell silica nanoparticles (CD-Fe3O4@SiO2 NPs) increases significantly by addition of various concentrations of Eu(3+) ion. While in case of mono, di, and other trivalent cations, weak changes or either no changes in intensity were observed. The enhancement in fluorescence intensity of nano-chemosensor is because of the strong covalent binding of Eu(3+) ion to CD-Fe3O4@SiO2 NPs with a large binding constant value of 1.7 × 10(5) mol L(-1).

  3. Staurosporine as an agonist for induction of GLUT4 translocation, identified by a pH-sensitive fluorescent IRAP-mOrange2 probe.

    PubMed

    Li, Yufeng; Zheng, Li; Wang, Dan; Zhang, Xiang; Li, Jia; Ali, Sher; Lu, Jingze; Zong, Hao; Xu, Xiaolan

    2016-11-25

    Insulin-stimulated GLUT4 translocation from GLUT4 storage vesicles (GSVs) to the plasma membrane (PM) constitutes a key process for blood glucose control. Therefore, compounds that could promote GLUT4 translocation into the PM represent potential drugs for the treatment of diabetes. In this research, we screened for agonists that induce GLUT4 translocation by using a novel pH-sensitive fluorescent probe, insulin-regulated aminopeptidase (IRAP)-mOrange2. We identified as well as validated one agonist, staurosporine, from a 64,000 compound library. Staurosporine promotes GSVs translocation into the PM and increases glucose uptake through the AMP-activated protein kinase (AMPK) pathway, serving as an effective insulin additive analogue in L6 cells. Our work highlights the convenience and efficiency of this novel pH-sensitive fluorescent probe and reveals the new biological activity of staurosporine as an agonist for GLUT4 translocation and as an effective insulin additive analogue.

  4. A Sensitive and Label-Free Pb(II) Fluorescence Sensor Based on a DNAzyme Controlled G-Quadruplex/Thioflavin T Conformation

    PubMed Central

    Wen, Yanli; Wang, Lele; Li, Lanying; Xu, Li; Liu, Gang

    2016-01-01

    Pb(II) can cause serious damaging effects to human health, and thus, the study of Pb2+ detection methods to sensitively and selectively monitor Pb(II) pollution has significant importance. In this work, we have developed a label-free fluorescence sensing strategy based on a Pb(II) DNAzyme cleavage and the ThT/G-quadruplex complex. In the presence of Pb(II), a G-rich tail was cut and released from the substrate strand, which then would form a G-quadruplex structure by combination with ThT dye. The fluorescence signal increase was then measured for sensitive Pb(II) quantification with a limit of detection of 0.06 nM. Our sensor also demonstrated high selectivity against six different metal ions, which is very important for the analysis of complex samples. PMID:27999248

  5. Sensitive and background-free determination of thiols from wastewater samples by MOF-5 extraction coupled with high-performance liquid chromatography with fluorescence detection using a novel fluorescence probe of carbazole-9-ethyl-2-maleimide.

    PubMed

    Lv, Zhengxian; Sun, Zhiwei; Song, Cuihua; Lu, Shuaimin; Chen, Guang; You, Jinmao

    2016-12-01

    A sensitive and background-free pre-column derivatization method for the determination of thiol compounds using metal-organic framework material (MOF-5) as dispersive solid-phase extraction (DSPE) adsorbent followed by high-performance liquid chromatography fluorescence detection (HPLC-FLD) has been developed. In this paper, a novel labeling reagent, carbazole-9-ethyl-2-maleimide(CAEM), was synthesized and reacted with thiols at 40°C for 10min in the presence of PBS buffer (0.02mol/L, pH 7.5). Interestingly, CAEM itself had no fluorescence, while its derivatives exhibited intense fluorescence with an excitation maximum at λex 274nm and an emission maximum at λem 363nm, which greatly reduced the background interference and improved the sensitivity of the method. Furthermore, the MOF-5 was prepared and used as DSPE adsorbent for the selective adsorption of thiols from wastewater sample. Under the optimized experimental conditions, an excellent linearity for all analytes over their concentration ranges of 0.01-1.0μmol/L (R(2)>0.9986)were obtained with the limit of detection (LOD) ranging from 8 to 17.1pmol/L for nine tested thiols. The feasibility of this method for the determination of thiols in wastewater samples had been evaluated and satisfactory average recoveries (n=3) were achieved with the range of 86.6-98.5%.

  6. Correlated petrographic, electron microprobe, and ion microprobe studies of selected primitive and processed phase assemblages in meteorites

    NASA Technical Reports Server (NTRS)

    Albee, Arden L.

    1993-01-01

    During the past three years we have received support to continue our research in elucidating the formation and alteration histories of selected meteoritic materials by a combination of petrographic, trace element, and isotopic analyses employing optical and scanning electron microscopes and electron and ion microprobes. The awarded research funds enabled the P.I. to attend the annual LPSC, the co-I to devote approximately 15 percent of his time to the research proposed in the grant, and partial support for a visiting summer post-doctoral fellow to conduct electron microprobe analyses of meteoritic samples in our laboratory. The research funds, along with support from the NASA Education Initiative awarded to P.I. G. Wasserburg, enabled the co-I to continue a mentoring program with inner-city minority youth. The support enabled us to achieve significant results in the five projects that we proposed (in addition to the Education Initiative), namely: studies of the accretional and post-accretional alteration and thermal histories in CV meteorites, characterization of periclase-bearing Fremdlinge in CV meteorites, characterization of Ni-Pt-Ge-Te-rich Fremdlinge in CV meteorites in an attempt to determine the constraints they place on the petrogenetic and thermal histories of their host CAI's, correlated electron and ion microprobe studies of silicate and phosphate inclusions in the Colomera meteorite in an attempt to determine the petrogenesis of the IE iron meteorites, and development of improved instrumental and correction procedures for improved accuracy of analysis of meteoritic materials with the electron microprobe. This grant supported, in part or whole, 18 publications so far by our research team, with at least three more papers anticipated. The list of these publications is included. The details of the research results are briefly summarized.

  7. Photoactivation and calcium sensitivity of the fluorescent NO indicator 4,5-diaminofluorescein (DAF-2): implications for cellular NO imaging.

    PubMed

    Broillet, M; Randin, O; Chatton, J

    2001-03-02

    The fluorescent indicator of nitric oxide (NO), 4,5-diaminofluorescein (DAF-2), and its membrane-permeable derivative (DAF-2 diacetate) have been recently developed to perform real-time biological imaging of NO. In this study, we show that DAF-2 is strongly influenced by factors other than the concentration of NO itself. Using measurements with a fluorimeter as well as fluorescence microscopy, we found that the divalent cation concentration in the medium, as well as the incident light, strongly affects the ability of DAF-2 to detect NO. Calcium, in particular, enhanced the signal detection of NO released by NO donors by up to 200 times. With multiple and longer exposures to light, no bleaching of the dye was observed but, instead, a potentiation of the fluorescence response could be measured. While these two properties will affect the use and interpretation of the hitherto acquired data with this fluorescent compound, they may also open up new possibilities for its application.

  8. Simple and sensitive synchronous- fluorescence method for the determination of trace bisphenol S based on its inhibitory effect on the fluorescence quenching reaction of rhodamine B.

    PubMed

    Cao, Gui-ping; Chen, Ting; Zhuang, Ya-feng

    2013-07-01

    An inhibitory kinetic fluorimetric method is reported for the determination of trace bisphenol S (BPS). The proposed method is based on the inhibitory effect of BPS on the fluorescence quenching of rhodamine B (RhB) caused by potassium bromate in a dilute phosphoric acid medium. Under the optimal conditions of the experiment, the detection limit for BPS was 0.021 mg/L, and the linear range of determination was from 0.035 mg/L to 0.750 mg/L. The relative standard deviations of 11 measurements for 0.20 mg/L and 0.40 mg/L BPS solutions were 2.74 % and 1.87 %, respectively. The method was successfully applied to the determination of bisphenol S derived from commercially available plastic film samples in hot water. A possible reaction mechanism of the inhibitory effect of BPS on the fluorescence quenching of RhB was proposed.

  9. An X-Ray Microprobe for In-Situ Stone and Wood Characterization

    NASA Astrophysics Data System (ADS)

    Lovoi, P.; Asmus, J. F.

    NonDestructive Testing (NDT) has become an essential ingredient in the conservation of artworks and in the preservation of historic buildings. In many instances it is necessary to characterize the underlying strata of an artistic or historic object in order to plan technical conservation measures, to understand its history, to authenticate it, or to search for hidden features. X-ray and gamma-ray radiography as well as infrared imaging have been ubiquitous in conservation practice for generations. Recent decades have also seen the introduction of ultrasonic imaging, thermovision, x-ray fluorescence, neutron activation analyses, holographic interferometry, isotopic and trace element analyses, the electron microprobe, the laser microprobe, microwave impulse radar, eddy current imaging, and fiber-optic imaging. Unfortunately, for mainstream conservation and preservation some of these technologies are too costly or difficult to be implemented in any general way. In other instances penetration is too superficial or signals from the depth of interest are masked by interferences. Nevertheless, sufficiently important problems have arisen to warrant the utilization of each of the above NDT technologies as well as still others. A new diagnostic device has been introduced into the conservation field. Stone characterization analyses are reported using miniature x-ray devices that can be inserted into cracks and holes in specimens of interest. The family of x-ray tubes employed in these studies range in diameter from 1 to 6 mm. Operating voltages up to 50 kV are available. Electrical power and cooling are delivered through a flexible cable that has a bend diameter of less than 3 cm. Thus, it was possible to insert the x-ray tube into small holes and cracks in marble stones. In this manner radiographs of the outer strata of stones (and embedded metal pins) have been produced without having to transmit through the entire thickness of large blocks. It should also be possible to

  10. A reduced graphene oxide-based fluorescence resonance energy transfer sensor for highly sensitive detection of matrix metalloproteinase 2.

    PubMed

    Xi, Gaina; Wang, Xiaoping; Chen, Tongsheng

    2016-01-01

    A novel fluorescence nanoprobe (reduced nano-graphene oxide [nrGO]/fluorescein isothiocyanate-labeled peptide [Pep-FITC]) for ultrasensitive detection of matrix metalloproteinase 2 (MMP2) has been developed by engineering the Pep-FITC comprising the specific MMP2 substrate domain (PLGVR) onto the surface of nrGO particles through non-covalent linkage. The nrGO was obtained by water bathing nano-graphene oxide under 90°C for 4 hours. After mixing the nrGO and Pep-FITC for 30 seconds, the fluorescence from Pep-FITC was almost completely quenched due to the fluorescence resonance energy transfer between fluorescein isothiocyanate (FITC) and nrGO. Upon cleavage of the amide bond between Leu and Gly in the Pep-FITC by protease-MMP2, the FITC bound to nrGO was separated from nrGO surface, disrupting the fluorescence resonance energy transfer process and resulting in fluorescence recovery of FITC. Under optimal conditions, the fluorescence recovery of nrGO/Pep-FITC was found to be directly proportional to the concentration of MMP2 within 0.02-0.1 nM. The detection limit of the nrGO/Pep-FITC was determined to be 3 pM, which is approximately tenfold lower than that of the unreduced carboxylated nano-graphene oxide/Pep-FITC probe.

  11. A reduced graphene oxide-based fluorescence resonance energy transfer sensor for highly sensitive detection of matrix metalloproteinase 2

    PubMed Central

    Xi, Gaina; Wang, Xiaoping; Chen, Tongsheng

    2016-01-01

    A novel fluorescence nanoprobe (reduced nano-graphene oxide [nrGO]/fluorescein isothiocyanate-labeled peptide [Pep-FITC]) for ultrasensitive detection of matrix metalloproteinase 2 (MMP2) has been developed by engineering the Pep-FITC comprising the specific MMP2 substrate domain (PLGVR) onto the surface of nrGO particles through non-covalent linkage. The nrGO was obtained by water bathing nano-graphene oxide under 90°C for 4 hours. After mixing the nrGO and Pep-FITC for 30 seconds, the fluorescence from Pep-FITC was almost completely quenched due to the fluorescence resonance energy transfer between fluorescein isothiocyanate (FITC) and nrGO. Upon cleavage of the amide bond between Leu and Gly in the Pep-FITC by protease-MMP2, the FITC bound to nrGO was separated from nrGO surface, disrupting the fluorescence resonance energy transfer process and resulting in fluorescence recovery of FITC. Under optimal conditions, the fluorescence recovery of nrGO/Pep-FITC was found to be directly proportional to the concentration of MMP2 within 0.02–0.1 nM. The detection limit of the nrGO/Pep-FITC was determined to be 3 pM, which is approximately tenfold lower than that of the unreduced carboxylated nano-graphene oxide/Pep-FITC probe. PMID:27143876

  12. A sensitive dual-fluorescence reporter system enables positive selection of ras suppressors by suppression of ras-induced apoptosis.

    PubMed

    Dolnikov, Alla; Shen, Sylvie; Millington, Michelle; Passioura, Toby; Pedler, Michelle; Rasko, John Edward Joshua; Symonds, Geoff

    2003-10-01

    We have developed a novel dual-fluorescence reporter system incorporating green (GFP) and red (RFP) fluorescent proteins to monitor expression of the N-ras(m) gene and an N-ras(m) suppressor, respectively. Retroviral vectors were produced in which human N-ras(m) (codon 13 mutation) was coexpressed with GFP, and a ribozyme specifically targeting N-ras(m) was coexpressed with RFP. N-Ras(m) suppression was monitored by measurement of GFP fluorescence in dual-fluorescent (GFP and RFP) cells. We demonstrated that the degree of N-ras(m) suppression was dependent on the ribozyme dose, proportional to red fluorescence, in dual-fluorescent cells. We further showed that ribozyme-mediated N-ras(m)suppression inhibited growth of NIH3T3 and CD34-positive TF-1 cells. In these cultures, ras suppressor activity resulted in the depletion of suppressor-positive cells due to inhibition of cell growth. In contrast, N-ras(m) suppression produced a growth advantage to human leukemic K562 cells, presumably by inhibiting N-ras(m)-induced apoptosis. In K562 cells, ras suppression resulted in the outgrowth of suppressor-positive cells. This provides a platform to identify suppressors of ras that is based on function.

  13. Enzyme-free and label-free fluorescence aptasensing strategy for highly sensitive detection of protein based on target-triggered hybridization chain reaction amplification.

    PubMed

    Wang, Xiuzhong; Jiang, Aiwen; Hou, Ting; Li, Haiyin; Li, Feng

    2015-08-15

    Proteins are of great importance in medical and biological fields. In this paper, a novel fluorescent aptasensing strategy for protein assay has been developed based on target-triggered hybridization chain reaction (HCR) and graphene oxide (GO)-based selective fluorescence quenching. Three DNA probes, a helper DNA probe (HP), hairpin probe 1 (H1) and hairpin probe 2 (H2) are ingeniously designed. In the presence of the target, the aptamer sequences in HP recognize the target to form a target-aptamer complex, which causes the HP conformation change, and then triggers the chain-like assembly of H1 and H2 through the hybridization chain reaction, generating a long chain of HP leading complex of H1 and H2. At last the fluorescence indicator SYBR Green I (SG) binds with the long double strands of the HCR product through both intercalation and minor groove binding. When GO was added into the solutions after HCR, the free H1, H2 and SG would be closely adsorbed onto GO surface via π-π stacking. However, the HCR product cannot be adsorbed on GO surface, thereby the SG bound to HCR product gives a strong fluorescence signal dependent on the concentration of the target. With the use of platelet-derived growth factor BB (PDGF-BB) as the model analyte, this newly designed protocol provides a highly sensitive fluorescence detection of PDGF-BB with a limit of detection down to 1.25 pM, and also exhibit good selectivity and applicability in complex matrixes. Therefore, the proposed aptasensing strategy based on target-triggered hybridization chain reaction amplification should have wide applications in the diagnosis of genetic diseases due to its simplicity, low cost, and high sensitivity at extremely low target concentrations.

  14. Paper-based fluorescent sensor for rapid naked-eye detection of acetylcholinesterase activity and organophosphorus pesticides with high sensitivity and selectivity.

    PubMed

    Chang, Jiafu; Li, Haiyin; Hou, Ting; Li, Feng

    2016-12-15

    Various strategies have been proposed for the sensing of acetylcholinesterase (AChE) activity and organophosphorus pesticides (OPs). However, the practical application of most methods is restricted by their intrinsic drawbacks such as complexity, long analysis time, and high cost. Thus, it is highly desirable to develop simple, fast and sensitive approaches for AChE activity and OPs detection. Herein, we reported a simple paper-based fluorescent sensor (PFS) based on the aggregation induced emission (AIE) effect of tetraphenylethylene (TPE) and the addition reaction capability of maleimide, which has been used as a powerful tool for rapid naked-eye detection of AChE activity and OPs. The introduction of TPE provides the probe with unique fluorescence property in solid state and is of great importance for improving the sensitivity of PFS. The hydrolysis product of acetylthiocholine catalyzed by AChE induced the maleimide ring destruction and activated the fluorescence performance of TPE. Given that AChE activity can be specifically inhibited by OPs, the as-proposed PFS can also be utilized for sensitive detection of OPs. Meanwhile, the variation of fluorescence signal can be readily detected by naked eyes, and low detection limits of 2.5mUmL(-1) and 0.5ngmL(-1) for AChE activity and OPs are obtained, respectively. Moreover, it has been successfully applied for AChE activity and OPs detection in diluted human serum samples, showing its great potential to be applied in real samples. Thus, this strategy possesses considerable advantages of simplicity, rapid detection, portability, cost efficiency and visualization.

  15. Magic sized ZnS quantum dots as a highly sensitive and selective fluorescence sensor probe for Ag+ ions.

    PubMed

    Mandal, Abhijit; Dandapat, Anirban; De, Goutam

    2012-02-07

    A green and simple chemical synthesis of magic sized water soluble blue-emitting ZnS quantum dots (QDs) has been accomplished by reacting anhydrous Zn acetate, sodium sulfide and thiolactic acid (TLA) at room temperature in aqueous solution. Refluxing of this mixture in open air yielded ZnS clusters of about 3.5 nm in diameter showing very strong and narrow photoluminescence properties with long stability. Refluxing did not cause any noticeable size increment of the clusters. As a result, the QDs obtained after different refluxing conditions showed similar absorption and photoluminescence (PL) features. Use of TLA as a capping agent effectively yielded such stable and magic sized QDs. The as-synthesized and 0.5 h refluxed ZnS QDs were used as a fluorescence sensor for Ag(+) ions. It has been observed that after addition of Ag(+) ions of concentration 0.5-1 μM the strong fluorescence of ZnS QDs was almost quenched. The quenched fluorescence can be recovered by adding ethylenediamine to form a complex with Ag(+) ions. The other metal ions (K(+), Ca(2+), Au(3+), Cu(2+), Fe(3+), Mn(2+), Mg(2+), Co(2+)) showed little or no effect on the fluorescence of ZnS QDs when tested individually or as a mixture. In the presence of all these ions, Ag(+) responded well and therefore ZnS QDs reported in this work can be used as a Ag(+) ion fluorescence sensor.

  16. A sensitive fluorescent nanosensor for chloramphenicol based on molecularly imprinted polymer-capped CdTe quantum dots.

    PubMed

    Amjadi, Mohammad; Jalili, Roghayeh; Manzoori, Jamshid L

    2016-05-01

    A novel fluorescent nanosensor using molecularly imprinted silica nanospheres embedded CdTe quantum dots (CdTe@SiO2 @MIP) was developed for detection and quantification of chloramphenicol (CAP). The imprinted sensor was prepared by synthesis of molecularly imprinting polymer (MIP) on the hydrophilic CdTe quantum dots via reverse microemulsion method using small amounts of solvents. The resulting CdTe@SiO2 @MIP nanoparticles were characterized by fluorescence, UV-vis absorption and FT-IR spectroscopy and transmission electron microscopy. They preserved 48% of fluorescence quantum yield of the parent quantum dots. CAP remarkably quenched the fluorescence of prepared CdTe@SiO2 @MIP, probably via electron transfer mechanism. Under the optimal conditions, the relative fluorescence intensity of CdTe@SiO2 @MIP decreased with increasing CAP by a Stern-Volmer type equation in the concentration range of 40-500 µg L(-1). The corresponding detection limit was 5.0 µg L(-1). The intra-day and inter-day values for the precision of the proposed method were all <4%. The developed sensor had a good selectivity and was applied to determine CAP in spiked human and bovine serum and milk samples with satisfactory results.

  17. X-ray Mapping of Terrestrial and Extraterrestrial Materials Using the Electron Microprobe

    NASA Technical Reports Server (NTRS)

    Carpenter, P.

    2006-01-01

    the sample at microscopic and macroscopic scales with relatively high sensitivity, (2) Determine the modal abundance of minerals, and (3) Identify and relocate discrete features of interest in terms of size and chemistry. The coupled substitution of cations in minerals can result in significant variation in mineral chemistry, but at similar average Z, leading to poor backscattered-electron (BSE) contrast discrimination of mineralogy. It is necessary to discriminate phase chemistry at both the trace element level and the major element level. To date, the WDS of microprobe systems is preferred for mapping due to high throughput and the ability to obtain the necessary intensity to discriminate phases at both trace and major element concentrations. It is desirable to produce fully quantitative compositional maps of geological materials, which requires the acquisition of k-ratio maps that are background and dead-time corrected, and which have been corrected by phi(delta z> or an equivalent algorithm at each pixel. To date, turnkey systems do not allow the acquisition of k-ratio maps and the rigorous correction in this manner. X-ray maps of a chondrule from the Ourique meteorite, and a comb-layered xenolith from the San Francisco volcanic field, have been analyzed and processed to extract phase information. The Ourique meteorite presents a challenge due to relatively low BSE contrast, and has been studied using spectrum imaging. X-ray maps for Si, Mg, and FeK(alpha) were used to produce RGB images. The xenolith sample contains sector-zoned augite, olivine, plagioclase, and basaltic glass. X-ray maps were processed using Lispix and ImageJ software to produce mineral phase maps. The x-ray maps for Mg, Ca, and Ti were used with traceback to generate binary images that were converted to RGB images. These approaches are successful in discriminating phases, but it is desirable to achieve the methods that were used on lunar samples 30 years ago on current microprobe systems. Curnt

  18. Sensitive fluorescence detection of ATP based on host-guest recognition between near-infrared β-Cyclodextrin-CuInS2 QDs and aptamer.

    PubMed

    Hu, Tianyu; Na, Weidan; Yan, Xu; Su, Xingguang

    2017-04-01

    We have developed a near-infrared (NIR) fluorescent aptamer-based sensor for sensitive detection of adenosine-5'-triphosphate (ATP) by using a ATP-binding aptamer and β-Cyclodextrin-CuInS2 quantum dots (β-CD-CuInS2 QDs). The fluorescence of β-CD-CuInS2 QDs has a slight enhancement with the addition of ATP-binding aptamer due to the host-guest recognition between aptamer and β-CD. When ATP is added, it will bind to aptamer to form G-quadruplexes. Aptamer-ATP complexes can enter into the hydrophobic cavities of β-CD and result in great enhancement of the fluorescence intensity. Under the optimum conditions, the fluorescence intensity of β-CD-CuInS2 QDs is proportional to the concentration of ATP, which shows a good linear response toward ATP concentration range of 6-1200μmolL(-1), the detection limit for ATP is 3μmolL(-1). The present assay shows a good selectivity for ATP over other biologically important proteins, and it is applied to the determination of ATP in human serum sample with satisfactory results.

  19. Conformation-sensitive nucleoside analogues as topology-specific fluorescence turn-on probes for DNA and RNA G-quadruplexes

    PubMed Central

    Tanpure, Arun A.; Srivatsan, Seergazhi G.

    2015-01-01

    Development of probes that can discriminate G-quadruplex (GQ) structures and indentify efficient GQ binders on the basis of topology and nucleic acid type is highly desired to advance GQ-directed therapeutic strategies. In this context, we describe the development of minimally perturbing and environment-sensitive pyrimidine nucleoside analogues, based on a 5-(benzofuran-2-yl)uracil core, as topology-specific fluorescence turn-on probes for human telomeric DNA and RNA GQs. The pyrimidine residues of one of the loop regions (TTA) of telomeric DNA and RNA GQ oligonucleotide (ON) sequences were replaced with 5-benzofuran-modified 2′-deoxyuridine and uridine analogues. Depending on the position of modification the fluorescent nucleoside analogues distinguish antiparallel, mixed parallel-antiparallel and parallel stranded DNA and RNA GQ topologies from corresponding duplexes with significant enhancement in fluorescence intensity and quantum yield. Further, these GQ sensors enabled the development of a simple fluorescence binding assay to quantify topology- and nucleic acid-specific binding of small molecule ligands to GQ structures. Together, our results demonstrate that these nucleoside analogues are useful GQ probes, which are anticipated to provide new opportunities to study and discover efficient G-quadruplex binders of therapeutic potential. PMID:26202965

  20. Microwave assisted one-pot synthesis of graphene quantum dots as highly sensitive fluorescent probes for detection of iron ions and pH value.

    PubMed

    Zhang, Chunfang; Cui, Yanyan; Song, Li; Liu, Xiangfeng; Hu, Zhongbo

    2016-04-01

    Recently, carbon nanomaterials have received considerable attention as fluorescent probes owing to their low toxicity, water solubility and stable photochemical properties. However, the development of graphene quantum dots (GQDs) is still on its early stage. In this work, GQDs were successfully synthesized by one-step microwave assisted pyrolysis of aspartic acid (Asp) and NH4HCO3 mixture. The as-prepared GQDs exhibited strongly blue fluorescence with high quantum yield up to 14%. Strong fluorescence quenching effect of Fe(3+) on GQDs can be used for its high selectivity detection among of general metal ions. The probe exhibited a wide linear response concentration range (0-50 μM) to Fe(3+) and the limit of detection (LOD) was calculated to be 0.26 μM. In addition, GQDs are also sensitive to the pH value in the range from 2 to 12 indicating a great potential as optical pH sensors. More importantly, the GQDs possess lower cellular toxicity and high photostability and can be directly used as fluorescent probes for cell imaging.

  1. Development and utilization of a fluorescence-based receptor-binding assay for the site 5 voltage-sensitive sodium channel ligands brevetoxin and ciguatoxin.

    PubMed

    McCall, Jennifer R; Jacocks, Henry M; Niven, Susan C; Poli, Mark A; Baden, Daniel G; Bourdelais, Andrea J

    2014-01-01

    Brevetoxins are a family of ladder-frame polyether toxins produced during blooms of the marine dinoflagellate Karenia brevis. Consumption of fish exposed to K. brevis blooms can lead to the development of neurotoxic shellfish poisoning. The toxic effects of brevetoxins are due to activation of voltage-sensitive sodium channels (VSSCs) in cell membranes. Binding of toxins has historically been measured using a radioligand competition assay that is fraught with difficulty. In this study, we developed a novel fluorescence-based binding assay for the brevetoxin receptor. Several fluorophores were conjugated to polyether brevetoxin-2 and used as the labeled ligand. Brevetoxin analogs were able to compete for binding with the fluorescent ligands. This assay was qualified against the standard radioligand receptor assay for the brevetoxin receptor. Furthermore, the fluorescence-based assay was used to determine relative concentrations of toxins in raw extracts of K. brevis culture, and to determine ciguatoxin affinity to site 5 of VSSCs. The fluorescence-based assay was quicker, safer, and far less expensive. As such, this assay can be used to replace the current radioligand assay and will be a vital tool for future experiments examining the binding affinity of various ligands for site 5 on sodium channels.

  2. High-Resolution Ultrasound-Switchable Fluorescence Imaging in Centimeter-Deep Tissue Phantoms with High Signal-To-Noise Ratio and High Sensitivity via Novel Contrast Agents

    PubMed Central

    Cheng, Bingbing; Bandi, Venugopal; Wei, Ming-Yuan; Pei, Yanbo; D’Souza, Francis; Nguyen, Kytai T.; Hong, Yi; Yuan, Baohong

    2016-01-01

    For many years, investigators have sought after high-resolution fluorescence imaging in centimeter-deep tissue because many interesting in vivo phenomena—such as the presence of immune system cells, tumor angiogenesis, and metastasis—may be located deep in tissue. Previously, we developed a new imaging technique to achieve high spatial resolution in sub-centimeter deep tissue phantoms named continuous-wave ultrasound-switchable fluorescence (CW-USF). The principle is to use a focused ultrasound wave to externally and locally switch on and off the fluorophore emission from a small volume (close to ultrasound focal volume). By making improvements in three aspects of this technique: excellent near-infrared USF contrast agents, a sensitive frequency-domain USF imaging system, and an effective signal processing algorithm, for the first time this study has achieved high spatial resolution (~ 900 μm) in 3-centimeter-deep tissue phantoms with high signal-to-noise ratio (SNR) and high sensitivity (3.4 picomoles of fluorophore in a volume of 68 nanoliters can be detected). We have achieved these results in both tissue-mimic phantoms and porcine muscle tissues. We have also demonstrated multi-color USF to image and distinguish two fluorophores with different wavelengths, which might be very useful for simultaneously imaging of multiple targets and observing their interactions in the future. This work has opened the door for future studies of high-resolution centimeter-deep tissue fluorescence imaging. PMID:27829050

  3. Rapid and Sensitive Detection of Protein Biomarker Using a Portable Fluorescence Biosensor based on Quantum Dots and a Lateral Flow Test Strip

    SciTech Connect

    Li, Zhaohui; Wang, Ying; Wang, Jun; Tang, Zhiwen; Pounds, Joel G.; Lin, Yuehe

    2010-08-15

    A portable fluorescence biosensor with rapid and ultrasensitive response for trace protein has been built up with quantum dots and lateral flow test strip. The superior signal brightness and high photostability of quantum dots are combined with the promising advantages of lateral flow test strip and resulted in high sensitivity, selectivity and speedy for protein detection. Nitrated ceruloplasmin, a significant biomarker for cardiovascular disease, lung cancer and stress response to smoking, was used as model protein to demonstrate the good performances of this proposed Qdot-based lateral flow test strip. Quantitative detection of nitrated ceruloplasmin was realized by recording the fluorescence intensity of quantum dots captured on the test line. Under optimal conditions, this portable fluorescence biosensor displays rapid responses for nitrated ceruloplasmin in wide dynamic range with a detection limit of 0.1ng/mL (S/N=3). Furthermore, the biosensor was successfully utilized for spiked human plasma sample detection with the concentration as low as 1ng/mL. The results demonstrate that the quantum dot-based lateral flow test strip is capable for rapid, sensitive, and quantitative detection of nitrated ceruloplasmin and hold a great promise for point-of-care and in field analysis of other protein biomarkers.

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

    PubMed

    Yang, Qiong; Tan, Xuanping; Yang, Jidong

    2016-02-01

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

  5. Upconversion fluorescence and its thermometric sensitivity of Er3+:Yb3+ co-doped SrF2 powders prepared by combustion synthesis

    NASA Astrophysics Data System (ADS)

    Rakov, Nikifor; Maciel, Glauco S.; Xiao, Mufei

    2014-09-01

    Upconversion fluorescence of co-doped Er3+:Yb3+:SrF2 powders prepared by combustion synthesis was investigated under near-infrared ( λ = 980 nm) continuous wave laser excitation. Surface morphology of the samples and structures of the Er3+:Yb3+:SrF2 powders were studied with scanning electronic microscopy, energy dispersive x-ray, and x-ray powder diffraction. The spectrum of the fluorescence contains bands centered at ~410, ~522, ~545 and ~660 nm, corresponding respectively to transitions from upper levels 2H9/2, 2H11/2, 4S3/2 and 4F9/2 to the ground state 4I15/2, which can be identified as 4 f-4 f transitions from Er3+ excited states. In addition, the fluorescence is found sensitive to the temperature, which suggests that an optical temperature sensor would be feasible. The maximum sensitivity of the proposed sensor was found 0.00396 K-1.

  6. Surface Molecular Imprinting on Silica-Coated CdTe Quantum Dots for Selective and Sensitive Fluorescence Detection of p-aminophenol in Water.

    PubMed

    Lu, Xialin; Wei, Fangdi; Xu, Guanhong; Wu, Yanzi; Yang, Jing; Hu, Qin

    2017-01-01

    In this paper, a selective and sensitive sensor for the determination of p-aminophenol (PAP) was developed by grafting molecularly imprinted polymers (MIPs) on the surface of silica-coated CdTe quantum dots (CdTe@SiO2@MIPs). The obtained CdTe@SiO2@MIPs were characterized by X-ray powder diffraction, Fourier transform infrared spectroscopy, transmission electron microscopy and fluorescence spectroscopy. The fluorescence intensity of CdTe@SiO2@MIPs was more strongly quenched by PAP than that of the structural analogues of PAP. Under the optimal conditions, the fluorescence intensity of the CdTe@SiO2@MIPs decreased sensitively with the increase of PAP concentration in the range of 0.05-50 μM. The limit of detection was 0.02 μM (3σ/K sv). The sensor was successfully used to determine PAP in tap and lake water samples, and the average recoveries of PAP at various spiking levels ranged from 97.33 % to 103.3 % with relative standard deviations below 20 %.

  7. Layer-by-layer engineering fluorescent polyelectrolyte coated mesoporous silica nanoparticles as pH-sensitive nanocarriers for controlled release

    NASA Astrophysics Data System (ADS)

    Du, Pengcheng; Zhao, Xubo; Zeng, Jin; Guo, Jinshan; Liu, Peng

    2015-08-01

    Fluorescent core/shell composite has been fabricated by the layer-by-layer (LbL) assembly of the fluorescein isothiocyanate modified chitosan (CS-FITC) and sodium alginate (AL) onto the carboxyl modified mesoporous silica nanoparticles (MSN-COOH), followed by PEGylation. It exhibits stability in high salt-concentration media and the pH responsive fluorescent feature can be used for cell imaging. Furthermore, the modified MSN cores can enhance the DOX loading capacity and the multifunctional polyelectrolyte shell can adjust the drug release upon the media pH, showing a low leakage quantity at the neutral environment but significantly enhanced release at lower pH media mimicking the tumor environments. Therefore, the biocompatible fluorescent polyelectrolyte coated mesoporous silica nanoparticles (MSN-LBL-PEG) offer promise for tumor therapy.

  8. Proton beam micromachined resolution standards for nuclear microprobes

    NASA Astrophysics Data System (ADS)

    Watt, F.; Rajta, I.; van Kan, J. A.; Bettiol, A. A.; Osipowicz, T.

    2002-05-01

    The quest for smaller spot sizes has long been the goal of many nuclear microprobe groups worldwide, and consequently there is a need for good quality resolution standards. Such standards have to be consistent with the accurate measurement of state-of-the-art nuclear microbeam spot sizes, i.e. 400 nm for high current applications such as Rutherford backscattering spectrometry and proton-induced X-ray emission, and 100 nm for low current applications such as scanning transmission ion microscopy or ion beam-induced charge. The criteria for constructing a good quality nuclear microprobe resolution standard is therefore demanding: the standard has to be three dimensional with a smooth surface, have an edge definition better than the state-of-the-art beam spot resolutions, and exhibit vertical side walls. Proton beam micromachining (PBM) is a new technique of high potential for the manufacture of precise 3D microstructures. Recent developments have shown that metallic microstructures (nickel and copper) can be formed from these microshapes. Prototype nickel PBM resolution standards have been manufactured at the Research Centre for Nuclear Microscopy, NUS and these new standards are far superior to the 2000 mesh gold grids currently in use by many groups in terms of surface smoothness, vertical walls and edge definition. Results of beam resolution tests using the new PBM standards with the OM2000 microprobe end station/HVEE Singletron system have yielded spot sizes of 290 nm×450 nm for a 50 pA beam of 2 MeV protons.

  9. Improving the Sensitivity and Functionality of Mobile Webcam-Based Fluorescence Detectors for Point-of-Care Diagnostics in Global Health

    PubMed Central

    Rasooly, Reuven; Bruck, Hugh Alan; Balsam, Joshua; Prickril, Ben; Ossandon, Miguel; Rasooly, Avraham

    2016-01-01

    Resource-poor countries and regions require effective, low-cost diagnostic devices for accurate identification and diagnosis of health conditions. Optical detection technologies used for many types of biological and clinical analysis can play a significant role in addressing this need, but must be sufficiently affordable and portable for use in global health settings. Most current clinical optical imaging technologies are accurate and sensitive, but also expensive and difficult to adapt for use in these settings. These challenges can be mitigated by taking advantage of affordable consumer electronics mobile devices such as webcams, mobile phones, charge-coupled device (CCD) cameras, lasers, and LEDs. Low-cost, portable multi-wavelength fluorescence plate readers have been developed for many applications including detection of microbial toxins such as C. Botulinum A neurotoxin, Shiga toxin, and S. aureus enterotoxin B (SEB), and flow cytometry has been used to detect very low cell concentrations. However, the relatively low sensitivities of these devices limit their clinical utility. We have developed several approaches to improve their sensitivity presented here for webcam based fluorescence detectors, including (1) image stacking to improve signal-to-noise ratios; (2) lasers to enable fluorescence excitation for flow cytometry; and (3) streak imaging to capture the trajectory of a single cell, enabling imaging sensors with high noise levels to detect rare cell events. These approaches can also help to overcome some of the limitations of other low-cost optical detection technologies such as CCD or phone-based detectors (like high noise levels or low sensitivities), and provide for their use in low-cost medical diagnostics in resource-poor settings. PMID:27196933

  10. A new half-condensed Schiff base compound: highly selective and sensitive pH-responsive fluorescent sensor.

    PubMed

    Saha, Uday Chand; Dhara, Koushik; Chattopadhyay, Basab; Mandal, Sushil Kumar; Mondal, Swastik; Sen, Supriti; Mukherjee, Monika; van Smaalen, Sander; Chattopadhyay, Pabitra

    2011-09-02

    A new probe, 3-[(3-benzyloxypyridin-2-ylimino)methyl]-2-hydroxy-5-methylbenzaldehyde (1-H) behaves as a highly selective fluorescent pH sensor in a Britton-Robinson buffer at 25 °C. The pH titrations show a 250-fold increase in fluorescence intensity within the pH range of 4.2 to 8.3 with a pK(a) value of 6.63 which is valuable for studying many of the biological organelles.

  11. Preparation of ultra small samples for optical and microprobe analysis

    NASA Technical Reports Server (NTRS)

    Inman, C. S.

    1973-01-01

    This paper describes a simple but satisfactory new method for the preparation of tiny, varied and specialized specimens for electron or ion-microprobe analysis developed over the past five years. Microtektites, individual chondrules, single grains, blebs from lunar samples and meteoritic minerals have been prepared by this technique. A description of the preparation of these usually difficult samples from the initial mounting through the various polishing steps to their final polish is presented in detail. The procedures used to prevent any contamination of these specimens by the polishing agents and to prevent cross contamination to the other samples used for geochronology studies are presented.

  12. Electrostatic microprobe for determining charge domains on surfaces.

    PubMed

    Fletcher, Robert A

    2015-11-01

    An electrostatic microprobe was developed to measure charge on wipes and various test surfaces. The device is constructed on an optical microscope platform utilizing a computer controlled XY stage. Test surfaces can be optically imaged to identify microscopic features that can be correlated to the measured charge domain maps. The ultimate goal is to quantify charge on wipe cloths to determine the influence of electrostatic forces on wipe sampling efficiency. We found that certain wipe materials do not extensively charge while others accumulate charge by making contact with other surfaces (through the triboelectric effect). Charge domains are found to be nonuniform.

  13. Elemental mapping of biological samples using a scanning proton microprobe

    NASA Astrophysics Data System (ADS)

    Watt, F.; Grime, G. W.

    1988-03-01

    Elemental mapping using a scanning proton microprobe (SPM) can be a powerful technique for probing trace elements in biology, allowing complex interfaces to be studied in detail, identifying contamination and artefacts present in the specimen, and in certain circumstances obtaining indirect chemical information. Examples used to illustrate the advantages of the technique include the elemental mapping of growing pollen tubes, honey bee brain section, a mouse macrophage cell, human liver section exhibiting primary biliary cirrhosis, and the attack by a mildew fungus on a pea leaf.

  14. A High Resolution Microprobe Study of EETA79001 Lithology C

    NASA Technical Reports Server (NTRS)

    Schrader, Christian M.; Cohen, B. A.; Donovan, J. J.; Vicenzi, E. P.

    2010-01-01

    Antarctic meteorite EETA79001 has received substantial attention for possibly containing a component of Martian soil in its impact glass (Lithology C) [1]. The composition of Martian soil can illuminate near-surface processes such as impact gardening [2] and hydrothermal and volcanic activity [3,4]. Impact melts in meteorites represent our most direct samples of Martian regolith. We present the initial findings from a high-resolution electron microprobe study of Lithology C from Martian meteorite EETA79001. As this study develops we aim to extract details of a potential soil composition and to examine Martian surface processes using elemental ratios and correlations.

  15. Super-achromatic microprobe for ultrahigh-resolution endoscopic OCT imaging at 800 nm (Conference Presentation)

    NASA Astrophysics Data System (ADS)

    Yuan, Wu; Alemohammad, Milad; Yu, Xiaoyun; Yu, Shaoyong; Li, Xingde

    2016-03-01

    In this paper, we report a super-achromatic microprobe made with fiber-optic ball lens to enable ultrahigh-resolution endoscopic OCT imaging. An axial resolution of ~2.4 µm (in air) can be achieved with a 7-fs Ti:Sapphire laser. The microprobe has minimal astigmatism which affords a high transverse resolution of ~5.6 µm. The miniaturized microprobe has an outer diameter of ~520 µm including the encasing metal guard and can be used to image small luminal organs. The performance of the ultrahigh-resolution OCT microprobe was demonstrated by imaging rat esophagus, guinea pig esophagus, and mouse rectum in vivo.

  16. A comparison of planar, laser-induced fluorescence, and high-sensitivity interferometry techniques for gas-puff nozzle density measurements

    SciTech Connect

    Jackson, S. L.; Weber, B. V.; Mosher, D.; Phipps, D. G.; Stephanakis, S. J.; Commisso, R. J.; Qi, N.; Failor, B. H.; Coleman, P. L.

    2008-10-15

    The distribution of argon gas injected by a 12-cm-diameter triple-shell nozzle was characterized using both planar, laser-induced fluorescence (PLIF) and high-sensitivity interferometry. PLIF is used to measure the density distribution at a given time by detecting fluorescence from an acetone tracer added to the gas. Interferometry involves making time-dependent, line-integrated gas density measurements at a series of chordal locations that are then Abel inverted to obtain the gas density distribution. Measurements were made on nominally identical nozzles later used for gas-puff Z-pinch experiments on the Saturn pulsed-power generator. Significant differences in the mass distributions obtained by the two techniques are presented and discussed, along with the strengths and weaknesses of each method.

  17. A triazole Schiff base-based selective and sensitive fluorescent probe for Zn²⁺: A combined experimental and theoretical study.

    PubMed

    Yuan, Caixia; Liu, Xinyu; Wu, Yanbo; Lu, Liping; Zhu, Miaoli

    2016-02-05

    A triazole-Schiff base, 4-(5-Chloro-2-hydroxybenzylideneamino)-1H-1,2,4-triazole-5(4H)-thione (HL), exhibits the high selectivity and sensitivity for Zn(2+) in the fluorescence spectrometry over other common metal ions, especially Cd(2+) in DMSO:H2O (1:9, v/v) solution. A 1:1 binding ratio of Zn(2+)/L for the complex has been obtained by Uv-Vis titration experiments and Job's plot with the detection limit of 51 nmol/L. The coordination mode of the complex in solution was further confirmed by density functional theory (DFT) calculations. Time-dependent density functional theory (TD-DFT) calculations indicate that a chelation-enhanced fluorescence (CHEF) effect occurs in the process of detecting Zn ion.

  18. Rapid, sensitive, and selective fluorescent DNA detection using iron-based metal-organic framework nanorods: Synergies of the metal center and organic linker.

    PubMed

    Tian, Jingqi; Liu, Qian; Shi, Jinle; Hu, Jianming; Asiri, Abdullah M; Sun, Xuping; He, Yuquan

    2015-09-15

    Considerable recent attention has been paid to homogeneous fluorescent DNA detection with the use of nanostructures as a universal "quencher", but it still remains a great challenge to develop such nanosensor with the benefits of low cost, high speed, sensitivity, and selectivity. In this work, we report the use of iron-based metal-organic framework nanorods as a high-efficient sensing platform for fluorescent DNA detection. It only takes about 4 min to complete the whole "mix-and-detect" process with a low detection limit of 10 pM and a strong discrimination of single point mutation. Control experiments reveal the remarkable sensing behavior is a consequence of the synergies of the metal center and organic linker. This work elucidates how composition control of nanostructures can significantly impact their sensing properties, enabling new opportunities for the rational design of functional materials for analytical applications.

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

    PubMed

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

    2011-06-01

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

  20. A CCD-based fluorescence imaging system for real-time loop-mediated isothermal amplification-based rapid and sensitive detection of waterborne pathogens on microchips.

    PubMed

    Ahmad, Farhan; Seyrig, Gregoire; Tourlousse, Dieter M; Stedtfeld, Robert D; Tiedje, James M; Hashsham, Syed A

    2011-10-01

    Rapid, sensitive, and low-cost pathogen diagnostic systems are needed for early disease diagnosis and treatment, especially in resource-limited settings. This study reports a low-cost charge-coupled device (CCD)-based fluorescence imaging system for rapid detection of waterborne pathogens by isothermal gene amplification in disposable microchips. Fluorescence imaging capability of this monochromatic CCD camera is evaluated by optimizing the gain, offset, and exposure time. This imaging system is validated for 12 virulence genes of major waterborne pathogens on cyclic olefin polymer (COP) microchips, using SYTO-82 dye and real time fluorescence loop-mediated isothermal amplification referred here as microRT(f)-LAMP. Signal-to-noise ratio (SNR) and threshold time (Tt) of microRT(f)-LAMP assays are compared with those from a commercial real-time polymerase chain reaction (PCR) instrument. Applying a CCD exposure of 5 s to 10(5) starting DNA copies of microRT(f)-LAMP assays increases the SNR by 8-fold and reduces the Tt by 9.8 min in comparison to a commercial real-time PCR instrument. Additionally, single copy level sensitivity for Campylobacter jejuni 0414 gene is obtained for microRT(f)-LAMP with a Tt of 19 min, which is half the time of the commercial real-time PCR instrument. Due to the control over the exposure time and the wide field imaging capability of CCD, this low-cost fluorescence imaging system has the potential for rapid and parallel detection of pathogenic microorganisms in high throughput microfluidic chips.

  1. Volume labeling with Alexa Fluor dyes and surface functionalization of highly sensitive fluorescent silica (SiO2) nanoparticles

    NASA Astrophysics Data System (ADS)

    Wang, Wei; Nallathamby, Prakash D.; Foster, Carmen M.; Morrell-Falvey, Jennifer L.; Mortensen, Ninell P.; Doktycz, Mitchel J.; Gu, Baohua; Retterer, Scott T.

    2013-10-01

    A new synthesis approach is described that allows the direct incorporation of fluorescent labels into the volume or body of SiO2 nanoparticles. In this process, fluorescent Alexa Fluor dyes with different emission wavelengths were covalently incorporated into the SiO2 nanoparticles during their formation by the hydrolysis of tetraethoxysilane. The dye molecules were homogeneously distributed throughout the SiO2 nanoparticles. The quantum yields of the Alexa Fluor volume-labeled SiO2 nanoparticles were much higher than nanoparticles labeled using conventional organic dyes. The size of the resulting nanoparticles was controlled using microemulsion reaction media with sizes in the range of 20-100 nm and a polydispersity of <15%. In comparison with conventional surface tagged particles created by post-synthesis modification, this process maintains the physical and surface chemical properties that have the most pronounced effect on colloidal stability and interactions with their surroundings. These volume-labeled nanoparticles have proven to be extremely robust, showing excellent signal strength, negligible photobleaching, and minimal loss of functional organic components. The native or ``free'' surface of the volume-labeled particles can be altered to achieve a specific surface functionality without altering fluorescence. Their utility was demonstrated for visualizing the association of surface-modified fluorescent particles with cultured macrophages. Differences in particle agglomeration and cell association were clearly associated with differences in observed nanoparticle toxicity. The capacity to maintain particle fluorescence while making significant changes to surface chemistry makes these particles extremely versatile and useful for studies of particle agglomeration, uptake, and transport in environmental and biological systems.A new synthesis approach is described that allows the direct incorporation of fluorescent labels into the volume or body of SiO2

  2. Volume Labeling with Alexa-Fluor Dyes and Surface Functionalization of Highly Sensitive Fluorescent SiO2 Nanoparticles

    SciTech Connect

    Wang, Wei; Foster, Carmen M; Morrell-Falvey, Jennifer L; Nallathamby, Prakash D; Mortensen, Ninell P; Doktycz, Mitchel John; Gu, Baohua; Retterer, Scott T; Gu, Baohua

    2013-01-01

    A new synthesis approach is described that allows the direct incorporation of fluorescent labels into the volume or body of SiO2 nanoparticles. In this process, fluorescent Alexa Fluor dyes with different emission wavelengths were covalently incorporated into the SiO2 nanoparticles during their formation by the hydrolysis of tetraethoxysilane. The dye molecules were homogeneously distributed throughout the SiO2 nanoparticles. The quantum yields of the Alexa Fluor volume-labeled SiO2 nanoparticles were much higher than nanoparticles labeled using conventional organic dyes. The size of the resulting nanoparticles was controlled using microemulsion reaction media with sizes in the range of 20-100 nm and a polydispersity of <15%. In comparison with conventional surface tagged particles created by post-synthesis modification, this process maintains the physical and surface chemical properties that have the most pronounced effect on colloidal stability and interactions with their surroundings. These volume-labeled nanoparticles have proven to be extremely robust, showing excellent signal strength, negligible photobleaching, and minimal loss of functional organic components. The native or free surface of the volume-labeled particles can be altered to achieve a specific surface functionality without altering fluorescence. Their utility was demonstrated for visualizing the association of surface modified fluorescent particles with cultured macrophages. Differences in particle agglomeration and cell association were clearly associated with differences in observed nanoparticle toxicity. The capacity to maintain particle fluorescence while making significant changes to surface chemistry makes these particles extremely versatile and useful for studies of particle agglomeration, uptake, and transport in environmental and biological systems.

  3. Volume labeling with Alexa Fluor dyes and surface functionalization of highly sensitive fluorescent silica (SiO2) nanoparticles.

    PubMed

    Wang, Wei; Nallathamby, Prakash D; Foster, Carmen M; Morrell-Falvey, Jennifer L; Mortensen, Ninell P; Doktycz, Mitchel J; Gu, Baohua; Retterer, Scott T

    2013-11-07

    A new synthesis approach is described that allows the direct incorporation of fluorescent labels into the volume or body of SiO2 nanoparticles. In this process, fluorescent Alexa Fluor dyes with different emission wavelengths were covalently incorporated into the SiO2 nanoparticles during their formation by the hydrolysis of tetraethoxysilane. The dye molecules were homogeneously distributed throughout the SiO2 nanoparticles. The quantum yields of the Alexa Fluor volume-labeled SiO2 nanoparticles were much higher than nanoparticles labeled using conventional organic dyes. The size of the resulting nanoparticles was controlled using microemulsion reaction media with sizes in the range of 20-100 nm and a polydispersity of <15%. In comparison with conventional surface tagged particles created by post-synthesis modification, this process maintains the physical and surface chemical properties that have the most pronounced effect on colloidal stability and interactions with their surroundings. These volume-labeled nanoparticles have proven to be extremely robust, showing excellent signal strength, negligible photobleaching, and minimal loss of functional organic components. The native or "free" surface of the volume-labeled particles can be altered to achieve a specific surface functionality without altering fluorescence. Their utility was demonstrated for visualizing the association of surface-modified fluorescent particles with cultured macrophages. Differences in particle agglomeration and cell association were clearly associated with differences in observed nanoparticle toxicity. The capacity to maintain particle fluorescence while making significant changes to surface chemistry makes these particles extremely versatile and useful for studies of particle agglomeration, uptake, and transport in environmental and biological systems.

  4. A novel lab-on-chip platform with integrated solid phase PCR and Supercritical Angle Fluorescence (SAF) microlens array for highly sensitive and multiplexed pathogen detection.

    PubMed

    Hung, Tran Quang; Chin, Wai Hoe; Sun, Yi; Wolff, Anders; Bang, Dang Duong

    2017-04-15

    Solid-phase PCR (SP-PCR) has become increasingly popular for molecular diagnosis and there have been a few attempts to incorporate SP-PCR into lab-on-a-chip (LOC) devices. However, their applicability for on-line diagnosis is hindered by the lack of sensitive and portable on-chip optical detection technology. In this paper, we addressed this challenge by combining the SP-PCR with super critical angle fluorescence (SAF) microlens array embedded in a microchip. We fabricated miniaturized SAF microlens array as part of a microfluidic chamber in thermoplastic material and performed multiplexed SP-PCR directly on top of the SAF microlens array. Attribute to the high fluorescence collection efficiency of the SAF microlens array, the SP-PCR assay on the LOC platform demonstrated a high sensitivity of 1.6 copies/µL, comparable to off-chip detection using conventional laser scanner. The combination of SP-PCR and SAF microlens array allows for on-chip highly sensitive and multiplexed pathogen detection with low-cost and compact optical components. The LOC platform would be widely used as a high-throughput biosensor to analyze food, clinical and environmental samples.

  5. A LSO β microprobe for measuring input functions for quantitative small animal PET

    NASA Astrophysics Data System (ADS)

    Maramraju, S.; Stoll, S.; Woody, C.; Schlyer, D.; Schiffer, W.; Lee, D.; Dewey, S.; Vaska, P.

    2007-02-01

    A miniature scintillation microprobe has been developed to measure the input function in live rodents for use in longitudinal, quantitative PET studies. The probe consists of a small lutetium oxyorthosilicate (LSO) crystal measuring typically 0.3-0.5 mm diameter ×0.5-2 mm in length that is used to directly detect positrons in the blood or tissue. The probe has a sensitivity of 10-30 Hz/μCi/cm 3 and is primarily sensitive to short-range positrons emitted by labeled radiotracers in the blood. The sensitivity to γ-ray background can be minimized using a variable threshold in the readout to discriminate between positrons and γ's. The probe was implanted in one of the tail veins of a Sprague-Dawley rat and the input function was measured for the injection of 0.8 mCi of FDG in the other tail vein. The probe exhibits a fast time response that is able to quickly and accurately measure the concentration of 18F circulating in the bloodstream. Additional tests were also carried out to study the probe's sensitivity to γ-ray background.

  6. pH-Sensitive self-assembling nanoparticles for tumor near-infrared fluorescence imaging and chemo-photodynamic combination therapy

    NASA Astrophysics Data System (ADS)

    Hou, Wenxiu; Zhao, Xin; Qian, Xiaoqing; Pan, Fei; Zhang, Chunlei; Yang, Yuming; de La Fuente, Jesús Martínez; Cui, Daxiang

    2015-12-01

    The development of visual tumor theranostic nanoparticles has become a great challenge. In this study, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) was conjugated to acid-sensitive cis-aconitic anhydride-modified doxorubicin (CAD) to obtain pH-sensitive anti-tumor prodrug nanoparticles (TCAD NPs) via self-assembling. Subsequently, the photosensitizer chlorin e6 (Ce6) was loaded into the resulting prodrug nanoparticles to prepare a novel tumor near-infrared fluorescence imaging and chemo-photodynamic combination therapy system (TCAD@Ce6 NPs). An accelerated release of doxorubicin (DOX) and chlorin e6 (Ce6) from the TCAD@Ce6 NPs could be achieved due to the hydrolysis of the acid-sensitive amide linker under mild acidic conditions (pH = 5.5). An in vitro experiment showed that A549 lung cancer cells exhibited a significantly higher uptake of DOX and Ce6 by using our delivery system than the free form of DOX and Ce6. An in vivo experiment showed that TCAD@Ce6 NPs displayed better tumor targeting gathering through the enhanced permeability and retention (EPR) effect than free Ce6, thus improving fluorescence imaging. Moreover, the chemo-photodynamic combination therapy of TCAD@Ce6 NPs combined with near-infrared laser irradiation was confirmed to be capable of inducing high apoptosis and necrosis of tumor cells (A549) in vitro and to display a significantly higher tumor growth suppression in the A549 lung cancer-bearing mice model. Furthermore, compared with exclusive chemotreatment (DOX) or photodynamic treatment (Ce6), our system showed enhanced therapeutic effects both in vitro and in vivo. In conclusion, the high performance TCAD@Ce6 NPs can be used as a promising NIR fluorescence imaging and highly effective chemo-photodynamic system for theranostics of lung cancer, etc. in the near future.The development of visual tumor theranostic nanoparticles has become a great challenge. In this study, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) was

  7. Nuclear microprobe applications to radioactive waste management basic research

    NASA Astrophysics Data System (ADS)

    Trocellier, P.; Badillo, V.; Barré, N.; Bois, L.; Cachoir, C.; Gallien, J. P.; Guilbert, S.; Mercier, F.; Tiffreau, C.

    1999-10-01

    Radioactive waste management is one of the major technical and scientific challenge to be solved by industrialized countries near the beginning of the 21st century. Relevant questions arise about the extrapolation of the long term-behavior of materials from waste package, engineered barriers and near field repository. Whatever the strategical option might be, wet atmosphere or water intrusion through the different barriers constitute the two main remobilization factors for radionuclides in the geosphere and the biosphere. The study of solid alteration processes and elemental sorption phenomena on mineral surfaces is one of the most efficient basic research approaches to assess the long term performance of waste materials. Ion beam analysis and more recently nuclear microprobe techniques have been applied to investigate exchange mechanisms near representative solid/liquid interfaces such as glass/deionized water, uranium dioxide/granitic or clay water or mineral surface/aqueous solution doped with chemical elements analogue to actinide or fission products. This paper intends to describe the different works that have been carried out in Saclay using the nuclear microprobe facility. The coupling of μRBS, μPIXE and μNRA permits to determine the evolution of the surface composition induced by chemical reactions involved. Complementary observation of solid morphology and solution analysis allows to obtain a complete elemental balance on exchange processes.

  8. Applications of the nuclear microprobe in planetary science

    NASA Astrophysics Data System (ADS)

    Vis, R. D.

    1997-07-01

    Nuclear microprobes have been used in a variety of studies on extra-terrestrial materials. Although by far the most used analytical technique is micro-PIXE, valuable contributions have also been given to planetary science using other methods available among the suite of analytical techniques provided by the microprobe. Also a few studies of the application of synchrotron radiation to planetary science has been published. Research aims are either to get a full analysis of very small objects such as cosmic dust or to extract elemental profiles over areas of interest. In the latter case, these distributions may give insight into the temperature history of the objects studied. In this way single crystals, chondrules in ordinary chondrites but also phase transitions in iron-meteorites have been investigated. Being by far the oldest objects available for research and being conserved for billions of years without serious wearing and erosion as would happen on earth, their detailed studies provide knowledge about the early history of the solar system and on primary geological processes.

  9. A Comparison of the Capability of Sensitivity Level 3 and Sensitivity Level 4 Fluorescent Penetrants to Detect Fatigue Cracks in Various Metals

    NASA Technical Reports Server (NTRS)

    Parker, Bradford H.

    2011-01-01

    In April 2008, NASA-STD-5009 established a requirement that only sensitivity level 4 penetrants are acceptable for NASA Standard Level liquid penetrant inspections. Having NASA contractors change existing processes or perform demonstration tests to certify sensitivity level 3 penetrants posed a potentially huge cost to the Agency. This study was conducted to directly compare the probability of detection (POD) of sensitivity level 3 and level 4 penetrants using both Method A and Method D inspection processes. POD demonstration tests were performed on 6061-Al, Haynes 188 and Ti-6Al-4V crack panel sets. The study results strongly support the conclusion that sensitivity level 3 penetrants are acceptable for NASA Standard Level inspections.

  10. pH-Sensitive self-assembling nanoparticles for tumor near-infrared fluorescence imaging and chemo-photodynamic combination therapy.

    PubMed

    Hou, Wenxiu; Zhao, Xin; Qian, Xiaoqing; Pan, Fei; Zhang, Chunlei; Yang, Yuming; de la Fuente, Jesús Martínez; Cui, Daxiang

    2016-01-07

    The development of visual tumor theranostic nanoparticles has become a great challenge. In this study, d-α-tocopheryl polyethylene glycol 1000 succinate (TPGS) was conjugated to acid-sensitive cis-aconitic anhydride-modified doxorubicin (CAD) to obtain pH-sensitive anti-tumor prodrug nanoparticles (TCAD NPs) via self-assembling. Subsequently, the photosensitizer chlorin e6 (Ce6) was loaded into the resulting prodrug nanoparticles to prepare a novel tumor near-infrared fluorescence imaging and chemo-photodynamic combination therapy system (TCAD@Ce6 NPs). An accelerated release of doxorubicin (DOX) and chlorin e6 (Ce6) from the TCAD@Ce6 NPs could be achieved due to the hydrolysis of the acid-sensitive amide linker under mild acidic conditions (pH = 5.5). An in vitro experiment showed that A549 lung cancer cells exhibited a significantly higher uptake of DOX and Ce6 by using our delivery system than the free form of DOX and Ce6. An in vivo experiment showed that TCAD@Ce6 NPs displayed better tumor targeting gathering through the enhanced permeability and retention (EPR) effect than free Ce6, thus improving fluorescence imaging. Moreover, the chemo-photodynamic combination therapy of TCAD@Ce6 NPs combined with near-infrared laser irradiation was confirmed to be capable of inducing high apoptosis and necrosis of tumor cells (A549) in vitro and to display a significantly higher tumor growth suppression in the A549 lung cancer-bearing mice model. Furthermore, compared with exclusive chemotreatment (DOX) or photodynamic treatment (Ce6), our system showed enhanced therapeutic effects both in vitro and in vivo. In conclusion, the high performance TCAD@Ce6 NPs can be used as a promising NIR fluorescence imaging and highly effective chemo-photodynamic system for theranostics of lung cancer, etc. in the near future.

  11. Highly sensitive dual-mode fluorescence detection of lead ion in water using aggregation-induced emissive polymers.

    PubMed

    Saha, Sukanta Kumar; Ghosh, Khama Rani; Gao, Jian Ping; Wang, Zhi Yuan

    2014-09-01

    A series of fluorene-based conjugated polymers containing the aggregation-induced emissive (AIE)-active tetraphenylethene and dicarboxylate pseudocrown as a receptor exhibits a unique dual-mode sensing ability for selective detection of lead ion in water. Fluorescence turn-off and turn-on detections are realized in 80%-90% and 20% water in tetrahydrofuran (THF), respectively, for lead ion with a concentration as low as 10(-8) m.

  12. A highly sensitive and selective fluorescent sensor for detection of Al(3+) using a europium(III) quinolinecarboxylate.

    PubMed

    Xu, Wentao; Zhou, Youfu; Huang, Decai; Su, Mingyi; Wang, Kun; Hong, Maochun

    2014-07-07

    Eu2PQC6 has been developed to detect Al(3+) by monitoring the quenching of the europium-based emission, with the lowest detection limit of ∼32 pM and the quantitative detection range to 150 μM. Eu2PQC6 is the first ever example that the europium(III) complex serves as an Al(3+) fluorescent sensor based on "competition-displacement" mode.

  13. TU-G-207-03: High Spatial Resolution and High Sensitivity X-Ray Fluorescence Imaging

    SciTech Connect

    Xing, L.

    2015-06-15

    Last few years has witnessed the development of novel of X-ray imaging modalities, such as spectral CT, phase contrast CT, and X-ray acoustic/fluorescence/luminescence imaging. This symposium will present the recent advances of these emerging X-ray imaging modalities and update the attendees with knowledge in various related topics, including X-ray photon-counting detectors, X-ray physics underlying the emerging applications beyond the traditional X-ray imaging, image reconstruction for the novel modalities, characterization and evaluation of the systems, and their practical implications. In addition, the concept and practical aspects of X-ray activatable targeted nanoparticles for molecular X-ray imaging will be discussed in the context of X-ray fluorescence and luminescence CT. Learning Objectives: Present background knowledge of various emerging X-ray imaging techniques, such as spectral CT, phase contrast CT and X-ray fluorescence/luminescence CT. Discuss the practical need, technical aspects and current status of the emerging X-ray imaging modalities. Describe utility and future impact of the new generation of X-ray imaging applications.

  14. DNA derived fluorescent bio-dots for sensitive detection of mercury and silver ions in aqueous solution

    NASA Astrophysics Data System (ADS)

    Song, Ting; Zhu, Xuefeng; Zhou, Shenghai; Yang, Guang; Gan, Wei; Yuan, Qunhui

    2015-08-01

    Inspired by the high affinity between heavy metal ions and bio-molecules as well as the low toxicity of carbon-based quantum dots, we demonstrated the first application of a DNA derived carbonaceous quantum dots, namely bio-dots, in metal ion sensing. The present DNA-derived bio-dots contain graphitic carbon layers with 0.242 nm lattice fringes, exhibit excellent fluorescence property and can be obtained via a facile hydrothermal preparation procedure. Hg(II) and Ag(I) are prone to be captured by the bio-dots due to the existence of residual thymine (T) and cytosine (C) groups, resulting in a quenched fluorescence while other heavy metal ions would cause negligible changes on the fluorescent signals of the bio-dots. The bio-dots could be used as highly selective toxic-free biosensors, with two detecting linear ranges of 0-0.5 μM and 0.5-6 μM for Hg(II) and one linear range of 0-10 μM for Ag(I). The detection limits (at a signal-to-noise ratio of 3) were estimated to be 48 nM for Hg(II) and 0.31 μM for Ag(I), respectively. The detection of Hg(II) and Ag(I) could also be realized in the real water sample analyses, with satisfying recoveries ranging from 87% to 100%.

  15. Environment-sensitive quinolone demonstrating long-lived fluorescence and unusually slow excited-state intramolecular proton transfer kinetics

    NASA Astrophysics Data System (ADS)

    Zamotaiev, O. M.; Shvadchak, V.; Sych, T. P.; Melnychuk, N. A.; Yushchenko, D.; Mely, Y.; Pivovarenko, V. G.

    2016-09-01

    A new small fluorescent dye based on 3-hydroxybenzo[g]quinolone, a benzo-analogue of Pseudomonas quinolone signal species, has been synthesized. The dye demonstrates interesting optical properties, with absorption in the visible region, two band emission due to an excited-state intramolecular proton transfer (ESIPT) reaction and high fluorescence quantum yield in both protic and aprotic media. Time-resolved fluorescence spectroscopy shows that the ESIPT reaction time is unusually long (up to 8 ns), indicating that both forward and backward ESIPT reactions are very slow in comparison to other 3-hydroxyquinolones. In spite of these slow rate constants, the ESIPT reaction was found to show a reversible character as a result of the very long lifetimes of both N* and T* forms (up to 16 ns). The ESIPT reaction rate is mainly controlled by the hydrogen bond donor ability in protic solvents and the polarity in aprotic solvents. Using large unilamellar vesicles and giant unilamellar vesicles of different lipid compositions, the probe was shown to preferentially label liquid disordered phases.

  16. Facile synthesis of boron- and nitride-doped MoS2 nanosheets as fluorescent probes for the ultrafast, sensitive, and label-free detection of Hg(2+).

    PubMed

    Liu, Xiaojia; Li, Liping; Wei, Yuanjie; Zheng, Yizhi; Xiao, Qian; Feng, Bo

    2015-07-07

    Bulk MoS2, a prototypical transition metal chalcogenide material, is an indirect band gap semiconductor with negligible photoluminescence. In this study, we have developed, for the first time, a simple and low-cost synthetic strategy to prepare boron- and nitrogen-doped MoS2 (B,N-MoS2) nanosheets. Through boron and nitrogen doping, the band gap of MoS2 increases from 1.20 eV to 1.61 eV, and the obtained B,N-MoS2 nanosheets exhibit an enhanced fluorescence. The B,N-MoS2 nanosheets can be used as a green and facile sensing platform for label-free detection of Hg(2+) because of their high sensitivity and selectivity toward Hg(2+). In addition, detection can be easily accomplished through one-step rapid (within 2 min) operation, with a limit as low as 1 nM. This study demonstrates that the introduction of boron and nitrogen elements into ultrathin MoS2 nanosheets for enhanced fluorescence properties is feasible through a facile and general preparation strategy and may also offer a unique idea as a potential way to design more efficient MoS2-based sensors and fluorescent materials.

  17. Simple and sensitive detection method for Cobalt(II) in water using CePO4:Tb3+ nanocrystals as fluorescent probes.

    PubMed

    Chen, Hongqi; Yuan, Fei; Xu, Juan; Zhang, Yiyang; Wu, Yong; Wang, Lun

    2013-04-15

    A simple and sensitive method for detecting cobalt by synchronous fluorescence spectrometry technique with a novel fluorescence probe CePO4:Tb(3+) has been developed. CePO4:Tb(3+) nanocrystals were synthesized in aqueous solutions and characterized by transmission electron microscopy, electron diffraction pattern spectroscopy and spectrofluorometry. When Δλ=210 nm, the selected synchronous fluorescence is produced at 284 nm. CePO4:Tb(3+) nanocrystals were negatively charged under weakly basic conditions (pH=8.2), which can interact with Co(2+) via electrostatic interaction. Moreover, there is the spectrum overlap between the emission wavelength of CePO4:Tb(3+) NCs and the absorbance of Co(2+). So the energy transfer would occur, leading to the quenching phenomenon. The quenching equation of the system was agreed with the Stern-Volmer equation. The linear range and detection limit of Co(2+) were 5-1.8 μM and 3.5 nM, respectively. The method is successfully applied to the quantification of Co(2+) in water samples.

  18. Efficient ensemble system based on the copper binding motif for highly sensitive and selective detection of cyanide ions in 100% aqueous solutions by fluorescent and colorimetric changes.

    PubMed

    Jung, Kwan Ho; Lee, Keun-Hyeung

    2015-09-15

    A peptide-based ensemble for the detection of cyanide ions in 100% aqueous solutions was designed on the basis of the copper binding motif. 7-Nitro-2,1,3-benzoxadiazole-labeled tripeptide (NBD-SSH, NBD-SerSerHis) formed the ensemble with Cu(2+), leading to a change in the color of the solution from yellow to orange and a complete decrease of fluorescence emission. The ensemble (NBD-SSH-Cu(2+)) sensitively and selectively detected a low concentration of cyanide ions in 100% aqueous solutions by a colorimetric change as well as a fluorescent change. The addition of cyanide ions instantly removed Cu(2+) from the ensemble (NBD-SSH-Cu(2+)) in 100% aqueous solutions, resulting in a color change of the solution from orange to yellow and a "turn-on" fluorescent response. The detection limits for cyanide ions were lower than the maximum allowable level of cyanide ions in drinking water set by the World Health Organization. The peptide-based ensemble system is expected to be a potential and practical way for the detection of submicromolar concentrations of cyanide ions in 100% aqueous solutions.

  19. Ultrasound-assisted dispersive liquid-liquid microextraction combined with high-performance liquid chromatography-fluorescence detection for sensitive determination of biogenic amines in rice wine samples.

    PubMed

    Huang, Ke-Jing; Wei, Cai-Yun; Liu, Wei-Li; Xie, Wan-Zhen; Zhang, Jun-Feng; Wang, Wei

    2009-09-18

    Ultrasound-assisted dispersive liquid-liquid microextraction coupled with high-performance liquid chromatography-fluorescence detection was used for the extraction and determination of three biogenic amines including octopamine, tyramine and phenethylamine in rice wine samples. Fluorescence probe 2,6-dimethyl-4-quinolinecarboxylic acid N-hydroxysuccinimide ester was applied for derivatization of biogenic amines. Acetonitrile and 1-octanol were used as disperser solvent and extraction solvent, respectively. Extraction conditions including the type of extraction solvent, the volume of extraction solvent, ultrasonication time and centrifuging time were optimized. After extraction and centrifuging, analyte was injected rapidly into high-performance liquid chromatography and then detected with fluorescence. The calibration graph of the proposed method was linear in the range of 5-500 microg mL(-1) (octopamine and tyramine) and 0.025-2.5 microg mL(-1) (phenethylamine). The relative standard deviations were 2.4-3.2% (n=6) and the limits of detection were in the range of 0.02-5 ng mL(-1). The method was applied to analyze the rice wine samples and spiked recoveries in the range of 95.42-104.56% were obtained. The results showed that ultrasound-assisted dispersive liquid-liquid microextraction was a very simple, rapid, sensitive and efficient analytical method for the determination of trace amount of biogenic amines.

  20. RABBIT: an electron microprobe data-reduction program using empirical corrections

    USGS Publications Warehouse

    Goff, Fraser E.

    1977-01-01

    RABBIT is a FORTRAN IV computer Program that uses Bence-Albee empirical corrections for the reduction of electron microprobe data of silicates, oxides, sulphates, carbonates, and phosphates. RABBIT efficiently reduces large volumes of data collected on 3-11 channel microprobes.

  1. Molecular microanalysis of pathological specimens in situ with a laser-Raman microprobe.

    PubMed

    Abraham, J L; Etz, E S

    1979-11-09

    A laser-Raman microprobe has been used to identify microscopic inclusions of silicone polymer in standard paraffin sections of lymph node. This example of organic chemical microanalysis in situ in pathological tissue represents an extension of microanalytical capabilities from elemental analysis, performed with electron and ion microprobes, to compound-specific molecular microanalysis.

  2. Motor-based microprobe powered by bio-assembled catalase for motion detection of DNA.

    PubMed

    Xie, Yuzhe; Fu, Shizhe; Wu, Jie; Lei, Jianping; Ju, Huangxian

    2017-01-15

    A motor-based microprobe is proposed using a tubular microengine powered by bio-assembled enzyme as catalyst and exploited for washing-free detection of DNA through motion readout. The microprobe is fabricated by assembling a catalase layer on the inner surface of poly(3,4-ethylenedioxythiophene)/Au (PEDOT/Au) microtube through DNA conjugate, which is responsible for the biocatalytic bubble propulsion. The sensing concept of the microprobe relies on the target-induced release of catalase through the DNA strand-replacement hybridization, which decreases the amount of enzyme assembled on microtube to slow down the movement of the microprobe. Therefore, the motion speed is negatively correlated with the target concentration. At the optimal conditions, the microprobe can conveniently distinguish the concentration of specific DNA in a range of 0.5-10µM without any washing and separation step. This microprobe can be prepared in batch with good reproducibility and stability, and its motion speed can be conveniently visualized by optical microscope. The proposed motor-based microprobe and its dynamic sensing method provide a novel platform for the development of intelligent microprobe and clinical diagnostic strategy.

  3. A novel graphene-based label-free fluorescence `turn-on' nanosensor for selective and sensitive detection of phosphorylated species in biological samples and living cells

    NASA Astrophysics Data System (ADS)

    Ke, Yaotang; Garg, Bhaskar; Ling, Yong-Chien

    2016-02-01

    A novel label-free fluorescence `turn-on' nanosensor has been developed for highly selective and sensitive detection of phosphorylated species (Ps) in biological samples and living cells. The design strategy relies on the use of Ti4+-immobilized polydopamine (PDA) coated reduced graphene oxide (rGO@PDA-Ti4+) that serves as an attractive platform to bind riboflavin 5'-monophosphate molecules (FMNs) through ion-pair interactions between phosphate groups and Ti4+. The as-prepared rGO@PDA-Ti4+-FMNs (nanosensor), fluoresce only weakly due to the ineffective Förster resonance energy transfer between the FMNs and rGO@PDA-Ti4+. The experimental findings revealed that the microwave-assisted interaction of the nanosensor with α-, β-casein, ovalbumin, human serum, non-fat milk, egg white, and living cells (all containing Ps) releases FMNs (due to the high formation constant between phosphate groups and Ti4+), leading to an excellent fluorescence `turn-on' response. The fluorescence spectroscopy, confocal microscopy, and MALDI-TOF MS spectrometry were used to detect Ps both qualitatively and quantitatively. Under the optimized conditions, the nanosensor showed a detection limit of ca. 118.5, 28.9, and 54.8 nM for the tryptic digests of α-, β-casein and ovalbumin, respectively. Furthermore, the standard addition method was used as a bench-mark proof for phosphopeptide quantification in egg white samples. We postulate that the present quantitative assay for Ps holds tremendous potential and may pave the way to disease diagnostics in the near future.A novel label-free fluorescence `turn-on' nanosensor has been developed for highly selective and sensitive detection of phosphorylated species (Ps) in biological samples and living cells. The design strategy relies on the use of Ti4+-immobilized polydopamine (PDA) coated reduced graphene oxide (rGO@PDA-Ti4+) that serves as an attractive platform to bind riboflavin 5'-monophosphate molecules (FMNs) through ion-pair interactions

  4. Rapid Preparation of Released N-Glycans for HILIC Analysis Using a Labeling Reagent that Facilitates Sensitive Fluorescence and ESI-MS Detection.

    PubMed

    Lauber, Matthew A; Yu, Ying-Qing; Brousmiche, Darryl W; Hua, Zhengmao; Koza, Stephan M; Magnelli, Paula; Guthrie, Ellen; Taron, Christopher H; Fountain, Kenneth J

    2015-01-01

    N-glycosylation of proteins is now routinely characterized and monitored because of its significance to the detection of disease states and the manufacturing of biopharmaceuticals. At the same time, hydrophilic interaction chromatography (HILIC) has emerged as a powerful technology for N-glycan profiling. Sample preparation techniques for N-glycan HILIC analyses have however tended to be laborious or require compromises in sensitivity. To address these shortcomings, we have developed an N-glycan labeling reagent that provides enhanced fluorescence response and MS sensitivity for glycan detection and have also simplified the process of preparing a sample for analysis. The developed labeling reagent rapidly reacts with glycosylamines upon their release from glycoproteins. Within a 5 min reaction, enzymatically released N-glycans are labeled with this reagent comprised of an NHS-carbamate reactive group, a quinoline fluorophore, and a tertiary amine for enhancing ESI+ MS ionization. To further expedite the released N-glycan sample preparation, rapid tagging has been integrated with a fast PNGase F deglycosylation procedure that achieves complete deglycosylation of a diverse set of glycoproteins in approximately 10 min. Moreover, a technique for HILIC-SPE of the labeled glycans has been developed to provide quantitative recovery and facilitate immediate HILIC analysis of the prepared samples. The described approach makes it possible to quickly prepare N-glycan samples and to incorporate the use of a fluorescence and MS sensitivity enhancing labeling reagent. In demonstration of these new capabilities, we have combined the developed sample preparation techniques with UHPLC HILIC chromatography and high sensitivity mass spectrometry to thoroughly detail the N-glycan profile of a monoclonal antibody.

  5. Sensitive and rapid detection of endogenous hydrogen sulfide distributing in different mouse viscera via a two-photon fluorescent probe.

    PubMed

    Chen, Qian; Yang, Jinfeng; Li, Yinhui; Zheng, Jing; Yang, Ronghua

    2015-10-08

    Development of efficient methods for detection of endogenous H2S in living cells and tissues is of considerable significance for better understanding the biological and pathological functions of H2S. Two-photon (TP) fluorescent probes are favorable as powerful molecular tools for studying physiological process due to its non-invasiveness, high spatiotemporal resolution and deep-tissues imaging. Up to date, several TP probes for intracellular H2S imaging have been designed, but real-time imaging of endogenous H2S-related biological processes in tissues is hampered due to low sensitivity, long response time and interference from other biothiols. To address this issue, we herein report a novel two-photon fluorescent probe (TPP-H2S) for highly sensitive and fast monitoring and imaging H2S levels in living cells and tissues. In the presence of H2S, it exhibits obviously improved sensitivity (LOD: 0.12 μM) and fast response time (about 2 min) compared with the reported two-photon H2S probes. With two-photon excitation, TPP-H2S displays high signal-to-noise ratio and sensitivity even no interference in cell growth media. As further application, TPP-H2S is applied for fast imaging of H2S in living cells and different fresh tissues by two-photon confocal microscope. Most importantly we first measured the endogenous H2S level in different viscera by vivisection and found that the distribution of endogenous H2S mostly in brain, liver and lung. The excellent sensing properties of TPP-H2S make it a practically useful tool for further studying biological roles of H2S.

  6. Site-specific fluorescence dynamics in an RNA ‘thermometer’ reveals the role of ribosome binding in its temperature-sensitive switch function

    PubMed Central

    Narayan, Satya; Kombrabail, Mamta H.; Das, Sudipta; Singh, Himanshu; Chary, Kandala V. R.; Rao, Basuthkar J.; Krishnamoorthy, Guruswamy

    2015-01-01

    RNA thermometers control the translation of several heat shock and virulence genes by their temperature-sensitive structural transitions. Changes in the structure and dynamics of MiniROSE RNA, which regulates translation in the temperature range of 20–45°C, were studied by site specifically replacing seven adenine residues with the fluorescent analog, 2-aminopurine (2-AP), one at a time. Dynamic fluorescence observables of 2-AP-labeled RNAs were compared in their free versus ribosome-bound states for the first time. Noticeably, position dependence of fluorescence observables, which was prominent at 20°C, was persistent even at 45ºC, suggesting the persistence of structural integrity up to 45ºC. Interestingly, position-dependent dispersion of fluorescence lifetime and quenching constant at 45°C was ablated in ribosome-bound state, when compared to those at 20°C, underscoring loss of structural integrity at 45°C, in ribosome-bound RNA. Significant increase in the value of mean lifetime for 2-AP corresponding to Shine–Dalgarno sequences, when the temperature was raised from 20 to 45°C, to values seen in the presence of urea at 45°C was a strong indicator of melting of the 3D structure of MiniROSE RNA at 45°C, only when it was ribosome bound. Taken all together, we propose a model where we invoke that ribosome binding of the RNA thermometer critically regulates temperature sensing functions in MiniROSE RNA. PMID:25477380

  7. Europium-decorated graphene quantum dots as a fluorescent probe for label-free, rapid and sensitive detection of Cu(2+) and L-cysteine.

    PubMed

    Lin, Liping; Song, Xinhong; Chen, Yiying; Rong, Mingcong; Wang, Yiru; Zhao, Li; Zhao, Tingting; Chen, Xi

    2015-09-03

    In this work, europium-decorated graphene quantum dots (Eu-GQDs) were prepared by treating three-dimensional Eu-decorated graphene (3D Eu-graphene) via a strong acid treatment. Various characterizations revealed that Eu atoms were successfully complexed with the oxygen functional groups on the surface of graphene quantum dots (GQDs) with the atomic ratio of 2.54%. Compared with Eu free GQDs, the introduction of Eu atoms enhanced the electron density and improved the surface chemical activities of Eu-GQDs. Therefore, the obtained Eu-GQDs were used as a novel "off-on" fluorescent probe for the label-free determination of Cu(2+) and l-cysteine (L-Cys) with high sensitivity and selectivity. The fluorescence intensity of Eu-GQDs was quenched in the presence of Cu(2+) owing to the coordination reaction between Cu(2+) and carboxyl groups on the surface of the Eu-GQDs. The fluorescence intensity of Eu-GQDs recovered with the subsequent addition of L-Cys because of the strong affinity of Cu(2+) to L-Cys via the Cu-S bond. The experimental results showed that the fluorescence variation of the proposed approach had a good linear relationship in the range of 0.1-10 μM for Cu(2+) and 0.5-50 μM for L-Cys with corresponding detection limits of 0.056 μM for Cu(2+) and 0.31 μM for L-Cys. The current approach also displayed a special response to Cu(2+) and L-Cys over the other co-existing metal ions and amino acids, and the results obtained from buffer-diluted serum samples suggested its applicability in biological samples.

  8. Effect of the hemolytic lectin CEL-III from Holothuroidea Cucumaria echinata on the ANS fluorescence responses in sensitive MDCK and resistant CHO cells.

    PubMed

    Oda, T; Shinmura, N; Nishioka, Y; Komatsu, N; Hatakeyama, T; Muramatsu, T

    1999-04-01

    The addition of CEL-III to sensitive MDCK cells preincubated with 8-anilino-1-naphthalenesulfonate (ANS) caused an increase in the fluorescence intensity of the probe. The increase in the ANS fluorescence caused by CEL-III was Ca2+-dependent and strongly inhibited by 0.1 M lactose, indicating that Ca2+-dependent binding of CEL-III to specific carbohydrate receptors on the plasma membrane is responsible for this phenomenon. In contrast, no significant effect of CEL-III on the ANS fluorescence was observed in CHO cells, which are highly resistant to CEL-III cytotoxicity. In MDCK cells, energy transfer from tryptophan residues to bound ANS molecules was observed in the presence of CEL-III, but not in CHO cells. Furthermore, the amount of ANS bound to MDCK cells increased as the concentration of CEL-III increased. Therefore, a simple interpretation is that the CEL-III-induced increase in ANS fluorescence is attributable to an increase of the hydrophobic region in the plasma membrane where ANS could bind. Immunoblotting analysis of proteins from cells treated with CEL-III indicated that CEL-III oligomers were irreversibly bound to the cells, and the amount of oligomer bound to MDCK cells was much greater than that bound to CHO cells under any conditions tested. The oligomerization may be accompanied by an enhancement of the hydrophobicity of CEL-III molecules, which in turn provides new ANS-binding sites. The difference in susceptibility of MDCK and CHO cells to CEL-III cytotoxicity may be due to a difference in oligomerization of bound CEL-III.

  9. A molecular imprinting-based turn-on Ratiometric fluorescence sensor for highly selective and sensitive detection of 2,4-dichlorophenoxyacetic acid (2,4-D).

    PubMed

    Wang, Xiaoyan; Yu, Jialuo; Wu, Xiaqing; Fu, Junqing; Kang, Qi; Shen, Dazhong; Li, Jinhua; Chen, Lingxin

    2016-07-15

    A novel molecular imprinting-based turn-on ratiometric fluorescence sensor was constructed via a facile sol-gel polymerization for detection of 2,4-dichlorophenoxyacetic acid (2,4-D) on the basis of photoinduced electron transfer (PET) by using nitrobenzoxadiazole (NBD) as detection signal source and quantum dots (QDs) as reference signal source. With the presence and increase of 2,4-D, the amine groups on the surface of QDs@SiO2 could bind with 2,4-D and thereby the NBD fluorescence intensities could be significantly enhanced since the PET process was inhibited, while the QDs maintained constant intensities. Accordingly, the ratio of the dual-emission intensities of green NBD and red QDs could be utilized for turn-on fluorescent detection of 2,4-D, along with continuous color changes from orange-red to green readily observed by the naked eye. The as-prepared fluorescence sensor obtained high sensitivity with a low detection limit of 0.14μM within 5min, and distinguished recognition selectivity for 2,4-D over its analogs. Moreover, the sensor was successfully applied to determine 2,4-D in real water samples, and high recoveries at three spiking levels of 2,4-D ranged from 95.0% to 110.1% with precisions below 4.5%. The simple, rapid and reliable visual sensing strategy would not only provide potential applications for high selective ultratrace analysis of complicated matrices, but also greatly enrich the research connotations of molecularly imprinted sensors.

  10. Fluorescence biosensing strategy based on mercury ion-mediated DNA conformational switch and nicking enzyme-assisted cycling amplification for highly sensitive detection of carbamate pesticide.

    PubMed

    Wang, Xiuzhong; Hou, Ting; Dong, Shanshan; Liu, Xiaojuan; Li, Feng

    2016-03-15

    Pesticides are of great importance in agricultural and biological fields, but pesticide residues may harm the environment and human health. A highly sensitive fluorescent biosensor for the detection of carbamate pesticide has been developed based on acetylcholinesterase (AChE)-catalyzed hydrolysis product triggered Hg(2+) release coupled with subsequent nicking enzyme-induced cleavage of a duplex DNA for cycling amplification. In this protocol, two DNA probes, an unmodified single-stranded helper DNA probe 1 (HP1) and a quencher-fluorophore probe (QFP) are ingeniously designed. HP1 can be folded into hairpin configuration through T-Hg(2+)-T base pair formation. QFP, labeled with FAM and BHQ1 at its two terminals, contains the recognition sequence and the cleavage site of the nicking enzyme. In the presence of carbamate pesticide, the activity of AChE is inhibited, and the amount of the product containing the thiol group generated by the hydrolysis reaction of acetylthiocholine chloride (ACh) decreases, resulting in the release of a low concentration of Hg(2+). The number of HP1 that can be selectively unfolded would be reduced and the subsequent nicking enzyme-assisted cleavage processes would be affected, resulting in decreased fluorescence signals. The fluorescence intensity further decreases with the increase of the pesticide concentration. Therefore, the pesticide content can be easily obtained by monitoring the fluorescence signal change, which is inversely proportional to the logarithm of the pesticide concentration. The detection limit of aldicarb, the model analyte, is 3.3 μgL(-1), which is much lower than the Chinese National Standards or those previously reported. The as-proposed method has also been applied to detect carbamate pesticide residues in fresh ginger and artificial lake water samples with satisfactory results, which demonstrates that the method has great potential for practical application in biological or food safety field.

  11. Label-free and sensitive fluorescence detection of nucleic acid, based on combination of a graphene oxid /SYBR green I dye platform and polymerase assisted signal amplification

    NASA Astrophysics Data System (ADS)

    Zhu, Xiao; Xing, Da

    2012-12-01

    A new label-free isothermal fluorescence amplification detection for nucleic acid has been developed. In this paper, we first developed a novel sensitive and specific detection platform with an unmodified hairpin probe (HP) combination of the graphene oxid (GO)/ SYBR green I dye (SG), which was relied on the selective principle of adsorption and the high quenching efficiency of GO. Then for the application of this new strategy, we used Mirco RNA-21 (Mir-21) as the target to evaluate this working principle of our design. When the target was hybridizing with the HP and inducing its conformation of change, an efficient isothermal circular strand-displacement polymerization reaction was activating to assist the first signal amplification. In this format, the formed complex conformation of DNA would interact with its high affinity dye, then detached from the surface of GO after incubating with the platform of GO/intercalating dye. This reaction would accompany with obvious fluorescence recovery, and accomplish farther signal enhancement by a mass of intercalating dye inserting into the minor groove of the long duplex replication product. By taking advantage of the multiple amplification of signal, this method exerted substantial enhancement in sensitivity and could be used for rapid and selective detection of Mir-21 with attomole range. It is expected that this cost-effective GO based sensor might hold considerable potential to apply in bioanalysis studies.

  12. Development of a potential method based on microchip electrophoresis with fluorescence detection for the sensitive determination of intracellular thiols in RAW264.7 cells.

    PubMed

    Tu, Feng-Qin; Zhang, Li-Yun; Guo, Xiao-Feng; Wang, Hong; Zhang, Hua-Shan

    2014-04-01

    This paper, for the first time, reported the development of a simple, rapid, and reliable method for the separation and sensitive determination of four thiol compounds including homocysteine, cysteine, glutathione, and N-acetylcysteine based on glass MCE with fluorescence detection using a highly reactive fluorogenic probe, 1,3,5,7-tetramethyl-8-phenyl-(2-maleimide)-difluoroboradiaza-s-indacene (TMPAB-o-M), as the labeling reagent. TMPAB-o-M reacted selectively with thiols to produce highly fluorescent derivatives and the highest derivatization efficiency was achieved within 6 min in physiological conditions. After the optimization of separation conditions, a baseline separation of the four thiol compounds was achieved with the detection limits ranging from 2 nM for glutathione to 4 nM for cysteine (S/N = 3) and RSDs (n = 5) in the range of 3.2-3.8%. The proposed method was significantly sensitive compared to those using electrochemical or even LIF detection in MCE-based setup reported previously, and applied to the determination of intracellular thiols in macrophage RAW264.7 cells.

  13. A Two-Step Lyssavirus Real-Time Polymerase Chain Reaction Using Degenerate Primers with Superior Sensitivity to the Fluorescent Antigen Test

    PubMed Central

    Nazé, Florence; Francart, Aurélie; Lamoral, Sophie; De Craeye, Stéphane; Kalai, Michael

    2014-01-01

    A generic two-step lyssavirus real-time reverse transcriptase polymerase chain reaction (qRT-PCR), based on a nested PCR strategy, was validated for the detection of different lyssavirus species. Primers with 17 to 30% of degenerate bases were used in both consecutive steps. The assay could accurately detect RABV, LBV, MOKV, DUVV, EBLV-1, EBLV-2, and ABLV. In silico sequence alignment showed a functional match with the remaining lyssavirus species. The diagnostic specificity was 100% and the sensitivity proved to be superior to that of the fluorescent antigen test. The limit of detection was ≤1 50% tissue culture infectious dose. The related vesicular stomatitis virus was not recognized, confirming the selectivity for lyssaviruses. The assay was applied to follow the evolution of rabies virus infection in the brain of mice from 0 to 10 days after intranasal inoculation. The obtained RNA curve corresponded well with the curves obtained by a one-step monospecific RABV-qRT-PCR, the fluorescent antigen test, and virus titration. Despite the presence of degenerate bases, the assay proved to be highly sensitive, specific, and reproducible. PMID:24822188

  14. Nitrogen and Phosphorus Co-Doped Carbon Nanodots as a Novel Fluorescent Probe for Highly Sensitive Detection of Fe(3+) in Human Serum and Living Cells.

    PubMed

    Shi, Bingfang; Su, Yubin; Zhang, Liangliang; Huang, Mengjiao; Liu, Rongjun; Zhao, Shulin

    2016-05-04

    Chemical doping with heteroatoms can effectively modulate physicochemical and photochemical properties of carbon dots (CDs). However, the development of multi heteroatoms codoped carbon nanodots is still in its early stage. In this work, a facile hydrothermal synthesis strategy was applied to synthesize multi heteroatoms (nitrogen and phosphorus) codoped carbon nanodots (N,P-CDs) using glucose as carbon source, and ammonia, phosphoric acid as dopant, respectively. Compared with CDs, the multi heteroatoms doped CDs resulted in dramatic improvement in the electronic characteristics and surface chemical activities. Therefore, the N,P-CDs prepared as described above exhibited a strong blue emission and a sensitive response to Fe(3+). The N,P-CDs based fluorescent sensor was then applied to sensitively determine Fe(3+) with a detection limit of 1.8 nM. Notably, the prepared N,P-CDs possessed negligible cytotoxicity, excellent biocompatibility, and high photostability. It was also applied for label-free detection of Fe(3+) in complex biological samples and the fluorescence imaging of intracellular Fe(3+), which indicated its potential applications in clinical diagnosis and other biologically related study.

  15. Fluorescence enhancement of CdTe quantum dots by HBcAb-HRP for sensitive detection of H2O2 in human serum.

    PubMed

    Gong, Tingting; Liu, Junfeng; Wu, Yiwei; Xiao, Yao; Wang, Xuehan; Yuan, Siqi

    2017-06-15

    A simple, high selective, ultra-sensitive and stable biosensor based on hepatitis B core antibody labeled with horseradish peroxidase (HBcAb-HRP) induced fluorescent enhancement of CdTe QDs for recognition of H2O2 have been constructed. In this assay, sulfurs in HBcAb-HRP, which possess a strong affinity towards Cd(2+), can improve greatly the recombination fluorescence of CdTe QDs by creating more radiative centers at CdTe/Cd-SR complex. Then, H2O2 oxidizes Cd-S bonds in CdTe QDs to organic disulfide product (RS-SR), causin