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Sample records for cell-based fluorescence resonance

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

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

    2011-04-01

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

  2. Cell-based lipid flippase assay employing fluorescent lipid derivatives

    Jensen, Maria Stumph; Costa, Sara; Günther-Pomorski, Thomas

    2016-01-01

    P-type ATPases in the P4 subfamily (P4-ATPases) are transmembrane proteins unique for eukaryotes that act as lipid flippases, i.e., to translocate phospholipids from the exofacial to the cytofacial monolayer of cellular membranes. While initially characterized as aminophospholipid translocases, s...... flippase activities in the plasma membrane of cells, using yeast as an example.......P-type ATPases in the P4 subfamily (P4-ATPases) are transmembrane proteins unique for eukaryotes that act as lipid flippases, i.e., to translocate phospholipids from the exofacial to the cytofacial monolayer of cellular membranes. While initially characterized as aminophospholipid translocases......, studies of individual P4-ATPase family members from fungi, plants, and animals show that P4-ATPases differ in their substrate specificities and mediate transport of a broader range of lipid substrates. Here, we describe an assay based on fluorescent lipid derivatives to monitor and characterize lipid...

  3. Magnetic resonance imaging and cell-based neurorestorative therapy after brain injury

    Quan Jiang

    2016-01-01

    Full Text Available Restorative cell-based therapies for experimental brain injury, such as stroke and traumatic brain injury, substantially improve functional outcome. We discuss and review state of the art magnetic resonance imaging methodologies and their applications related to cell-based treatment after brain injury. We focus on the potential of magnetic resonance imaging technique and its associated challenges to obtain useful new information related to cell migration, distribution, and quantitation, as well as vascular and neuronal remodeling in response to cell-based therapy after brain injury. The noninvasive nature of imaging might more readily help with translation of cell-based therapy from the laboratory to the clinic.

  4. Magnetic resonance tracking of fluorescent nanodiamond fabrication

    Shames, A. I.; Osipov, V. Yu; Boudou, J. P.; Panich, A. M.; von Bardeleben, H. J.; Treussart, F.; Vul', A. Ya

    2015-04-01

    Magnetic resonance techniques (electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR)) are used for tracking the multi-stage process of the fabrication of fluorescent nanodiamonds (NDs) produced by high-energy electron irradiation, annealing, and subsequent nano-milling. Pristine commercial high pressure and high temperature microdiamonds (MDs) with mean size 150 μm contain ~5  ×  1018 spins/g of singlet (S = 1/2) substitutional nitrogen defects P1, as well as sp3 C-C dangling bonds in the crystalline lattice. The half-field X-band EPR clearly shows (by the appearance of the intense ‘forbidden’ g = 4.26 line) that high-energy electron irradiation and annealing of MDs induce a large amount (~5  ×  1017 spins/g) of triplet (S = 1) magnetic centers, which are identified as negatively charged nitrogen vacancy defects (NV-). This is supported by EPR observations of the ‘allowed’ transitions between Zeeman sublevels of the triplet state. After progressive milling of the fluorescent MDs down to an ultrasubmicron scale (≤100 nm), the relative abundance of EPR active NV- defects in the resulting fluorescent NDs (FND) substantially decreases and, vice versa, the content of C-inherited singlet defects correlatively increases. In the fraction of the finest FNDs (mean particle size fingerprint of the presence of NV- centers in small ND systems. The same size reduction causes the disappearance of the characteristic hyperfine satellites in the spectra of the P1 centers. We discuss the mechanisms that cause both the strong reduction of the peak intensity of the ‘allowed’ lines in EPR spectra of triplet defects and the transformation of the P1 spectra.

  5. A cell-based fluorescent glucose transporter assay for SGLT2 inhibitor discovery

    Yi Huan

    2013-04-01

    Full Text Available The sodium/glucose cotransporter 2 (SGLT2 is responsible for the majority of glucose reabsorption in the kidney, and currently, SGLT2 inhibitors are considered as promising hypoglycemic agents for the treatment of type 2 diabetes mellitus. By constructing CHO cell lines that stably express the human SGLT2 transmembrane protein, along with a fluorescent glucose transporter assay that uses 2-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-ylamino]2-deoxyglucose (2-NBDG as a glucose analog, we have developed a nonradioactive, cell-based assay for the discovery and characterization of SGLT2 inhibitors.

  6. Magnetic resonance tracking of fluorescent nanodiamond fabrication

    Shames, A I; Panich, A M; Osipov, V Yu; Vul’, A Ya; Boudou, J P; Treussart, F; Von Bardeleben, H J

    2015-01-01

    Magnetic resonance techniques (electron paramagnetic resonance (EPR) and nuclear magnetic resonance (NMR)) are used for tracking the multi-stage process of the fabrication of fluorescent nanodiamonds (NDs) produced by high-energy electron irradiation, annealing, and subsequent nano-milling. Pristine commercial high pressure and high temperature microdiamonds (MDs) with mean size 150 μm contain ∼5  ×  10 18  spins/g of singlet (S = 1/2) substitutional nitrogen defects P1, as well as sp 3 C–C dangling bonds in the crystalline lattice. The half-field X-band EPR clearly shows (by the appearance of the intense ‘forbidden’ g = 4.26 line) that high-energy electron irradiation and annealing of MDs induce a large amount (∼5  ×  10 17  spins/g) of triplet (S = 1) magnetic centers, which are identified as negatively charged nitrogen vacancy defects (NV − ). This is supported by EPR observations of the ‘allowed’ transitions between Zeeman sublevels of the triplet state. After progressive milling of the fluorescent MDs down to an ultrasubmicron scale (≤100 nm), the relative abundance of EPR active NV − defects in the resulting fluorescent NDs (FND) substantially decreases and, vice versa, the content of C-inherited singlet defects correlatively increases. In the fraction of the finest FNDs (mean particle size <20 nm), which are contained in the dried supernatant of ultracentrifuged aqueous dispersion of FNDs, the NV − content is found to be reduced by one order of magnitude whereas the singlet defects content increases up to ∼2  ×  10 19  spins/g. In addition, another triplet-type defect, which is characterized by the g = 4.00 ‘forbidden’ line, appears. On reduction of the particle size below the 20 nm limit, the ‘allowed’ EPR lines become practically unobservable, whereas the ‘forbidden’ lines remain as a reliable fingerprint of the presence of NV − centers in small ND systems. The same size reduction

  7. Fluorescence Resonance Energy Transfer in Polydiacetylene Liposomes

    Li, Xuelian; Matthews, Shelton; Kohli, Punit

    2009-01-01

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

  8. Photon-number statistics in resonance fluorescence

    Lenstra, D.

    1982-12-01

    The theory of photon-number statistics in resonance fluorescence is treated, starting with the general formula for the emission probability of n photons during a given time interval T. The results fully confirm formerly obtained results by Cook that were based on the theory of atomic motion in a traveling wave. General expressions for the factorial moments are derived and explicit results for the mean and the variance are given. It is explicitly shown that the distribution function tends to a Gaussian when T becomes much larger than the natural lifetime of the excited atom. The speed of convergence towards the Gaussian is found to be typically slow, that is, the third normalized central moment (or the skewness) is proportional to T-12. However, numerical results illustrate that the overall features of the distribution function are already well represented by a Gaussian when T is larger than a few natural lifetimes only, at least if the intensity of the exciting field is not too small and its detuning is not too large.

  9. A Fluorescent Cell-Based System for Imaging Zika Virus Infection in Real-Time

    Michael J. McFadden

    2018-02-01

    Full Text Available Zika virus (ZIKV is a re-emerging flavivirus that is transmitted to humans through the bite of an infected mosquito or through sexual contact with an infected partner. ZIKV infection during pregnancy has been associated with numerous fetal abnormalities, including prenatal lethality and microcephaly. However, until recent outbreaks in the Americas, ZIKV has been relatively understudied, and therefore the biology and pathogenesis of ZIKV infection remain incompletely understood. Better methods to study ZIKV infection in live cells could enhance our understanding of the biology of ZIKV and the mechanisms by which ZIKV contributes to fetal abnormalities. To this end, we developed a fluorescent cell-based reporter system allowing for live imaging of ZIKV-infected cells. This system utilizes the protease activity of the ZIKV non-structural proteins 2B and 3 (NS2B-NS3 to specifically mark virus-infected cells. Here, we demonstrate the utility of this fluorescent reporter for identifying cells infected by ZIKV strains of two lineages. Further, we use this system to determine that apoptosis is induced in cells directly infected with ZIKV in a cell-autonomous manner. Ultimately, approaches that can directly track ZIKV-infected cells at the single cell-level have the potential to yield new insights into the host-pathogen interactions that regulate ZIKV infection and pathogenesis.

  10. FY08 Annual Report for Nuclear Resonance Fluorescence Imaging

    Warren, Glen A.; Caggiano, Joseph A.

    2009-01-06

    FY08 annual report for project the "Nuclear Resonance Fluorescence Imaging" project. Reviews accomplishments of last 3 years, including U-235 signature search, comparison of different photon sources, and examination of NRF measurements using monochromatic photon source.

  11. Nuclear Resonance Fluorescence for Safeguards Applications

    Ludewigt, Bernhard A; Quiter, Brian J; Ambers, Scott D

    2011-02-04

    In nuclear resonance fluorescence (NRF) measurements, resonances are excited by an external photon beam leading to the emission of {gamma} rays with specific energies that are characteristic of the emitting isotope. The promise of NRF as a non-destructive analysis technique (NDA) in safeguards applications lies in its potential to directly quantify a specific isotope in an assay target without the need for unfolding the combined responses of several fissile isotopes as often required by other NDA methods. The use of NRF for detection of sensitive nuclear materials and other contraband has been researched in the past. In the safeguards applications considered here one has to go beyond mere detection and precisely quantify the isotopic content, a challenge that is discussed throughout this report. Basic NRF measurement methods, instrumentation, and the analytical calculation of NRF signal strengths are described in Section 2. Well understood modeling and simulation tools are needed for assessing the potential of NRF for safeguards and for designing measurement systems. All our simulations were performed with the radiation transport code MCNPX, a code that is widely used in the safeguards community. Our initial studies showed that MCNPX grossly underestimated the elastically scattered background at backwards angles due to an incorrect treatment of Rayleigh scattering. While new, corrected calculations based on ENDF form factors showed much better agreement with experimental data for the elastic scattering of photons on an uranium target, the elastic backscatter is still not rigorously treated. Photonuclear scattering processes (nuclear Thomson, Delbruck and Giant Dipole Resonance scattering), which are expected to play an important role at higher energies, are not yet included. These missing elastic scattering contributions were studied and their importance evaluated evaluated against data found in the literature as discussed in Section 3. A transmission experiment

  12. Resonance Fluorescence from an Artificial Atom in Squeezed Vacuum

    D. M. Toyli

    2016-07-01

    Full Text Available We present an experimental realization of resonance fluorescence in squeezed vacuum. We strongly couple microwave-frequency squeezed light to a superconducting artificial atom and detect the resulting fluorescence with high resolution enabled by a broadband traveling-wave parametric amplifier. We investigate the fluorescence spectra in the weak and strong driving regimes, observing up to 3.1 dB of reduction of the fluorescence linewidth below the ordinary vacuum level and a dramatic dependence of the Mollow triplet spectrum on the relative phase of the driving and squeezed vacuum fields. Our results are in excellent agreement with predictions for spectra produced by a two-level atom in squeezed vacuum [Phys. Rev. Lett. 58, 2539 (1987], demonstrating that resonance fluorescence offers a resource-efficient means to characterize squeezing in cryogenic environments.

  13. Resonance fluorescence and electron spin in semiconductor quantum dots

    Zhao, Yong

    2009-11-18

    The work presented in this dissertation contains the first observation of spin-resolved resonance fluorescence from a single quantum dot and its application of direct measurement of electron spin dynamics. The Mollow triplet and the Mollow quintuplet, which are the hallmarks of resonance fluorescence, are presented as the non-spin-resolved and spin-resolved resonance fluorescence spectrum, respectively. The negligible laser background contribution, the near pure radiative broadened spectrum and the anti-bunching photon statistics imply the sideband photons are background-free and near transform-limited single photons. This demonstration is a promising step towards the heralded single photon generation and electron spin readout. Instead of resolving spectrum, an alternative spin-readout scheme by counting resonance fluorescence photons under moderate laser power is demonstrated. The measurements of n-shot time-resolved resonance fluorescence readout are carried out to reveal electron spin dynamics of the measurement induced back action and the spin relaxation. Hyperfine interaction and heavy-light hole mixing are identified as the relevant mechanisms for the back action and phonon-assistant spin-orbit interaction dominates the spin relaxation. After a detailed discussion on charge-spin configurations in coupled quantum dots system, the single-shot readout on electron spin are proposed. (orig.)

  14. Resonance fluorescence and electron spin in semiconductor quantum dots

    Zhao, Yong

    2009-01-01

    The work presented in this dissertation contains the first observation of spin-resolved resonance fluorescence from a single quantum dot and its application of direct measurement of electron spin dynamics. The Mollow triplet and the Mollow quintuplet, which are the hallmarks of resonance fluorescence, are presented as the non-spin-resolved and spin-resolved resonance fluorescence spectrum, respectively. The negligible laser background contribution, the near pure radiative broadened spectrum and the anti-bunching photon statistics imply the sideband photons are background-free and near transform-limited single photons. This demonstration is a promising step towards the heralded single photon generation and electron spin readout. Instead of resolving spectrum, an alternative spin-readout scheme by counting resonance fluorescence photons under moderate laser power is demonstrated. The measurements of n-shot time-resolved resonance fluorescence readout are carried out to reveal electron spin dynamics of the measurement induced back action and the spin relaxation. Hyperfine interaction and heavy-light hole mixing are identified as the relevant mechanisms for the back action and phonon-assistant spin-orbit interaction dominates the spin relaxation. After a detailed discussion on charge-spin configurations in coupled quantum dots system, the single-shot readout on electron spin are proposed. (orig.)

  15. Resonance fluorescence from an atom in a squeezed vacuum

    Carmichael, H. J.; Lane, A. S.; Walls, D. F.

    1987-06-01

    The fluorescent spectrum for a two-level atom which is damped by a squeezed vacuum shows striking differences from the spectrum for ordinary resonance fluorescence. For strong coherent driving fields the Mollow triplet depends on the relative phase of the driving field and the squeezed vacuum field. The central peak may have either subnatural linewidth or supernatural linewidth depending on this phase. The mean atomic polarization also shows a phase sensitivity.

  16. Laser resonant ionization spectroscopy and laser-induced resonant fluorescence spectra of samarium atom

    Jin, Changtai

    1995-01-01

    We have measured new high-lying levels of Sm atom by two-colour resonant photoionisation spectroscopy; we have observed the isotope shifts of Sm atom by laser-induced resonant fluorescence spectroscopy; the lifetime of eight low-lying levels of Sm atom were measured by using pulsed laser-Boxcar technique in atomic beam.

  17. Interference in the resonance fluorescence of two incoherently coupled transitions

    Kiffner, Martin; Evers, Joerg; Keitel, Christoph H.

    2006-01-01

    The fluorescence light emitted by a four-level system in J=1/2 to J=1/2 configuration driven by a monochromatic laser field and in an external magnetic field is studied. We show that the spectrum of resonance fluorescence emitted on the π transitions shows a signature of spontaneously generated interference effects. The degree of interference in the fluorescence spectrum can be controlled by means of the external magnetic field, provided that the Lande g factors of the excited and the ground state doublet are different. For a suitably chosen magnetic field strength, the relative weight of the Rayleigh line can be completely suppressed, even for low intensities of the coherent driving field. The incoherent fluorescence spectrum emitted on the π transitions exhibits a very narrow peak whose width and weight depend on the magnetic field strength. We demonstrate that the spectrum of resonance fluorescence emitted on the σ transitions shows an indirect signature of interference. A measurement of the relative peak heights in the spectrum from the σ transitions allows us to determine the branching ratio of the spontaneous decay of each excited state into the σ channel

  18. Using magnetic resonance imaging to evaluate dendritic cell-based vaccination.

    Peter M Ferguson

    Full Text Available Cancer immunotherapy with antigen-loaded dendritic cell-based vaccines can induce clinical responses in some patients, but further optimization is required to unlock the full potential of this strategy in the clinic. Optimization is dependent on being able to monitor the cellular events that take place once the dendritic cells have been injected in vivo, and to establish whether antigen-specific immune responses to the tumour have been induced. Here we describe the use of magnetic resonance imaging (MRI as a simple, non-invasive approach to evaluate vaccine success. By loading the dendritic cells with highly magnetic iron nanoparticles it is possible to assess whether the injected cells drain to the lymph nodes. It is also possible to establish whether an antigen-specific response is initiated by assessing migration of successive rounds of antigen-loaded dendritic cells; in the face of a successfully primed cytotoxic response, the bulk of antigen-loaded cells are eradicated on-route to the node, whereas cells without antigen can reach the node unchecked. It is also possible to verify the induction of a vaccine-induced response by simply monitoring increases in draining lymph node size as a consequence of vaccine-induced lymphocyte trapping, which is an antigen-specific response that becomes more pronounced with repeated vaccination. Overall, these MRI techniques can provide useful early feedback on vaccination strategies, and could also be used in decision making to select responders from non-responders early in therapy.

  19. Imaging atoms from resonance fluorescence spectrum beyond the diffraction limit

    Liao, Zeyang; Al-Amri, Mohammad; Zubairy, M. Suhail

    2014-03-01

    We calculate the resonance fluorescence spectrum of a linear chain of two-level atoms driven by a gradient coherent laser field. The result shows that we can determine the positions of atoms from the spectrum even when the atoms locate within subwavelength range and the dipole-dipole interaction is significant. This far-field resonance fluorescence localization microscopy method does not require point-by-point scanning and it may be more time-efficient. We also give a possible scheme to extract the position information in an extended region without requiring more peak power of laser. We also briefly discuss how to do a 2D imaging based on our scheme. This work is supported by grants from the King Abdulaziz City for Science and Technology (KACST) and the Qatar National Research Fund (QNRF) under the NPRP project.

  20. Nuclear Resonance Fluorescence and Isotopic Mapping of Containers

    Johnson, Micah S.; McNabb, Dennis P.

    2009-03-01

    National security programs have expressed interest in developing systems to isotopically map shipping containers, fuel assemblies, and waste barrels for various materials including special nuclear material (SNM). Current radiographic systems offer little more than an ambiguous density silhouette of a container's contents. In this paper we will present a system being developed at LLNL to isotopically map containers using the nuclear resonance fluorescence (NRF) method. Recent experimental measurements on NRF strengths in SNM are discussed.

  1. Contraband Detection with Nuclear Resonance Fluorescence: Feasibility and Impact

    Pruet, J; Lange, D

    2007-01-01

    In this report they show that cargo interrogation systems developed to thwart trafficking of illicit nuclear materials could also be powerful tools in the larger fight against contraband smuggling. In particular, in addition to detecting special nuclear materials, cargo scanning systems that exploit nuclear resonance fluorescence to detect specific isotopes can be used to help find: chemical weapons; some drugs as well as some chemicals regulated under the controlled substances act; precious metals; materials regulated under export control laws; and commonly trafficked fluorocarbons

  2. Single-cell-based evaluation of sperm progressive motility via fluorescent assessment of mitochondria membrane potential.

    Moscatelli, Natalina; Spagnolo, Barbara; Pisanello, Marco; Lemma, Enrico Domenico; De Vittorio, Massimo; Zara, Vincenzo; Pisanello, Ferruccio; Ferramosca, Alessandra

    2017-12-20

    Sperm cells progressive motility is the most important parameter involved in the fertilization process. Sperm middle piece contains mitochondria, which play a critical role in energy production and whose proper operation ensures the reproductive success. Notably, sperm progressive motility is strictly related to mitochondrial membrane potential (MMP) and consequently to mitochondrial functionality. Although previous studies presented an evaluation of mitochondrial function through MMP assessment in entire sperm cells samples, a quantitative approach at single-cell level could provide more insights in the analysis of semen quality. Here we combine laser scanning confocal microscopy and functional fluorescent staining of mitochondrial membrane to assess MMP distribution among isolated spermatozoa. We found that the sperm fluorescence value increases as a function of growing progressive motility and that such fluorescence is influenced by MMP disruptors, potentially allowing for the discrimination of different quality classes of sperm cells in heterogeneous populations.

  3. Correlated quadratures of resonance fluorescence and the generalized uncertainty relation

    Arnoldus, Henk F.; George, Thomas F.; Gross, Rolf W. F.

    1994-01-01

    Resonance fluorescence from a two-state atom has been predicted to exhibit quadrature squeezing below the Heisenberg uncertainty limit, provided that the optical parameters (Rabi frequency, detuning, laser linewidth, etc.) are chosen carefully. When the correlation between two quadratures of the radiation field does not vanish, however, the Heisenberg limit for quantum fluctuations might be an unrealistic lower bound. A generalized uncertainty relation, due to Schroedinger, takes into account the possible correlation between the quadrature components of the radiation, and it suggests a modified definition of squeezing. We show that the coherence between the two levels of a laser-driven atom is responsible for the correlation between the quadrature components of the emitted fluorescence, and that the Schrodinger uncertainty limit increases monotonically with the coherence. On the other hand, the fluctuations in the quadrature field diminish with an increasing coherence, and can disappear completely when the coherence reaches 1/2, provided that certain phase relations hold.

  4. Resonance fluorescence based two- and three-dimensional atom localization

    Wahab, Abdul; Rahmatullah; Qamar, Sajid

    2016-06-01

    Two- and three-dimensional atom localization in a two-level atom-field system via resonance fluorescence is suggested. For the two-dimensional localization, the atom interacts with two orthogonal standing-wave fields, whereas for the three-dimensional atom localization, the atom interacts with three orthogonal standing-wave fields. The effect of the detuning and phase shifts associated with the corresponding standing-wave fields is investigated. A precision enhancement in position measurement of the single atom can be noticed via the control of the detuning and phase shifts.

  5. Performance of a Cyanobacteria Whole Cell-Based Fluorescence Biosensor for Heavy Metal and Pesticide Detection

    Salmijah Surif

    2013-05-01

    Full Text Available Whole cell biosensors always face the challenge of low stability of biological components and short storage life. This paper reports the effects of poly(2-hydroxyethyl methacrylate (pHEMA immobilization on a whole cell fluorescence biosensor for the detection of heavy metals (Cu, Pb, Cd, and pesticides (dichlorophenoxyacetic acid (2,4-D, and chlorpyrifos. The biosensor was produced by entrapping the cyanobacterium Anabaena torulosa on a cellulose membrane, followed by applying a layer of pHEMA, and attaching it to a well. The well was then fixed to an optical probe which was connected to a fluorescence spectrophotometer and an electronic reader. The optimization of the biosensor using several factors such as amount of HEMA and drying temperature were undertaken. The detection limits of biosensor without pHEMA for Cu, Cd, Pb, 2,4-D and chlorpyrifos were 1.195, 0.027, 0.0100, 0.025 and 0.025 µg/L respectively. The presence of pHEMA increased the limits of detection to 1.410, 0.250, 0.500, 0.235 and 0.117 µg/L respectively. pHEMA is known to enhance the reproducibility of the biosensor with average relative standard deviation (RSD of ±1.76% for all the pollutants tested, 48% better than the biosensor without pHEMA (RSD = ±3.73%. In storability test with Cu 5 µg/L, the biosensor with pHEMA performed 11.5% better than the test without pHEMA on day-10 and 5.2% better on day-25. pHEMA is therefore a good candidate to be used in whole cell biosensors as it increases reproducibility and enhances biosensor storability.

  6. Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications.

    Cui, Chenghua; Shu, Wei; Li, Peining

    2016-01-01

    Fluorescence in situ hybridization (FISH) is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes, and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains, and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbias and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH) allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH) using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells DNA and RNA FISH

  7. Fluorescence In situ Hybridization: Cell-Based Genetic Diagnostic and Research Applications

    Chenghua Cui

    2016-09-01

    Full Text Available Fluorescence in situ hybridization (FISH is a macromolecule recognition technology based on the complementary nature of DNA or DNA/RNA double strands. Selected DNA strands incorporated with fluorophore-coupled nucleotides can be used as probes to hybridize onto the complementary sequences in tested cells and tissues and then visualized through a fluorescence microscope or an imaging system. This technology was initially developed as a physical mapping tool to delineate genes within chromosomes. Its high analytical resolution to a single gene level and high sensitivity and specificity enabled an immediate application for genetic diagnosis of constitutional common aneuploidies, microdeletion/microduplication syndromes and subtelomeric rearrangements. FISH tests using panels of gene-specific probes for somatic recurrent losses, gains and translocations have been routinely applied for hematologic and solid tumors and are one of the fastest-growing areas in cancer diagnosis. FISH has also been used to detect infectious microbials and parasites like malaria in human blood cells. Recent advances in FISH technology involve various methods for improving probe labeling efficiency and the use of super resolution imaging systems for direct visualization of intra-nuclear chromosomal organization and profiling of RNA transcription in single cells. Cas9-mediated FISH (CASFISH allowed in situ labeling of repetitive sequences and single-copy sequences without the disruption of nuclear genomic organization in fixed or living cells. Using oligopaint-FISH and super-resolution imaging enabled in situ visualization of chromosome haplotypes from differentially specified single-nucleotide polymorphism loci. Single molecule RNA FISH (smRNA-FISH using combinatorial labeling or sequential barcoding by multiple round of hybridization were applied to measure mRNA expression of multiple genes within single cells. Research applications of these single molecule single cells

  8. A Label-Free Microfluidic Biosensor for Activity Detection of Single Microalgae Cells Based on Chlorophyll Fluorescence

    Junsheng Wang

    2013-11-01

    Full Text Available Detection of living microalgae cells is very important for ballast water treatment and analysis. Chlorophyll fluorescence is an indicator of photosynthetic activity and hence the living status of plant cells. In this paper, we developed a novel microfluidic biosensor system that can quickly and accurately detect the viability of single microalgae cells based on chlorophyll fluorescence. The system is composed of a laser diode as an excitation light source, a photodiode detector, a signal analysis circuit, and a microfluidic chip as a microalgae cell transportation platform. To demonstrate the utility of this system, six different living and dead algae samples (Karenia mikimotoi Hansen, Chlorella vulgaris, Nitzschia closterium, Platymonas subcordiformis, Pyramidomonas delicatula and Dunaliella salina were tested. The developed biosensor can distinguish clearly between the living microalgae cells and the dead microalgae cells. The smallest microalgae cells that can be detected by using this biosensor are 3 μm ones. Even smaller microalgae cells could be detected by increasing the excitation light power. The developed microfluidic biosensor has great potential for in situ ballast water analysis.

  9. A Label-Free Microfluidic Biosensor for Activity Detection of Single Microalgae Cells Based on Chlorophyll Fluorescence

    Wang, Junsheng; Sun, Jinyang; Song, Yongxin; Xu, Yongyi; Pan, Xinxiang; Sun, Yeqing; Li, Dongqing

    2013-01-01

    Detection of living microalgae cells is very important for ballast water treatment and analysis. Chlorophyll fluorescence is an indicator of photosynthetic activity and hence the living status of plant cells. In this paper, we developed a novel microfluidic biosensor system that can quickly and accurately detect the viability of single microalgae cells based on chlorophyll fluorescence. The system is composed of a laser diode as an excitation light source, a photodiode detector, a signal analysis circuit, and a microfluidic chip as a microalgae cell transportation platform. To demonstrate the utility of this system, six different living and dead algae samples (Karenia mikimotoi Hansen, Chlorella vulgaris, Nitzschia closterium, Platymonas subcordiformis, Pyramidomonas delicatula and Dunaliella salina) were tested. The developed biosensor can distinguish clearly between the living microalgae cells and the dead microalgae cells. The smallest microalgae cells that can be detected by using this biosensor are 3 μm ones. Even smaller microalgae cells could be detected by increasing the excitation light power. The developed microfluidic biosensor has great potential for in situ ballast water analysis. PMID:24287532

  10. Nuclear resonance fluorescence of {sup 203,205}Tl

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

    2009-07-01

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

  11. Fluorescence-enhanced gadolinium-doped zinc oxide quantum dots for magnetic resonance and fluorescence imaging.

    Liu, Yanlan; Ai, Kelong; Yuan, Qinghai; Lu, Lehui

    2011-02-01

    We report here the development of Gd-doped ZnO quantum dots (QDs) as dual modal fluorescence and magnetic resonance imaging nanoprobes. They are fabricated in a simple, versatile and environmentally friendly method, not only decreasing the difficulty and complexity, but also avoiding the increase of particle's size brought about by silica coating procedure in the synthesis of nanoprobes reported previously. These nanoprobes, with exceptionally small size and enhanced fluorescence resulting from the Gd doping, can label successfully the HeLa cells in short time and present no evidence of toxicity or adverse affect on cell growth even at the concentration up to 1 mm. These results show that such nanoprobes have low toxicity, especially in comparison with the traditional PEGylated CdSe/ZnS or CdSe/CdS QDs. In MRI studies, they exert strong positive contrast effect with a large longitudinal relaxivity (r(1)) of water proton of 16 mm(-1) s(-1). Their capability of imaging HeLa cells with MRI implies that they have great potential as MRI contrast agents. Combining the high sensitivity of fluorescence imaging with high spatial resolution of MRI, We expect that the as-prepared Gd-doped Zno QDs can provide a better reliability of the collected data and find promising applications in biological, medical and other fields. Copyright © 2010 Elsevier Ltd. All rights reserved.

  12. Resonance fluorescence microscopy via three-dimensional atom localization

    Panchadhyayee, Pradipta; Dutta, Bibhas Kumar; Das, Nityananda; Mahapatra, Prasanta Kumar

    2018-02-01

    A scheme is proposed to realize three-dimensional (3D) atom localization in a driven two-level atomic system via resonance fluorescence. The field arrangement for the atom localization involves the application of three mutually orthogonal standing-wave fields and an additional traveling-wave coupling field. We have shown the efficacy of such field arrangement in tuning the spatially modulated resonance in all directions. Under different parametric conditions, the 3D localization patterns originate with various shapes such as sphere, sheets, disk, bowling pin, snake flute, flower vase. High-precision localization is achieved when the radiation field detuning equals twice the combined Rabi frequencies of the standing-wave fields. Application of a traveling-wave field of suitable amplitude at optimum radiation field detuning under symmetric standing-wave configuration leads to 100% detection probability even in sub-wavelength domain. Asymmetric field configuration is also taken into consideration to exhibit atom localization with appreciable precision compared to that of the symmetric case. The momentum distribution of the localized atoms is found to follow the Heisenberg uncertainty principle under the validity of Raman-Nath approximation. The proposed field configuration is suitable for application in the study of atom localization in an optical lattice arrangement.

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

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

    2007-01-01

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

  14. [Fluorescence Resonance Energy Transfer Detection of Cobalt Ions by Silver Triangular Nanoplates and Rhodamine 6G].

    Zhang, Xiu-qing; Peng, Jun; Ling, Jian; Liu, Chao-juan; Cao, Qiu-e; Ding, Zhong-tao

    2015-04-01

    In the present paper, the authors studied fluorescence resonance energy transfer (FRET) phenomenon between silver triangular nanoplates and bovine serum albumin (BSA)/Rhodamine 6G fluorescence complex, and established a fluorescence method for the detection of cobalt ions. We found that when increasing the silver triangular nanoplates added to certain concentrations of fluorescent bovine serum albumin (BSA)/Rhodamine 6G complex, the fluorescence of Rhodamine 6G would be quenched up to 80% due to the FRET between the quencher and donor. However, in the presence of cobalt ions, the disassociation of the fluorescent complex from silver triangular nanoplates occurred and the fluorescence of the Rhodamine 6G recovered. The recovery of fluorescence intensity rate (I/I0) has a good relationship with the cobalt ion concentration (cCO2+) added. Thus, the authors developed a fluorescence method for the detection of cobalt ions based on the FRET of silver triangular nanoplates and Rhodamine 6G.

  15. Development of cell-based quantitative evaluation method for cell cycle-arrest type cancer drugs for apoptosis by high precision surface plasmon resonance sensor

    Ona, Toshihiro; Nishijima, Hiroshi; Kosaihira, Atsushi; Shibata, Junko

    2008-04-01

    In vitro rapid and quantitative cell-based assay is demanded to verify the efficacy prediction of cancer drugs since a cancer patient may have unconventional aspects of tumor development. Here, we show the rapid and non-label quantitative verifying method and instrumentation of apoptosis for cell cycle-arrest type cancer drugs (Roscovitine and D-allose) by reaction analysis of living liver cancer cells cultured on a sensor chip with a newly developed high precision (50 ndeg s -1 average fluctuation) surface plasmon resonance (SPR) sensor. The time-course cell reaction as the SPR angle change rate for 10 min from 30 min cell culture with a drug was significantly related to cell viability. By the simultaneous detection of differential SPR angle change and fluorescence by specific probes using the new instrument, the SPR angle was related to the nano-order potential decrease in inner mitochondrial membrane potential. The results obtained are universally valid for the cell cycle-arrest type cancer drugs, which mediate apoptosis through different cell-signaling pathways, by a liver cancer cell line of Hep G2 (P<0.001). This system towards the application to evaluate personal therapeutic potentials of drugs using cancer cells from patients in clinical use.

  16. Resonance fluorescence spectra of a three-level atom driven by two strong laser fields

    Peng Jinsheng.

    1986-12-01

    The resonance fluorescence of a three-level atom interacted with two high-power laser fields is investigated in strong field approximation. The fluorescence distribution is obtained by means of the theory of dressing transformation. (author). 15 refs, 2 figs

  17. Near-infrared emitting fluorescent nanocrystals-labeled natural killer cells as a platform technology for the optical imaging of immunotherapeutic cells-based cancer therapy

    Lim, Yong Taik; Cho, Mi Young; Noh, Young-Woock; Chung, Bong Hyun; Chung, Jin Woong

    2009-01-01

    This study describes the development of near-infrared optical imaging technology for the monitoring of immunotherapeutic cell-based cancer therapy using natural killer (NK) cells labeled with fluorescent nanocrystals. Although NK cell-based immunotherapeutic strategies have drawn interest as potent preclinical or clinical methods of cancer therapy, there are few reports documenting the molecular imaging of NK cell-based cancer therapy, primarily due to the difficulty of labeling of NK cells with imaging probes. Human natural killer cells (NK92MI) were labeled with anti-human CD56 antibody-coated quantum dots (QD705) for fluorescence imaging. FACS analysis showed that the NK92MI cells labeled with anti-human CD56 antibody-coated QD705 have no effect on the cell viability. The effect of anti-human CD56 antibody-coated QD705 labeling on the NK92MI cell function was investigated by measuring interferon gamma (IFN- γ) production and cytolytic activity. Finally, the NK92MI cells labeled with anti-human CD56 antibody-coated QD705 showed a therapeutic effect similar to that of unlabeled NK92MI cells. Images of intratumorally injected NK92MI cells labeled with anti-human CD56 antibody-coated could be acquired using near-infrared optical imaging both in vivo and in vitro. This result demonstrates that the immunotherapeutic cells labeled with fluorescent nanocrystals can be a versatile platform for the effective tracking of injected therapeutic cells using optical imaging technology, which is very important in cell-based cancer therapies.

  18. Nuclear Resonance Fluorescence off 54Cr: The Onset of the Pygmy Dipole Resonance

    Ries, P. C.; Beck, T.; Beller, J.; Krishichayan; Gayer, U.; Isaak, J.; Löher, B.; Mertes, L.; Pai, H.; Pietralla, N.; Romig, C.; Savran, D.; Schilling, M.; Tornow, W.; Werner, V.; Zweidinger, M.

    2016-06-01

    Low-lying electric and magnetic dipole excitations (E1 and M1) below the neutron separation threshold, particularly the Pygmy Dipole Resonance (PDR), have drawn considerable attention in the last years. So far, mostly moderately heavy nuclei in the mass regions around A = 90 and A = 140 were examined with respect to the PDR. In the present work, the systematics of the PDR have been extended by measuring excitation strengths and parity quantum numbers of J = 1 states in lighter nuclei near A = 50 in order to gather information on the onset of the PDR. The nuclei 50,52,54Cr and 48,50Ti were examined via bremsstrahlung produced at the DArmstadt Superconducting electron Linear Accelerator (S-DALINAC) with photon energies up to 9.7 MeV with the method of nuclear resonance fluorescence. Numerous excited states were observed, many of which for the first time. The parity quantum numbers of these states have been determined at the High Intensity Gamma-ray Source (HIγS) of the Triangle Universities Nuclear Laboratory in Durham, NC, USA. Informations to the methods and the experimental setups will be provided and the results on 54Cr achieved will be discussed with respect to the onset of the PDR.

  19. Enhanced escape rate for Hg 254 nm resonance radiation in fluorescent lamps

    Lawler, James E; Raizen, Mark G

    2013-01-01

    The potential of the low-cost MAGIS isotopic separation method to improve fluorescent lamp efficacy is explored using resonance radiation transport simulations. New Hg isotopic mixes are discovered that yield escape rates for 254 nm Hg I resonance radiation equal to 117% to 122% of the rate for a natural isotopic mix under the same lamp conditions. (paper)

  20. Preparation and characterization of alginate based-fluorescent magnetic nanoparticles for fluorescence/magnetic resonance multimodal imaging applications

    Kwon, Yong-Su; Choi, Kee-Bong; Lim, Hyungjun; Lee, Sunghwi; Lee, Jae-Jong

    2018-06-01

    Simple and versatile methodologies have been reported that customize the surface of superparamagnetic iron oxide (SPIO) nanoparticles and impart additional fluorescence capabilities to these contrast agents. Herein, we present the rational design, synthesis, characterization, and biological applications of a new magnetic-based fluorescent probe. The dual modality imaging protocol was developed by labeling fluorophore with alginate natural polymers that have excellent biocompatibility and biodegradability, and using gelification method to form nanocomposites containing SPIO. The formation of alginate-based fluorescent magnetic (AFM) nanoparticles was observed in spherical and elliptical forms with a diameter of less than 500 nm by a transmission electron microscope (TEM). The fluorescent wavelength band in the range of 560 nm was also confirmed in the UV–visible spectrophotometer. In this study, we demonstrate that the multi-tasking design of AFM nanoparticles provides an ideal platform for building balanced dual-image probes of magnetic resonance imaging and optical imaging.

  1. Experimental models of brain ischemia: a review of techniques, magnetic resonance imaging and investigational cell-based therapies

    Alessandra eCanazza

    2014-02-01

    Full Text Available Stroke continues to be a significant cause of death and disability worldwide. Although major advances have been made in the past decades in prevention, treatment and rehabilitation, enormous challenges remain in the way of translating new therapeutic approaches from bench to bedside. Thrombolysis, while routinely used for ischemic stroke, is only a viable option within a narrow time window. Recently, progress in stem cell biology has opened up avenues to therapeutic strategies aimed at supporting and replacing neural cells in infarcted areas. Realistic experimental animal models are crucial to understand the mechanisms of neuronal survival following ischemic brain injury and to develop therapeutic interventions. Current studies on experimental stroke therapies evaluate the efficiency of neuroprotective agents and cell-based approaches using primarily rodent models of permanent or transient focal cerebral ischemia. In parallel, advancements in imaging techniques permit better mapping of the spatial-temporal evolution of the lesioned cortex and its functional responses. This review provides a condensed conceptual review of the state of the art of this field, from models and magnetic resonance imaging techniques through to stem cell therapies.

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

    Nahar, Sultana N.; Pradhan, Anil K.

    2015-04-01

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

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

    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).

  4. A BIOSENSOR USING COUPLED PLASMON WAVEGUIDE RESONANCE COMBINED WITH HYPERSPECTRAL FLUORESCENCE ANALYSIS

    CHAN DU

    2014-01-01

    Full Text Available We developed a biosensor that is capable for simultaneous surface plasmon resonance (SPR sensing and hyperspectral fluorescence analysis in this paper. A symmetrical metal-dielectric slab scheme is employed for the excitation of coupled plasmon waveguide resonance (CPWR in the present work. Resonance between surface plasmon mode and the guided waveguide mode generates narrower full width half-maximum of the reflective curves which leads to increased precision for the determination of refractive index over conventional SPR sensors. In addition, CPWR also offers longer surface propagation depths and higher surface electric field strengths that enable the excitation of fluorescence with hyperspectral technique to maintain an appreciable signal-to-noise ratio. The refractive index information obtained from SPR sensing and the chemical properties obtained through hyperspectral fluorescence analysis confirm each other to exclude false-positive or false-negative cases. The sensor provides a comprehensive understanding of the biological events on the sensor chips.

  5. Metal Nanoparticles/Porous Silicon Microcavity Enhanced Surface Plasmon Resonance Fluorescence for the Detection of DNA

    Jiajia Wang

    2018-02-01

    Full Text Available A porous silicon microcavity (PSiMC with resonant peak wavelength of 635 nm was fabricated by electrochemical etching. Metal nanoparticles (NPs/PSiMC enhanced fluorescence substrates were prepared by the electrostatic adherence of Au NPs that were distributed in PSiMC. The Au NPs/PSiMC device was used to characterize the target DNA immobilization and hybridization with its complementary DNA sequences marked with Rhodamine red (RRA. Fluorescence enhancement was observed on the Au NPs/PSiMC device substrate; and the minimum detection concentration of DNA ran up to 10 pM. The surface plasmon resonance (SPR of the MC substrate; which is so well-positioned to improve fluorescence enhancement rather the fluorescence enhancement of the high reflection band of the Bragg reflector; would welcome such a highly sensitive in biosensor.

  6. Metal Nanoparticles/Porous Silicon Microcavity Enhanced Surface Plasmon Resonance Fluorescence for the Detection of DNA.

    Wang, Jiajia; Jia, Zhenhong

    2018-02-23

    A porous silicon microcavity (PSiMC) with resonant peak wavelength of 635 nm was fabricated by electrochemical etching. Metal nanoparticles (NPs)/PSiMC enhanced fluorescence substrates were prepared by the electrostatic adherence of Au NPs that were distributed in PSiMC. The Au NPs/PSiMC device was used to characterize the target DNA immobilization and hybridization with its complementary DNA sequences marked with Rhodamine red (RRA). Fluorescence enhancement was observed on the Au NPs/PSiMC device substrate; and the minimum detection concentration of DNA ran up to 10 pM. The surface plasmon resonance (SPR) of the MC substrate; which is so well-positioned to improve fluorescence enhancement rather the fluorescence enhancement of the high reflection band of the Bragg reflector; would welcome such a highly sensitive in biosensor.

  7. A fluorescence resonance energy transfer-based method for histone methyltransferases

    Devkota, Kanchan; Lohse, Brian; Nyby Jakobsen, Camilla

    2015-01-01

    A simple dye–quencher fluorescence resonance energy transfer (FRET)-based assay for methyltransferases was developed and used to determine kinetic parameters and inhibitory activity at EHMT1 and EHMT2. Peptides mimicking the truncated histone H3 tail were functionalized in each end with a dye...

  8. Time-resolved resonance fluorescence spectroscopy for study of chemical reactions in laser-induced plasmas.

    Liu, Lei; Deng, Leimin; Fan, Lisha; Huang, Xi; Lu, Yao; Shen, Xiaokang; Jiang, Lan; Silvain, Jean-François; Lu, Yongfeng

    2017-10-30

    Identification of chemical intermediates and study of chemical reaction pathways and mechanisms in laser-induced plasmas are important for laser-ablated applications. Laser-induced breakdown spectroscopy (LIBS), as a promising spectroscopic technique, is efficient for elemental analyses but can only provide limited information about chemical products in laser-induced plasmas. In this work, time-resolved resonance fluorescence spectroscopy was studied as a promising tool for the study of chemical reactions in laser-induced plasmas. Resonance fluorescence excitation of diatomic aluminum monoxide (AlO) and triatomic dialuminum monoxide (Al 2 O) was used to identify these chemical intermediates. Time-resolved fluorescence spectra of AlO and Al 2 O were used to observe the temporal evolution in laser-induced Al plasmas and to study their formation in the Al-O 2 chemistry in air.

  9. Spectrophotometry of Bowen resonance fluorescence lines in three planetary nebulae

    O'Dell, C. R.; Miller, Christopher O.

    1992-01-01

    The results are presented of a uniquely complete, carefully reduced set of observations of the O III Bowen fluorescence lines in the planetary nebulae NGC 6210, NGC 7027, and NGC 7662. A detailed comparison with the predictions of radiative excitation verify that some secondary lines are enhanced by selective population by the charge exchange mechanism involving O IV. Charge exchange is most important in NGC 6210, which is of significantly lower ionization than the other nebulae. In addition to the principal Bowen lines arising from Ly-alpha pumping of the O III O1 line, lines arising from pumping of the O3 line are also observed. Comparison of lines produced by O1 and O3 with the theoretical predictions of Neufeld indicate poor agreement; comparison with the theoretical predictions of Harrington show agreement with NGC 7027 and NGC 7662.

  10. Resonance fluorescence revival in a voltage-controlled semiconductor quantum dot

    Reigue, Antoine; Lemaître, Aristide; Gomez Carbonell, Carmen; Ulysse, Christian; Merghem, Kamel; Guilet, Stéphane; Hostein, Richard; Voliotis, Valia

    2018-02-01

    We demonstrate systematic resonance fluorescence recovery with near-unity emission efficiency in single quantum dots embedded in a charge-tunable device in a wave-guiding geometry. The quantum dot charge state is controlled by a gate voltage, through carrier tunneling from a close-lying Fermi sea, stabilizing the resonantly photocreated electron-hole pair. The electric field cancels out the charging/discharging mechanisms from nearby traps toward the quantum dots, responsible for the usually observed inhibition of the resonant fluorescence. Fourier transform spectroscopy as a function of the applied voltage shows a strong increase in the coherence time though not reaching the radiative limit. These charge controlled quantum dots can act as quasi-perfect deterministic single-photon emitters, with one laser pulse converted into one emitted single photon.

  11. Defining a turnover index for the correlation of biomaterial degradation and cell based extracellular matrix synthesis using fluorescent tagging techniques.

    Bardsley, Katie; Wimpenny, Ian; Wechsler, Roni; Shachaf, Yonatan; Yang, Ying; El Haj, Alicia J

    2016-11-01

    Non-destructive protocols which can define a biomaterial's degradation and its associated ability to support proliferation and/or promote extracellular matrix deposition will be an essential in vitro tool. In this study we investigate fluorescently tagged biomaterials, with varying rates of degradation and their ability to support cell proliferation and osteogenic differentiation. Changes in fluorescence of the biomaterials and the release of fluorescent soluble by-products were confirmed as accurate methods to quantify degradation. It was demonstrated that increasing rates of the selected biomaterials' degradation led to a decrease in cell proliferation and concurrently an increase in osteogenic matrix production. A novel turnover index (TI), which directly describes the effect of degradation of a biomaterial on cell behaviour, was calculated. Lower TIs for proliferation and high TIs for osteogenic marker production were observed on faster degrading biomaterials, indicating that these biomaterials supported an upregulation of osteogenic markers. This TI was further validated using an ex vivo chick femur model, where the faster degrading biomaterial, fibrin, led to an increased TI for mineralisation within an epiphyseal defect. This in vitro tool, TI, for monitoring the effect of biomaterial degradation on extracellular matrix production may well act as predictor of the selected biomaterials' performance during in vivo studies. This paper outlines a novel metric, Turnover Index (TI), which can be utilised in tissue-engineering for the comparison of a range of biomaterials. The metric sets out to define the relationship between the rate of degradation of biomaterials with the rate of cell proliferation and ECM synthesis, ultimately allowing us to tailor material for set clinical requirements. We have discovered some novel comparative findings that cells cultured on biomaterials with increased rates of degradation have lower rates of proliferation but alternatively

  12. Resonance fluorescence and quantum jumps in single atoms: Testing the randomness of quantum mechanics

    Erber, T.; Hammerling, P.; Hockney, G.; Porrati, M.; Putterman, S.; La Jolla Institute, La Jolla, California 92037; Department of Physics, University of California, Los Angeles, California 90024)

    1989-01-01

    When a single trapped 198 Hg + ion is illuminated by two lasers, each tuned to an approximate transition, the resulting fluorescence switches on and off in a series of pulses resembling a bistable telegraph. This intermittent fluorescence can also be obtained by optical pumping with a single laser. Quantum jumps between successive atomic levels may be traced directly with multiple-resonance fluorescence. Atomic transition rates and photon antibunching distributions can be inferred from the pulse statistics and compared with quantum theory. Stochastic tests also indicate that the quantum telegraphs are good random number generators. During periods when the fluorescence is switched off, the radiationless atomic currents that generate the telegraph signals can be adjusted by varying the laser illumination: if this coherent evolution of the wave functions is sustained over sufficiently long time intervals, novel interactive precision measurements, near the limits of the time-energy uncertainty relations, are possible. Copyright 1989 Academic Press, Inc

  13. Resonance fluorescence and quantum interference of a single NV center

    Ma, Yong-Hong; Zhang, Xue-Feng; Wu, E.

    2017-11-01

    The detection of a single nitrogen-vacancy center in diamond has attracted much interest, since it is expected to lead to innovative applications in various domains of quantum information, including quantum metrology, information processing and communications, as well as in various nanotechnologies, such as biological and subdiffraction limit imaging, and tests of entanglement in quantum mechanics. We propose a novel scheme of a single NV center coupled with a multi-mode superconducting microwave cavity driven by coherent fields in squeezed vacuum. We numerically investigate the spectra in-phase quadrature and out-of-phase quadrature for different driving regimes with or without detunings. It shows that the maximum squeezing can be obtained for optimal Rabi fields. Moreover, with the same parameters, the maximum squeezing is greatly increased when the detunings are nonzero compared to the resonance case.

  14. Homogeneous non-competitive bioaffinity assay based on fluorescence resonance energy transfer

    Kokko, Tiina; Kokko, Leena; Soukka, Tero; Loevgren, Timo

    2007-01-01

    A homogeneous non-competitive assay principle for measurement of small analytes based on quenching of fluorescence is described. Fluorescence resonance energy transfer (FRET) occurs between the donor, intrinsically fluorescent europium(III)-chelate conjugated to streptavidin, and the acceptor, quencher dye conjugated to biotin derivative when the biotin-quencher is bound to Eu-streptavidin. Fluorescence can be measured only from those streptavidins that are bound to biotin of the sample, while the fluorescence of the streptavidins that are not occupied by biotin are quenched by quencher-biotin conjugates. The quenching efficiencies of the non-fluorescent quencher dyes were over 95% and one dye molecule was able to quench the fluorescence of more than one europium(III)-chelate. This, however, together with the quadrovalent nature of streptavidin limited the measurable range of the assay to 0.2-2 nmol L -1 . In this study we demonstrated that FRET could be used to design a non-competitive homogeneous assay for a small analyte resulting in equal performance with competitive heterogeneous assay

  15. In situ detection of atomic and molecular iodine using Resonance and Off-Resonance Fluorescence by Lamp Excitation: ROFLEX

    J. C. Gómez Martín

    2011-01-01

    Full Text Available We demonstrate a new instrument for in situ detection of atmospheric iodine atoms and molecules based on atomic and molecular resonance and off-resonance ultraviolet fluorescence excited by lamp emission. The instrument combines the robustness, light weight, low power consumption and efficient excitation of radio-frequency discharge light sources with the high sensitivity of the photon counting technique. Calibration of I2 fluorescence is achieved via quantitative detection of the molecule by Incoherent Broad Band Cavity-enhanced Absorption Spectroscopy. Atomic iodine fluorescence signal is calibrated by controlled broad band photolysis of known I2 concentrations in the visible spectral range at atmospheric pressure. The instrument has been optimised in laboratory experiments to reach detection limits of 1.2 pptv for I atoms and 13 pptv for I2, for S/N = 1 and 10 min of integration time. The ROFLEX system has been deployed in a field campaign in northern Spain, representing the first concurrent observation of ambient mixing ratios of iodine atoms and molecules in the 1–350 pptv range.

  16. Highly Sensitive Fluorescent Sensor for Cartap Based on Fluorescence Resonance Energy Transfer Between Gold Nanoparticles and Rhodamine B.

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

    2018-04-01

    Cartap 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 ultrasensitive analytical method for the determination of cartap based on fluorescence resonance energy transfer (FRET) between Au nanoparticles (AuNPs) and rhodamine B (RB) is first described. With the presence of citrate-stabilized AuNPs, the fluorescence of RB was remarkably quenched by AuNPs via FRET. The fluorescence of the AuNPs-RB system was recovered upon addition of cartap, cartap can be adsorbed on the surface of AuNPs due to its amino group that has good affinity with gold, which could induce the aggregation of AuNPs accompanying color change from red to blue. Thus, the FRET between AuNPs and RB was weakened and the PL intensity of RB was recovered accordingly. A good linear correlation for detection of RB was exhibited from 1 nM to 180 nM, and the detection limit reached 0.88 nM, which was much lower than the safety limit required by USA, UK and China. To the best of our knowledge, it has been the lowest detection ever without the aid of costly instrumentation. This method was successfully carried out for the assessment of cartap in real samples with satisfactory results, which revealed many advantages such as high sensitivity, low cost and non-time-consuming compared with traditional methods.

  17. Advances in Spiropyrans/Spirooxazines and Applications Based on Fluorescence Resonance Energy Transfer (FRET with Fluorescent Materials

    Hongyan Xia

    2017-12-01

    Full Text Available Studies on the following were reviewed: (1 the structure of spiropyrans and spirooxazines (two kinds of spiro compounds under external stimuli and (2 the construction and applications of composite systems based on fluorescence resonance energy transfer (FRET with fluorescent materials. When treated with different stimuli (light, acids and bases, solvents, metal ions, temperature, redox potential, and so on, spiropyrans/spirooxazines undergo transformations between the ring-closed form (SP, the ring-opened merocyanine (MC form, and the protonated ring-opened form (MCH. This is due to the breakage of the spiro C–O bond and the protonation of MC, along with a color change. Various novel, multifunctional materials based on photochromic spiropyrans and spirooxazines have been successfully developed because of the vastly differently physiochemical properties posssed by the SP, MC and MCH forms. Among the three different structural forms, the MC form has been studied most extensively. The MC form not only gives complexes with various inorganic particles, biological molecules, and organic chemicals but also acts as the energy acceptor (of energy from fluorescent molecules during energy transfer processes that take place under proper conditions. Furthermore, spiropyran and spirooxazine compounds exhibit reversible physicochemical property changes under proper stimuli; this provides more advantages compared with other photochromic compounds. Additionally, the molecular structures of spiropyrans and spirooxazines can be easily modified and extended, so better compounds can be obtained to expand the scope of already known applications. Described in detail are: (1 the structural properties of spiropyrans and spirooxazines and related photochromic mechanisms; (2 composite systems based on spiropyrans and spirooxazines, and (3 fluorescent materials which have potential applications in sensing, probing, and a variety of optical elements.

  18. Far-Red Fluorescent Probe for Imaging of Vicinal Dithiol-Containing Proteins in Living Cells Based on a pKa Shift Mechanism.

    Zhang, Shengrui; Chen, Guojun; Wang, Yuanyuan; Wang, Qin; Zhong, Yaogang; Yang, Xiao-Feng; Li, Zheng; Li, Hua

    2018-02-20

    Vicinal dithiol-containing proteins (VDPs) play fundamental roles in intracellular redox homeostasis and are responsible for many diseases. In this work, we report a far-red fluorescence turn-on probe MCAs for VDPs exploiting the pK a shift of the imine functionality of the probe. MCAs is composed of a merocyanine Schiff base as the fluorescent reporter and a cyclic 1,3,2-dithiarsenolane as the specific ligand for VDPs. The imine pK a of MCAs is 4.8, and it exists predominantly in the Schiff base (SB) form at physiological pH. Due to the absence of a resonating positive charge, it absorbs at a relatively short wavelength and is essentially nonfluorescent. Upon selective binding to reduced bovine serum albumin (rBSA, selected as the model protein), MCAs was brought from aqueous media to the binding pockets of the protein, causing a large increase in pK a value of MCAs (pK a = 7.1). As a result, an increase in the protonated Schiff base (PSB) form of MCAs was observed at the physiological pH conditions, which in turn leads to a bathochromically shifted chromophore (λ abs = 634 nm) and a significant increase in fluorescence intensity (λ em = 657 nm) simultaneously. Furthermore, molecular dynamics simulations indicate that the salt bridges formed between the iminium in MCAs and the residues D72 and D517 in rBSA resist the dissociation of proton from the probe, thus inducing an increase of the pK a value. The proposed probe shows excellent sensitivity and specificity toward VDPs over other proteins and biologically relevant species and has been successfully applied for imaging of VDPs in living cells. We believe that the present pK a shift switching strategy may facilitate the development of new fluorescent probes that are useful for a wide range of applications.

  19. Fluorescence resonance energy transfer between conjugated molecules infiltrated in three-dimensional opal photonic crystals

    Zou, Lu; Sui, Ning; Wang, Ying-Hui; Qian, Cheng; Ma, Yu-Guang; Zhang, Han-Zhuang

    2015-01-01

    Fluorescence resonance energy transfer (FRET) from Coumarin 6 (C-6) to Sulforhodamine B (S-B) infiltrated into opal PMMA (poly-methyl-methacrylate) photonic crystals (PCs) has been studied in detail. The intrinsic mesh micro-porous structure of opal PCs could increase the luminescent efficiency through inhibiting the intermolecular interaction. Meanwhile, its structure of periodically varying refractive indices could also modify the FRET through affecting the luminescence characteristics of energy donor or energy acceptor. The results demonstrate that the FRET efficiency between conjugated dyes was easily modified by opal PCs. - Highlights: • We investigate the fluorescence resonance energy transfer between two kinds of dyes. • These two kinds of dyes are infiltrated in PMMA opal photonic crystals. • The structure of opal PCs could improve the luminescent characteristics. • The structure of opal PCs could improve the energy transfer characteristics

  20. Resonance fluorescence spectrum in a two-band photonic bandgap crystal

    Lee, Ray-Kuang; Lai, Yinchieh

    2003-05-01

    Steady state resonance fluorescence spectra from a two-level atom embedded in a photonic bandgap crystal and resonantly driven by a classical pump light are calculated. The photonic crystal is considered to be with a small bandgap which is in the order of magnitude of the Rabi frequency and is modeled by the anisotropic two-band dispersion relation. Non-Markovian noises caused by the non-uniform distribution of photon density states near the photonic bandgap are taken into account by a new approach which linearizes the optical Bloch equations by using the Liouville operator expansion. Fluorescence spectra that only exhibit sidebands of the Mollow triplet are found, indicating that there is no coherent Rayleigh scattering process.

  1. Fluorescence resonance energy transfer sensors for quantitative monitoring of pentose and disaccharide accumulation in bacteria

    Looger Loren L

    2008-06-01

    Full Text Available Abstract Background Engineering microorganisms to improve metabolite flux requires detailed knowledge of the concentrations and flux rates of metabolites and metabolic intermediates in vivo. Fluorescence resonance energy transfer sensors represent a promising technology for measuring metabolite levels and corresponding rate changes in live cells. These sensors have been applied successfully in mammalian and plant cells but potentially could also be used to monitor steady-state levels of metabolites in microorganisms using fluorimetric assays. Sensors for hexose and pentose carbohydrates could help in the development of fermentative microorganisms, for example, for biofuels applications. Arabinose is one of the carbohydrates to be monitored during biofuels production from lignocellulose, while maltose is an important degradation product of starch that is relevant for starch-derived biofuels production. Results An Escherichia coli expression vector compatible with phage λ recombination technology was constructed to facilitate sensor construction and was used to generate a novel fluorescence resonance energy transfer sensor for arabinose. In parallel, a strategy for improving the sensor signal was applied to construct an improved maltose sensor. Both sensors were expressed in the cytosol of E. coli and sugar accumulation was monitored using a simple fluorimetric assay of E. coli cultures in microtiter plates. In the case of both nanosensors, the addition of the respective ligand led to concentration-dependent fluorescence resonance energy transfer responses allowing quantitative analysis of the intracellular sugar levels at given extracellular supply levels as well as accumulation rates. Conclusion The nanosensor destination vector combined with the optimization strategy for sensor responses should help to accelerate the development of metabolite sensors. The new carbohydrate fluorescence resonance energy transfer sensors can be used for in vivo

  2. Simulation of fluorescence resonance energy transfer experiments: effect of the dyes on protein folding

    Allen, Lucy R; Paci, Emanuele

    2010-01-01

    Fluorescence resonance energy transfer is a powerful technique which is often used to probe the properties of proteins and complex macromolecules. The technique relies on relatively large fluorescent dyes which are engineered into the molecule of interest. In the case of small proteins, these dyes may affect the stability of the protein, and modify the folding kinetics and the folding mechanisms which are being probed. Here we use atomistic simulation to investigate the effect that commonly used fluorescent dyes have on the folding of a four-helix bundle protein. We show that, depending on where the dyes are attached, their effect on the kinetic and thermodynamic properties of the protein may be significant. We find that, while the overall folding mechanism is not affected by the dyes, they can destabilize, or even stabilize, intermediate states.

  3. Development of a dielectrophoresis-assisted surface plasmon resonance fluorescence biosensor for detection of bacteria

    Kuroda, Chiaki; Iizuka, Ryota; Ohki, Yoshimichi; Fujimaki, Makoto

    2018-05-01

    To detect biological substances such as bacteria speedily and accurately, a dielectrophoresis-assisted surface plasmon resonance (SPR) fluorescence biosensor is being developed. Using Escherichia coli as a target organism, an appropriate voltage frequency to collect E. coli cells on indium tin oxide quadrupole electrodes by dielectrophoresis is analyzed. Then, E. coli is stained with 4‧,6-diamidino-2-phenylindole (DAPI). To clearly detect fluorescence signals from DAPI-stained E. coli cells, the sensor is optimized so that we can excite SPR on Al electrodes by illuminating 405 nm photons. As a result, the number of fluorescence signals is increased on the electrodes by the application of a low-frequency voltage. This indicates that E. coli cells with a lower permittivity than the surrounding water are collected by negative dielectrophoresis onto the electrodes where the electric field strength is lowest.

  4. Intrinsic Tryptophan Fluorescence in the Detection and Analysis of Proteins: A Focus on Förster Resonance Energy Transfer Techniques

    Amar B. T. Ghisaidoobe

    2014-12-01

    Full Text Available F resonance energy transfer (FRET occurs when the distance between a donor fluorophore and an acceptor is within 10 nm, and its application often necessitates fluorescent labeling of biological targets. However, covalent modification of biomolecules can inadvertently give rise to conformational and/or functional changes. This review describes the application of intrinsic protein fluorescence, predominantly derived from tryptophan (\\(\\uplambda_{\\textsc{ex}}\\sim\\ nm, \\(\\uplambda_{\\textsc{em}}\\sim\\ 350 nm, in protein-related research and mainly focuses on label-free FRET techniques. In terms of wavelength and intensity, tryptophan fluorescence is strongly influenced by its (or the proteinlocal environment, which, in addition to fluorescence quenching, has been applied to study protein conformational changes. Intrinsic F resonance energy transfer (iFRET, a recently developed technique, utilizes the intrinsic fluorescence of tryptophan in conjunction with target-specific fluorescent probes as FRET donors and acceptors, respectively, for real time detection of native proteins.

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

    Meng-Chang Hsieh

    2015-02-01

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

  6. Fluorescence resonance energy transfer: A promising tool for investigation of the interaction between 1-anthracene sulphonate and serum albumins

    Banerjee, Paltu; Ghosh, Saptaparni; Sarkar, Arindam; Bhattacharya, Subhash Chandra

    2011-01-01

    This present investigation has revealed that steady state as well as time-resolved fluorescence techniques can serve as highly sensitive monitors for exploring the interaction of fluorescent probe 1-anthracene sulphonate (1-AS) with model transport proteins, bovine serum albumin (BSA) and human serum albumin (HSA).We have focused on fluorescence resonance energy transfer (FRET) between excited tryptophan in transport proteins to 1-AS, for the study of relaxation dynamics of biological molecules.

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

    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.

  8. Probing symmetry and symmetry breaking in resonant soft-x-ray fluorescence spectra of molecules

    Glans, P.; Gunnelin, K.; Guo, J. [Uppsala Univ. (Sweden)] [and others

    1997-04-01

    Conventional non-resonant soft X-ray emission brings about information about electronic structure through its symmetry and polarization selectivity, the character of which is governed by simple dipole rules. For centro-symmetric molecules with the emitting atom at the inversion center these rules lead to selective emission through the required parity change. For the more common classes of molecules which have lower symmetry or for systems with degenerate core orbitals (delocalized over identical sites), it is merely the local symmetry selectivity that provides a probe of the local atomic orbital contribution to the molecular orbital. For instance, in X-ray spectra of first row species the intensities essentially map the p-density at each particular atomic site, and, in a molecular orbital picture, the contribution of the local p-type atomic orbitals in the LCAO description of the molecular orbitals. The situation is different for resonant X-ray fluorescence spectra. Here strict parity and symmetry selectivity gives rise to a strong frequency dependence for all molecules with an element of symmetry. In addition to symmetry selectivity the strong frequency dependence of resonant X-ray emission is caused by the interplay between the shape of a narrow X-ray excitation energy function and the lifetime and vibrational broadenings of the resonantly excited core states. This interplay leads to various observable effects, such as linear dispersion, resonance narrowing and emission line (Stokes) doubling. Also from the point of view of polarization selectivity, the resonantly excited X-ray spectra are much more informative than the corresponding non-resonant spectra. Examples are presented for nitrogen, oxygen, and carbon dioxide molecules.

  9. Tunneling induced dark states and the controllable resonance fluorescence spectrum in quantum dot molecules

    Tian, Si-Cong; Tong, Cun-Zhu; Ning, Yong-Qiang; Qin, Li; Liu, Yun; Wan, Ren-Gang

    2014-01-01

    Optical spectroscopy, a powerful tool for probing and manipulating quantum dots (QDs), has been used to investigate the resonance fluorescence spectrum from linear triple quantum dot molecules controlled by tunneling, using atomic physics methods. Interesting features such as quenching and narrowing of the fluorescence are observed. In such molecules the tunneling between the quantum dots can also induce a dark state. The results are explained by the transition properties of the dressed states generated by the coupling of the laser and the tunneling. Unlike the atomic system, in such quantum dot molecules quantum coherence can be induced using tunneling, requiring no coupling lasers, which will allow tunneling controllable quantum dot molecules to be applied to quantum optics and photonics. (paper)

  10. Resonance fluorescence spectra of three-level atoms in a squeezed vacuum

    Ferguson, M.R.; Ficek, Z.; Dalton, B.J.

    1996-01-01

    The fluorescence field from one of the two allowed transitions in a three-level atom can sense squeezed fluctuations of a vacuum field coupled to the other transition. We examine the fluorescence spectra of strongly driven three-level atoms in Λ, V, and cascade configurations in which one of the two one-photon transitions is coupled to a finite-bandwidth squeezed vacuum field, when the bandwidth is much smaller than the difference in the atomic transition frequencies, though much larger than atomic decay rates and Rabi frequencies of the driving fields. The driving fields are on one-photon resonance, and the squeezed vacuum field is generated by a degenerate parameter oscillator. Details are only given for the Λ configuration. The extension to the V and cascade configurations is straightforward. We find that in all configurations the fluorescence spectra of the transition not coupled to the squeezed vacuum field are composed of five lines, one central and two pairs of sidebands, with intensities and widths strongly influenced by the squeezed vacuum field. However, only the central component and the outer sidebands exhibit a dependence on the squeezing phase. We also examine the fluorescence spectrum for the cascade configuration with a squeezed vacuum field on resonance with the two-photon transition between the ground and the most excited states and now generated by a nondegenerate parametric oscillator. In this case, where the squeezed vacuum field can be made coupled to both transitions, all spectral lines depend on the squeezing phase. The spectral features are explained in terms of the dressed-atom model of the system. We show that the coherent mixing of the atomic states by the strong driving fields modifies transition rates between the dressed states, which results in the selective phase dependence of the spectral features. copyright 1996 The American Physical Society

  11. Laser-excited Fluorescence And Electron-spin Resonance Of Er3+ In Polycrystalline Alcl3

    Ceotto G.; Pires M.A.; Sanjurjo J.A.; Rettori C.; Barberis G.E.

    1990-01-01

    The green fluorescence transitions among the levels corresponding to the 4S3/2 and 4I15/2 configurations of Er3+ diluted in AlCl3 have been measured using laser excitation. The data allow us to determine the crystalline-field splittings of these levels and, in turn, the spin-Hamiltonian parameters. The electron-paramagnetic-resonance spectrum observed at low temperatures is in good agreement with that expected from these parameters. © 1990 The American Physical Society.

  12. Measurement of changes in nuclear charge radii of 2r by laser-induced resonance fluorescence

    Gangrskij, Yu.P.; Zemlyanoj, S.G.; Marinova, K.P.; Markov, B.N.; Khoang Tkhi Kim Khueh; Chan Kong Tam; Kul'dzhanov, B.K.

    1987-01-01

    The optical isotopic shifts of Zr stable isotopes have been measured in three atomic transitions of type 4d 2 5s 2 → 4d 2 5s5p using the technique of laser-induced resonance fluorescence. The changes of nuclear mean-square charge radius Δ 2 > have been determined. The extracted values of Δ 2 > are compared to predictions of the droplet model. It is shown that the droplet model calculations can be made to agree with the experimental results, if changes of nuclear dynamical octupole deformation and of surface diffuseness parameter are taken into account

  13. Sub-Poissonian photon statistics in time-dependent collective resonance fluorescence

    Buzek, V.; Tran Quang; Lan, L.H.

    1989-10-01

    We have discussed the photon statistics of the spectral components of N-atom time-dependent resonance fluorescence. It is shown that in contrast to the stationary limit, sub-Poissonian photon statistics in the sidebands occur for any number N of atoms including the case N >> 1. Reduction in Maldel's parameters Q ±1 is found with increasing numbers of atoms. The typical time for the presence of sub-Poissonian statistics is proportional to 1/N. (author). 31 refs, 1 fig

  14. Resonance Fluorescence of a Trapped Four-Level Atom with Bichromatic Driving

    Bergou, J.; Jakob, M.; Abranyos, Y.

    1999-01-01

    The resonance fluorescence spectrum of a bichromatically driven four-level atom is polarization dependent. Very narrow lines occur in the incoherent parts of the spectrum for polarization directions which are different from that of the driving fields. The degree of squeezing has a maximum of 56% which should make it easily observable. The second-order correlation function exhibits anti bunching for zero time delay and strong super bunching for certain values of the interaction parameter and time delay. For these parameters resonant two-photon emission takes place in the form of polarization entangled photon pairs. The system can be a novel source of photons in the EPR and/or Bell states. Some experiments will be proposed which make use of this unique source. (Authors)

  15. Searching for illicit materials using nuclear resonance fluorescence stimulated by narrow-band photon sources

    Johnson, M.S., E-mail: johnson329@llnl.gov [Lawrence Livermore National Laboratory, Livermore, CA 94550 (United States); San Jose State University, San Jose, CA 95192 (United States); Hagmann, C.A.; Hall, J.M.; McNabb, D.P. [Lawrence Livermore National Laboratory, Livermore, CA 94550 (United States); Kelley, J.H.; Huibregtse, C. [North Carolina State University, Raleigh, NC 27695 (United States); Kwan, E.; Rusev, G.; Tonchev, A.P. [Duke University, Durham, NC 27708 (United States)

    2012-08-15

    We report the results of an experimental study of the sensitivity of two distinct classes of systems that exploit nuclear resonance fluorescence (NRF) to search for illicit materials in containers. One class of systems is based on the direct detection of NRF photons emitted from isotopes of interest. The other class infers the presence of a particular isotope by observing the preferential attenuation of resonant photons in the incident beam. We developed a detailed analytical model for both approaches. We performed experiments to test the model using depleted uranium as a surrogate for illicit material and used tungsten as a random choice for shielding. We performed the experiments at Duke University's High Intensity Gamma Source (HIGS). Using the methodology we detail in this paper one can use this model to estimate the performance of potential inspection systems in certifying containers as free of illicit materials and for detecting the presence of those same materials.

  16. Investigation of Membrane Receptors' Oligomers Using Fluorescence Resonance Energy Transfer and Multiphoton Microscopy in Living Cells

    Mishra, Ashish K.

    Investigating quaternary structure (oligomerization) of macromolecules (such as proteins and nucleic acids) in living systems (in vivo) has been a great challenge in biophysics, due to molecular diffusion, fluctuations in several biochemical parameters such as pH, quenching of fluorescence by oxygen (when fluorescence methods are used), etc. We studied oligomerization of membrane receptors in living cells by means of Fluorescence (Forster) Resonance Energy Transfer (FRET) using fluorescent markers and two photon excitation fluorescence micro-spectroscopy. Using suitable FRET models, we determined the stoichiometry and quaternary structure of various macromolecular complexes. The proteins of interest for this work are : (1) sigma-1 receptor and (2) rhodopsin, are described as below. (1) Sigma-1 receptors are molecular chaperone proteins, which also regulate ion channels. S1R seems to be involved in substance abuse, as well as several diseases such as Alzheimer's. We studied S1R in the presence and absence of its ligands haloperidol (an antagonist) and pentazocine +/- (an agonist), and found that at low concentration they reside as a mixture of monomers and dimers and that they may form higher order oligomers at higher concentrations. (2) Rhodopsin is a prototypical G protein coupled receptor (GPCR) and is directly involved in vision. GPCRs form a large family of receptors that participate in cell signaling by responding to external stimuli such as drugs, thus being a major drug target (more than 40% drugs target GPCRs). Their oligomerization has been largely controversial. Understanding this may help to understand the functional role of GPCRs oligomerization, and may lead to the discovery of more drugs targeting GPCR oligomers. It may also contribute toward finding a cure for Retinitis Pigmentosa, which is caused by a mutation (G188R) in rhodopsin, a disease which causes blindness and has no cure so far. Comparing healthy rhodopsin's oligomeric structure with that

  17. Isotopic imaging via nuclear resonance fluorescence with laser-based Thomson radiation

    Barty, Christopher P. J. [Hayward, CA; Hartemann, Frederic V [San Ramon, CA; McNabb, Dennis P [Alameda, CA; Pruet, Jason A [Brentwood, CA

    2009-07-21

    The present invention utilizes novel laser-based, high-brightness, high-spatial-resolution, pencil-beam sources of spectrally pure hard x-ray and gamma-ray radiation to induce resonant scattering in specific nuclei, i.e., nuclear resonance fluorescence. By monitoring such fluorescence as a function of beam position, it is possible to image in either two dimensions or three dimensions, the position and concentration of individual isotopes in a specific material configuration. Such methods of the present invention material identification, spatial resolution of material location and ability to locate and identify materials shielded by other materials, such as, for example, behind a lead wall. The foundation of the present invention is the generation of quasimonochromatic high-energy x-ray (100's of keV) and gamma-ray (greater than about 1 MeV) radiation via the collision of intense laser pulses from relativistic electrons. Such a process as utilized herein, i.e., Thomson scattering or inverse-Compton scattering, produces beams having diameters from about 1 micron to about 100 microns of high-energy photons with a bandwidth of .DELTA.E/E of approximately 10E.sup.-3.

  18. Resonant Scanning with Large Field of View Reduces Photobleaching and Enhances Fluorescence Yield in STED Microscopy.

    Wu, Yong; Wu, Xundong; Lu, Rong; Zhang, Jin; Toro, Ligia; Stefani, Enrico

    2015-10-01

    Photobleaching is a major limitation of superresolution Stimulated Depletion Emission (STED) microscopy. Fast scanning has long been considered an effective means to reduce photobleaching in fluorescence microscopy, but a careful quantitative study of this issue is missing. In this paper, we show that the photobleaching rate in STED microscopy can be slowed down and the fluorescence yield be enhanced by scanning with high speed, enabled by using large field of view in a custom-built resonant-scanning STED microscope. The effect of scanning speed on photobleaching and fluorescence yield is more remarkable at higher levels of depletion laser irradiance, and virtually disappears in conventional confocal microscopy. With ≥6 GW∙cm(-2) depletion irradiance, we were able to extend the fluorophore survival time of Atto 647N and Abberior STAR 635P by ~80% with 8-fold wider field of view. We confirm that STED Photobleaching is primarily caused by the depletion light acting upon the excited fluorophores. Experimental data agree with a theoretical model. Our results encourage further increasing the linear scanning speed for photobleaching reduction in STED microscopy.

  19. Distance distributions of short polypeptides recovered by fluorescence resonance energy transfer in the 10 A domain.

    Sahoo, Harekrushna; Roccatano, Danilo; Zacharias, Martin; Nau, Werner M

    2006-06-28

    Fluorescence resonance energy transfer (FRET) between tryptophan (Trp) as donor and 2,3-diazabicyclo[2.2.2]oct-2-ene (Dbo) as acceptor was studied by steady-state and time-resolved fluorescence spectroscopy. The unique feature of this FRET pair is its exceptionally short Förster radius (10 A), which allows one to recover distance distributions in very short structureless peptides. The technique was applied to Trp-(GlySer)n-Dbo-NH2 peptides with n = 0-10, for which the average probe/quencher distance ranged between 8.7 and 13.7 A experimentally (in propylene glycol, analysis according to wormlike chain model) and 8.6-10.2 A theoretically (for n = 0-6, GROMOS96 molecular dynamics simulations). The larger FRET efficiency in steady-state compared to time-resolved fluorescence experiments was attributed to a static quenching component, suggesting that a small but significant part (ca. 10%) of the conformations are already in van der Waals contact when excitation occurs.

  20. Probing the graphite band structure with resonant soft-x-ray fluorescence

    Carlisle, J.A.; Shirley, E.L.; Hudson, E.A. [Lawrence Berkeley National Lab., CA (United States)] [and others

    1997-04-01

    Soft x-ray fluorescence (SXF) spectroscopy using synchrotron radiation offers several advantages over surface sensitive spectroscopies for probing the electronic structure of complex multi-elemental materials. Due to the long mean free path of photons in solids ({approximately}1000 {angstrom}), SXF is a bulk-sensitive probe. Also, since core levels are involved in absorption and emission, SXF is both element- and angular-momentum-selective. SXF measures the local partial density of states (DOS) projected onto each constituent element of the material. The chief limitation of SXF has been the low fluorescence yield for photon emission, particularly for light elements. However, third generation light sources, such as the Advanced Light Source (ALS), offer the high brightness that makes high-resolution SXF experiments practical. In the following the authors utilize this high brightness to demonstrate the capability of SXF to probe the band structure of a polycrystalline sample. In SXF, a valence emission spectrum results from transitions from valence band states to the core hole produced by the incident photons. In the non-resonant energy regime, the excitation energy is far above the core binding energy, and the absorption and emission events are uncoupled. The fluorescence spectrum resembles emission spectra acquired using energetic electrons, and is insensitive to the incident photon`s energy. In the resonant excitation energy regime, core electrons are excited by photons to unoccupied states just above the Fermi level (EF). The absorption and emission events are coupled, and this coupling manifests itself in several ways, depending in part on the localization of the empty electronic states in the material. Here the authors report spectral measurements from highly oriented pyrolytic graphite.

  1. Parallel ion flow velocity measurement using laser induced fluorescence method in an electron cyclotron resonance plasma

    Yoshimura, Shinji; Okamoto, Atsushi; Terasaka, Kenichiro; Ogiwara, Kohei; Tanaka, Masayoshi Y.; Aramaki, Mitsutoshi

    2010-01-01

    Parallel ion flow velocity along a magnetic field has been measured using a laser induced fluorescence (LIF) method in an electron cyclotron resonance (ECR) argon plasma with a weakly-diverging magnetic field. To measure parallel flow velocity in a cylindrical plasma using the LIF method, the laser beam should be injected along device axis; however, the reflection of the incident beam causes interference between the LIF emission of the incident and reflected beams. Here we present a method of quasi-parallel laser injection at a small angle, which utilizes the reflected beam as well as the incident beam to obtain the parallel ion flow velocity. Using this method, we observed an increase in parallel ion flow velocity along the magnetic field. The acceleration mechanism is briefly discussed on the basis of the ion fluid model. (author)

  2. Structure and dynamics of olefin radical cation aggregates. Time-resolved fluorescence detected magnetic resonance

    Desrosiers, M.F.; Trifunac, A.D.

    1986-01-01

    The time-resolved EPR spectra and thus the structure and dynamics of transient hydrocarbon radical cations are obtained by the pulse radiolysis-fluorescence detected magnetic resonance (FDMR) technique. Here the authors report the observation of short-lived radical cations from olefins. FDMR-EPR spectra of radical cations from tetramethylethylene and cyclohexadiene are illustrated. The olefin radical cations, FDMR spectra are concentration-dependent, since dimerization with neutral molecules takes place at higher (>10 -2 M) olefin concentration. Rate constants for the dimerization reaction are derived and the effect of solvent viscosity on aggregate formation is demonstrated. By monitoring the further reactions of dimer cations the authors have obtained EPR evidence for previously unobserved higher-order (multimer) radical cation aggregates of olefins. 16 references, 5 figures

  3. The use of Fluorescence Resonance Energy Transfer (FRET peptidesfor measurement of clinically important proteolytic enzymes

    Adriana K. Carmona

    2009-09-01

    Full Text Available Proteolytic enzymes have a fundamental role in many biological processes and are associated with multiple pathological conditions. Therefore, targeting these enzymes may be important for a better understanding of their function and development of therapeutic inhibitors. Fluorescence Resonance Energy Transfer (FRET peptides are convenient tools for the study of peptidases specificity as they allow monitoring of the reaction on a continuous basis, providing a rapid method for the determination of enzymatic activity. Hydrolysis of a peptide bond between the donor/acceptor pair generates fluorescence that permits the measurement of the activity of nanomolar concentrations of the enzyme. The assays can be performed directly in a cuvette of the fluorimeter or adapted for determinations in a 96-well fluorescence plate reader. The synthesis of FRET peptides containing ortho-aminobenzoic acid (Abz as fluorescent group and 2, 4-dinitrophenyl (Dnp or N-(2, 4-dinitrophenylethylenediamine (EDDnp as quencher was optimized by our group and became an important line of research at the Department of Biophysics of the Federal University of São Paulo. Recently, Abz/Dnp FRET peptide libraries were developed allowing high-throughput screening of peptidases substrate specificity. This review presents the consolidation of our research activities undertaken between 1993 and 2008 on the synthesis of peptides and study of peptidases specificities.As enzimas proteolíticas têm um papel fundamental em muitos processos biológicos e estão associadas a vários estados patológicos. Por isso, o estudo da especificidade das peptidases pode ser importante para uma melhor compreensão da função destas enzimas e para o desenvolvimento de inibidores. Os substratos com supressão intramolecular de fluorescência constituem uma excelente ferramenta, pois permitem o monitoramento da reação de forma contínua, proporcionando um método prático e rápido para a determinação da

  4. Resonance energy transfer based electrochemiluminescence and fluorescence sensing of riboflavin using graphitic carbon nitride quantum dots

    Wang, Huan [Key Laboratory of Bioelectrochemistry & Environmental Analysis of Gansu Province, College of Chemistry & Chemical Engineering, Northwest Normal University, Lanzhou, Gansu 730070 (China); The Phytochemistry Key Laboratory of Tibetan Plateau of Qinghai Province, College of Pharmacy, Qinghai Nationalities University, Xining, Qinghai 810007 (China); Ma, Qin; Wang, Yanfeng; Wang, Caihe; Qin, Dongdong; Shan, Duoliang; Chen, Jing [Key Laboratory of Bioelectrochemistry & Environmental Analysis of Gansu Province, College of Chemistry & Chemical Engineering, Northwest Normal University, Lanzhou, Gansu 730070 (China); Lu, Xiaoquan, E-mail: luxq@nwnu.edu.cn [Key Laboratory of Bioelectrochemistry & Environmental Analysis of Gansu Province, College of Chemistry & Chemical Engineering, Northwest Normal University, Lanzhou, Gansu 730070 (China)

    2017-06-22

    Graphitic carbon nitride quantum dots (g-CNQDs) are rarely used in the field of electrochemiluminescence. In this paper, g-CNQDs have a strong and stable electrochemiluminescence (ECL) signal generated in the presence of co-reactant K{sub 2}S{sub 2}O{sub 8}. The ECL signal of g-CNQDs was quenched by the mechanism of resonance energy transfer (RET) between donor g-CNQDs and receptor riboflavin (RF) that is proved by UV-vis absorption spectroscopy, electrochemiluminescence and fluorescence emission spectroscopy analysis technology. Therefore, we achieved detection of the riboflavin content in the drug tablets of vitamin B{sub 2} using ECL and FL. The determination results of ECL showed that the riboflavin content of the drug vitamin B{sub 2} (VB{sub 2}) tablets was consistent with the fluorescence (FL) analysis, with wider linear range of 0.02–11 μM and lower minimum detection limit of 0.63 nM (S/N = 3) than FL. Hence, the riboflavin content in human serum was further detected using ECL. The relative standard deviation is less than 6.5%, with an acceptable recovery of 95.33%–104.22%, which means that this sensor has potential applications in the actual sample analysis. As a new ECL luminary, g-CNQDs have opened a new field for the development and application of ECL sensor. - Highlights: • G-CNQDs proposed as a new luminophore for ECL. • ECL signal was strong and stable in the presence of co-reactant K{sub 2}S{sub 2}O{sub 8}. • Based on the resonance energy transfer between g-CNQDs and riboflavin. • ECL has wider linear range and lower detection limit than FL.

  5. Resonance energy transfer based electrochemiluminescence and fluorescence sensing of riboflavin using graphitic carbon nitride quantum dots

    Wang, Huan; Ma, Qin; Wang, Yanfeng; Wang, Caihe; Qin, Dongdong; Shan, Duoliang; Chen, Jing; Lu, Xiaoquan

    2017-01-01

    Graphitic carbon nitride quantum dots (g-CNQDs) are rarely used in the field of electrochemiluminescence. In this paper, g-CNQDs have a strong and stable electrochemiluminescence (ECL) signal generated in the presence of co-reactant K 2 S 2 O 8 . The ECL signal of g-CNQDs was quenched by the mechanism of resonance energy transfer (RET) between donor g-CNQDs and receptor riboflavin (RF) that is proved by UV-vis absorption spectroscopy, electrochemiluminescence and fluorescence emission spectroscopy analysis technology. Therefore, we achieved detection of the riboflavin content in the drug tablets of vitamin B 2 using ECL and FL. The determination results of ECL showed that the riboflavin content of the drug vitamin B 2 (VB 2 ) tablets was consistent with the fluorescence (FL) analysis, with wider linear range of 0.02–11 μM and lower minimum detection limit of 0.63 nM (S/N = 3) than FL. Hence, the riboflavin content in human serum was further detected using ECL. The relative standard deviation is less than 6.5%, with an acceptable recovery of 95.33%–104.22%, which means that this sensor has potential applications in the actual sample analysis. As a new ECL luminary, g-CNQDs have opened a new field for the development and application of ECL sensor. - Highlights: • G-CNQDs proposed as a new luminophore for ECL. • ECL signal was strong and stable in the presence of co-reactant K 2 S 2 O 8 . • Based on the resonance energy transfer between g-CNQDs and riboflavin. • ECL has wider linear range and lower detection limit than FL.

  6. Study on the fluorescence resonance energy transfer between CdS quantum dots and Eosin Y.

    Yan, Zhengyu; Zhang, Zhengwei; Yu, Yan; Chen, Jianqiu

    2015-03-01

    Water-soluble CdS quantum dots (QDs) were prepared using mercaptoacetic acid (TGA) as the stabilizer in an aqueous system. A fluorescence resonance energy transfer (FRET) system was constructed between water-soluble CdS QDs (donor) and Eosin Y (acceptor). Several factors that impacted the fluorescence spectra of the FRET system, such as pH (3.05-10.10), concentration of Eosin Y (2-80 mg/L) and concentration of CdS QDs (2-80 mg/L), were investigated and refined. Donor-to-acceptor ratios, the energy transfer efficiency (E) and the distance (r) between CdS QDs and Eosin Y were obtained. The results showed that a FRET system could be established between water-soluble CdS QDs and Eosin Y at pH 5.0; donor-to-acceptor ratios demonstrated a 1: 8 proportion of complexes; the energy transfer efficiency (E) and the distance (r) between the QDs and Eosin Y were 20.07% and 4.36 nm,respectively. Copyright © 2014 John Wiley & Sons, Ltd.

  7. Development of L-lactate dehydrogenase biosensor based on porous silicon resonant microcavities as fluorescence enhancers.

    Jenie, S N Aisyiyah; Prieto-Simon, Beatriz; Voelcker, Nicolas H

    2015-12-15

    The up-regulation of L-lactate dehydrogenase (LDH), an intracellular enzyme present in most of all body tissues, is indicative of several pathological conditions and cellular death. Herein, we demonstrate LDH detection using porous silicon (pSi) microcavities as a luminescence-enhancing optical biosensing platform. Non-fluorescent resazurin was covalently attached onto the pSi surface via thermal hydrocarbonisation, thermal hydrosylilation and acylation. Each surface modification step was confirmed by means of FTIR and the optical shifts of the resonance wavelength of the microcavity. Thermal hydrocarbonisation also afforded excellent surface stability, ensuring that the resazurin was not reduced on the pSi surface. Using a pSi microcavity biosensor, the fluorescence signal upon detection of LDH was amplified by 10 and 5-fold compared to that of a single layer and a detuned microcavity, respectively, giving a limit of detection of 0.08 U/ml. The biosensor showed a linear response between 0.16 and 6.5 U/ml, covering the concentration range of LDH in normal as well as damaged tissues. The biosensor was selective for LDH and did not produce a signal upon incubation with another NAD-dependant enzyme L-glutamic dehydrogenase. The use of the pSi microcavity as a sensing platform reduced reagent usage by 30% and analysis time threefold compared to the standard LDH assay in solution. Copyright © 2015 Elsevier B.V. All rights reserved.

  8. Fluorescence resonance energy transfer (FRET-based subcellular visualization of pathogen-induced host receptor signaling

    Zimmermann Timo

    2009-11-01

    Full Text Available Abstract Background Bacteria-triggered signaling events in infected host cells are key elements in shaping the host response to pathogens. Within the eukaryotic cell, signaling complexes are spatially organized. However, the investigation of protein-protein interactions triggered by bacterial infection in the cellular context is technically challenging. Here, we provide a methodological approach to exploit fluorescence resonance energy transfer (FRET to visualize pathogen-initiated signaling events in human cells. Results Live-cell microscopy revealed the transient recruitment of the Src family tyrosine kinase Hck upon bacterial engagement of the receptor carcinoembryonic antigen-related cell adhesion molecule 3 (CEACAM3. In cells expressing a CEACAM3 variant lacking the cytoplasmic domain, the Src homology 2 (SH2 domain of Hck (Hck-SH2 was not recruited, even though bacteria still bound to the receptor. FRET measurements on the basis of whole cell lysates revealed intimate binding between Hck-SH2 (using enhanced yellow fluorescent protein (YPet-Hck-SH2 and the tyrosine-phosphorylated enhanced cyan fluorescent protein-labeled cytoplasmic domain of wild-type CEACAM3 (CEACAM3 WT-CyPet and a flow cytometry-based FRET approach verified this association in intact cells. Using confocal microscopy and acceptor photobleaching, FRET between Hck-SH2 and CEACAM3 was localized to the sites of bacteria-host cell contact. Conclusion These data demonstrate not only the intimate binding of the SH2 domain of Hck to the tyrosine-phosphorylated cytoplasmic domain of CEACAM3 in intact cells, but furthermore, FRET measurements allow the subcellular localization of this process during bacterial infection. FRET-based assays are valuable tools to resolve bacteria-induced protein-protein interactions in the context of the intact host cell.

  9. Folic acid-targeted magnetic Tb-doped CeF3 fluorescent nanoparticles as bimodal probes for cellular fluorescence and magnetic resonance imaging.

    Ma, Zhi-Ya; Liu, Yu-Ping; Bai, Ling-Yu; An, Jie; Zhang, Lin; Xuan, Yang; Zhang, Xiao-Shuai; Zhao, Yuan-Di

    2015-10-07

    Magnetic fluorescent nanoparticles (NPs) have great potential applications for diagnostics, imaging and therapy. We developed a facile polyol method to synthesize multifunctional Fe3O4@CeF3:Tb@CeF3 NPs with small size (CA) to obtain carboxyl-functionalized NPs (Fe3O4@CeF3:Tb@CeF3-COOH). Folic acid (FA) as an affinity ligand was then covalently conjugated onto NPs to yield Fe3O4@CeF3:Tb@CeF3-FA NPs. They were then applied as multimodal imaging agents for simultaneous in vitro targeted fluorescence imaging and magnetic resonance imaging (MRI) of HeLa cells with overexpressed folate receptors (FR). The results indicated that these NPs had strong luminescence and enhanced T2-weighted MR contrast and would be promising candidates as multimodal probes for both fluorescence and MRI imaging.

  10. Comparative in vivo mucoadhesion studies of thiomer formulations using magnetic resonance imaging and fluorescence detection.

    Albrecht, K; Greindl, M; Kremser, C; Wolf, C; Debbage, P; Bernkop-Schnürch, A

    2006-09-28

    The aim of this study was to compare different oral delivery systems based on the thiolated polymer polycarbophil-cysteine (PCP-Cys) and to provide evidence for the validity of the hypothesis that unhydrated polymers provide better mucoadhesion in vivo. To achieve dry polymer application, a new, experimental dosage form named Eutex (made of Eudragit L100-55 and latex) capsule has been developed. Magnetic resonance imaging was used to localize the point of release of the thiolated polymer from the application forms via the positive magnetic resonance signal from a gadolinium complex (Gd-DTPA). In vivo mucoadhesion was determined by ascertaining the residence time of the fluorescence-tagged thiomer on intestinal mucosa after 3 h. Results showed that in comparison to conventional application forms the Eutex capsules led to 1.9-fold higher mucoadhesive properties of PCP-Cys when compared to application with a conventional enteric-coated capsule, and to 1.4-fold higher mucoadhesion when compared to administration with an enteric-coated tablet of the thiomer. The findings of this study should contribute to the understanding of mucoadhesion and mucoadhesion influencing parameters in vivo and should therefore be of considerable interest for the development of future mucoadhesive oral drug delivery dosage forms.

  11. Resonant inelastic scattering in dilute magnetic semiconductors by x-ray fluorescence spectroscopy

    Lawniczak-Jablonska, K. [Lawrence Berkeley National Lab., CA (United States)]|[Institute of Physics, Warsaw (Poland); Jia, J.J.; Underwood, J.H. [Lawrence Berkeley National Lab., CA (United States)] [and others

    1997-04-01

    As modern, technologically important materials have become more complex, element specific techniques have become invaluable in studying the electronic structure of individual components from the system. Soft x-ray fluorescence (SXF) and absorption (SXA) spectroscopies provide a unique means of measuring element and angular momentum density of electron states, respectively, for the valence and conducting bands in complex materials. X-ray absorption and the decay through x-ray emission are generally assumed to be two independent one-photon processes. Recent studies, however have demonstrated that SXF excited near the absorption threshold generate an array of spectral features that depend on nature of materials, particularly on the localization of excited states in s and d-band solids and that these two processes can no be longer treated as independent. Resonant SXF offers thus the new way to study the dynamics of the distribution of electronic valence states in the presence of a hole which is bound to the electron low lying in the conduction band. This process can simulate the interaction between hole-electron pair in wide gap semiconductors. Therefore such studies can help in understanding of transport and optics phenomena in the wide gap semiconductors. The authors report the result of Mn and S L-resonant emission in Zn{sub 1{minus}x}Mn{sub x}S (with x=0.2 and 0.3) and MnS as the energy of exciting radiation is tuned across the Mn and S L{sub 3,2} absorption edge, along with the resonant excited spectra from elemental Mn as a reference.

  12. A reduced graphene oxide-based fluorescence resonance energy transfer sensor for highly sensitive detection of matrix metalloproteinase 2.

    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.

  13. Nuclear Resonance Fluorescence to Measure Plutonium Mass in Spent Nuclear Fuel

    Ludewigt, Bernhard A; Quiter, Brian J.; Ambers, Scott D.

    2011-01-14

    The Next Generation Safeguard Initiative (NGSI) of the U.S Department of Energy is supporting a multi-lab/university collaboration to quantify the plutonium (Pu) mass in spent nuclear fuel (SNF) assemblies and to detect the diversion of pins with non-destructive assay (NDA) methods. The following 14 NDA techniques are being studied: Delayed Neutrons, Differential Die-Away, Differential Die-Away Self-Interrogation, Lead Slowing Down Spectrometer, Neutron Multiplicity, Passive Neutron Albedo Reactivity, Total Neutron (Gross Neutron), X-Ray Fluorescence, {sup 252}Cf Interrogation with Prompt Neutron Detection, Delayed Gamma, Nuclear Resonance Fluorescence, Passive Prompt Gamma, Self-integration Neutron Resonance Densitometry, and Neutron Resonance Transmission Analysis. Understanding and maturity of the techniques vary greatly, ranging from decades old, well-understood methods to new approaches. Nuclear Resonance Fluorescence (NRF) is a technique that had not previously been studied for SNF assay or similar applications. Since NRF generates isotope-specific signals, the promise and appeal of the technique lies in its potential to directly measure the amount of a specific isotope in an SNF assay target. The objectives of this study were to design and model suitable NRF measurement methods, to quantify capabilities and corresponding instrumentation requirements, and to evaluate prospects and the potential of NRF for SNF assay. The main challenge of the technique is to achieve the sensitivity and precision, i.e., to accumulate sufficient counting statistics, required for quantifying the mass of Pu isotopes in SNF assemblies. Systematic errors, considered a lesser problem for a direct measurement and only briefly discussed in this report, need to be evaluated for specific instrument designs in the future. Also, since the technical capability of using NRF to measure Pu in SNF has not been established, this report does not directly address issues such as cost, size

  14. Statistical uncertainties of nondestructive assay for spent nuclear fuel by using nuclear resonance fluorescence

    Shizuma, Toshiyuki, E-mail: shizuma.toshiyuki@jaea.go.jp [Quantum Beam Science Directorate, Japan Atomic Energy Agency, Tokai, Ibaraki 319-1195 (Japan); Hayakawa, Takehito; Angell, Christopher T.; Hajima, Ryoichi [Quantum Beam Science Directorate, Japan Atomic Energy Agency, Tokai, Ibaraki 319-1195 (Japan); Minato, Futoshi; Suyama, Kenya [Nuclear Science and Engineering Directorate, Japan Atomic Energy Agency, Tokai, Ibaraki 319-1195 (Japan); Seya, Michio [Integrated Support Center for Nuclear Nonproliferation and Nuclear Security, Japan Atomic Energy Agency, Tokai, Ibaraki 319-1198 (Japan); Johnson, Micah S. [Lawrence Livermore National Laboratory, 7000 East Ave. Livermore, CA 94550 (United States); Department of Physics and Astronomy, San Jose State University, One Washington Square, San Jose, CA 9519 (United States); McNabb, Dennis P. [Lawrence Livermore National Laboratory, 7000 East Ave. Livermore, CA 94550 (United States)

    2014-02-11

    We estimated statistical uncertainties of a nondestructive assay system using nuclear resonance fluorescence (NRF) for spent nuclear fuel including low-concentrations of actinide nuclei with an intense, mono-energetic photon beam. Background counts from radioactive materials inside the spent fuel were calculated with the ORIGEN2.2-UPJ burn-up computer code. Coherent scattering contribution associated with Rayleigh, nuclear Thomson, and Delbrück scattering was also considered. The energy of the coherent scattering overlaps with that of NRF transitions to the ground state. Here, we propose to measure NRF transitions to the first excited state to avoid the coherent scattering contribution. Assuming that the total NRF cross-sections are in the range of 3–100 eV b at excitation energies of 2.25, 3.5, and 5 MeV, statistical uncertainties of the NRF measurement were estimated. We concluded that it is possible to assay 1% actinide content in the spent fuel with 2.2–3.2% statistical precision during 4000 s measurement time for the total integrated cross-section of 30 eV b at excitation energies of 3.5–5 MeV by using a photon beam with an intensity of 10{sup 6} photons/s/eV. We also examined both the experimental and theoretical NRF cross-sections for actinide nuclei. The calculation based on the quasi-particle random phase approximation suggests the existence of strong magnetic dipole resonances at excitation energies ranging from 2 to 6 MeV with the scattering cross-sections of tens eV b around 5 MeV in {sup 238}U.

  15. Fluorescence resonance energy transfer between perylene and riboflavin in micellar solution and analytical application on determination of vitamin B2

    Bhattar, S.L.; Kolekar, G.B.; Patil, S.R.

    2008-01-01

    Fluorescence resonance energy transfer (FRET) between perylene and riboflavin is studied in micellar solution of sodium dodecyl sulfate. The fluorescence of perylene is quenched by riboflavin and quenching is in accordance with Stern-Volmer relation. The efficiency of energy transfer is found to depend on the concentration of riboflavin. The value of critical energy transfer distance (R 0 ) calculated by using Foster relation is 32.13 A, and as it is less than 50 A, it indicates efficient energy transfer in the present system. The analytical relation was established between extent of sensitization and concentration of riboflavin, which helped to estimate vitamin B 2 directly from pharmaceutical tablets

  16. Smart Drug Delivery System-Inspired Enzyme-Linked Immunosorbent Assay Based on Fluorescence Resonance Energy Transfer and Allochroic Effect Induced Dual-Modal Colorimetric and Fluorescent Detection.

    Miao, Luyang; Zhu, Chengzhou; Jiao, Lei; Li, He; Du, Dan; Lin, Yuehe; Wei, Qin

    2018-02-06

    Numerous analytical techniques have been undertaken for the detection of protein biomarkers because of their extensive and significant applications in clinical diagnosis, whereas there are few strategies to develop dual-readout immunosensors to achieve more accurate results. To the best of our knowledge, inspired by smart drug delivery system (DDS), a novel pH-responsive modified enzyme-linked immunosorbent assay (ELISA) was innovatively developed for the first time, realizing dual-modal colorimetric and fluorescent detection of cardiac troponin I (cTnI). Curcumin (CUR) was elaborately selected as a reporter molecule, which played the same role of drugs in DDS based on the following considerations: (1) CUR can be used as a kind of pH indicator by the inherited allochroic effect induced by basic pH value; (2) the fluorescence of CUR can be quenched by certain nanocarriers as the acceptor because of the occurrence of fluorescence resonance energy transfer (FRET), while recovered by the stimuli of basic pH value, which can produce "signal-on" fluorescence detection. Three-dimensional MoS 2 nanoflowers (3D-MoS 2 NFs) were employed in immobilizing CUR to constitute a nanoprobe for the determination of cTnI by virtue of good biocompatibility, high absorption capacity, and fluorescence quench efficiency toward CUR. The proposed DDS-inspired ELISA offered dual-modal colorimetric and fluorescent detection of cTnI, thereby meeting the reliable and precise analysis requirements. We believe that the developed dual-readout ELISA will create a new avenue and bring innovative inspirations for biological detections.

  17. Iodinated oil-loaded, fluorescent mesoporous silica-coated iron oxide nanoparticles for magnetic resonance imaging/computed tomography/fluorescence trimodal imaging

    Xue S

    2014-05-01

    Full Text Available Sihan Xue,1 Yao Wang,1 Mengxing Wang,2 Lu Zhang,1 Xiaoxia Du,2 Hongchen Gu,1 Chunfu Zhang1,31School of Biomedical Engineering and Med-X Research Institute, Shanghai Jiao Tong University, 2Shanghai Key Laboratory of Magnetic Resonance, Department of Physics, East China Normal University, 3State Key Laboratory of Oncogenes and Related Genes, Shanghai Cancer Institute, School of Biomedical Engineering, Shanghai Jiao Tong University, Shanghai, People’s Republic of ChinaAbstract: In this study, a novel magnetic resonance imaging (MRI/computed tomography (CT/fluorescence trifunctional probe was prepared by loading iodinated oil into fluorescent mesoporous silica-coated superparamagnetic iron oxide nanoparticles (i-fmSiO4@SPIONs. Fluorescent mesoporous silica-coated superparamagnetic iron oxide nanoparticles (fmSiO4@SPIONs were prepared by growing fluorescent dye-doped silica onto superparamagnetic iron oxide nanoparticles (SPIONs directed by a cetyltrimethylammonium bromide template. As prepared, fmSiO4@SPIONs had a uniform size, a large surface area, and a large pore volume, which demonstrated high efficiency for iodinated oil loading. Iodinated oil loading did not change the sizes of fmSiO4@SPIONs, but they reduced the MRI T2 relaxivity (r2 markedly. I-fmSiO4@SPIONs were stable in their physical condition and did not demonstrate cytotoxic effects under the conditions investigated. In vitro studies indicated that the contrast enhancement of MRI and CT, and the fluorescence signal intensity of i-fmSiO4@SPION aqueous suspensions and macrophages, were intensified with increased i-fmSiO4@SPION concentrations in suspension and cell culture media. Moreover, for the in vivo study, the accumulation of i-fmSiO4@SPIONs in the liver could also be detected by MRI, CT, and fluorescence imaging. Our study demonstrated that i-fmSiO4@SPIONs had great potential for MRI/C/fluorescence trimodal imaging.Keywords: multifunctional probe, SPIONs, mesoporous silica

  18. Design and fabrication of fluorescence resonance energy transfer-mediated fluorescent polymer nanoparticles for ratiometric sensing of lysosomal pH.

    Chen, Jian; Tang, Ying; Wang, Hong; Zhang, Peisheng; Li, Ya; Jiang, Jianhui

    2016-12-15

    The design of effective tools capable of sensing lysosome pH is highly desirable for better understanding its biological functions in cellular behaviors and various diseases. Herein, a lysosome-targetable ratiometric fluorescent polymer nanoparticle pH sensor (RFPNS) was synthesized via incorporation of miniemulsion polymerization and surface modification technique. In this system, the donor: 4-ethoxy-9-allyl-1,8-naphthalimide (EANI) and the acceptor: fluorescein isothiocyanate (FITC) were covalently linked to the polymer nanoparticle to construct pH-responsive fluorescence resonance energy transfer (FRET) system. The FITC moieties on the surface of RFPNS underwent structural and spectral transformation as the presence of pH changes, resulting in ratiometric fluorescent sensing of pH. The as-prepared RFPNS displayed favorable water dispersibility, good pH-induced spectral reversibility and so on. Following the living cell uptake, the as-prepared RFPNS with good cell-membrane permeability can mainly stain in the lysosomes; and it can facilitate visualization of the intracellular lysosomal pH changes. This nanosensor platform offers a novel method for future development of ratiometric fluorescent probes for targeting other analytes, like ions, metabolites,and other biomolecules in biosamples. Copyright © 2016 Elsevier Inc. All rights reserved.

  19. A high-throughput direct fluorescence resonance energy transfer-based assay for analyzing apoptotic proteases using flow cytometry and fluorescence lifetime measurements.

    Suzuki, Miho; Sakata, Ichiro; Sakai, Takafumi; Tomioka, Hiroaki; Nishigaki, Koichi; Tramier, Marc; Coppey-Moisan, Maïté

    2015-12-15

    Cytometry is a versatile and powerful method applicable to different fields, particularly pharmacology and biomedical studies. Based on the data obtained, cytometric studies are classified into high-throughput (HTP) or high-content screening (HCS) groups. However, assays combining the advantages of both are required to facilitate research. In this study, we developed a high-throughput system to profile cellular populations in terms of time- or dose-dependent responses to apoptotic stimulations because apoptotic inducers are potent anticancer drugs. We previously established assay systems involving protease to monitor live cells for apoptosis using tunable fluorescence resonance energy transfer (FRET)-based bioprobes. These assays can be used for microscopic analyses or fluorescence-activated cell sorting. In this study, we developed FRET-based bioprobes to detect the activity of the apoptotic markers caspase-3 and caspase-9 via changes in bioprobe fluorescence lifetimes using a flow cytometer for direct estimation of FRET efficiencies. Different patterns of changes in the fluorescence lifetimes of these markers during apoptosis were observed, indicating a relationship between discrete steps in the apoptosis process. The findings demonstrate the feasibility of evaluating collective cellular dynamics during apoptosis. Copyright © 2015 Elsevier Inc. All rights reserved.

  20. Decay time shortening of fluorescence from donor-acceptor pair proteins using ultrafast time-resolved fluorescence resonance energy transfer spectroscopy

    Baba, Motoyoshi; Suzuki, Masayuki; Ganeev, Rashid A.; Kuroda, Hiroto; Ozaki, Tsuneyuki; Hamakubo, Takao; Masuda, Kazuyuki; Hayashi, Masahiro; Sakihama, Toshiko; Kodama, Tatsuhiko; Kozasa, Tohru

    2007-01-01

    We improved an ultrafast time-resolved fluorescence resonance energy transfer (FRET) spectroscopy system and measured directly the decrease in the fluorescence decay time of the FRET signal, without any entanglement of components in the picosecond time scale from the donor-acceptor protein pairs (such as cameleon protein for calcium ion indicator, and ligand-activated GRIN-Go proteins pair). The drastic decrease in lifetime of the donor protein fluorescence under the FRET condition (e.g. a 47.8% decrease for a GRIN-Go protein pair) proves the deformation dynamics between donor and acceptor fluorescent proteins in an activated state of a mixed donor-acceptor protein pair. This study is the first clear evidence of physical contact of the GRIN-Go proteins pair using time-resolved FRET system. G protein-coupled receptors (GPCRs) are the most important protein family for the recognition of many chemical substances at the cell surface. They are the targets of many drugs. Simultaneously, we were able to observe the time-resolved spectra of luminous proteins at the initial stage under the FRET condition, within 10 ns from excitation. This new FRET system allows us to trace the dynamics of the interaction between proteins at the ligand-induced activated state, molecular structure change and combination or dissociation. It will be a key technology for the development of protein chip technology

  1. Effect of membrane microheterogeneity and domain size on fluorescence resonance energy transfer.

    Towles, Kevin B; Brown, Angela C; Wrenn, Steven P; Dan, Nily

    2007-07-15

    Studies of multicomponent membranes suggest lateral inhomogeneity in the form of membrane domains, but the size of small (nanoscale) domains in situ cannot be determined with current techniques. In this article, we present a model that enables extraction of membrane domain size from time-resolved fluorescence resonance energy transfer (FRET) data. We expand upon a classic approach to the infinite phase separation limit and formulate a model that accounts for the presence of disklike domains of finite dimensions within a two-dimensional infinite planar bilayer. The model was tested against off-lattice Monte Carlo calculations of a model membrane in the liquid-disordered (l(d)) and liquid-ordered (l(o)) coexistence regime. Simulated domain size was varied from 5 to 50 nm, and two fluorophores, preferentially partitioning into opposite phases, were randomly mixed to obtain the simulated time-resolved FRET data. The Monte Carlo data show clear differences in the efficiency of energy transfer as a function of domain size. The model fit of the data yielded good agreement for the domain size, especially in cases where the domain diameter is membrane domains using time-resolved FRET.

  2. Comparison of different Bremsstrahlung converters and collimators for Nuclear Resonance Fluorescence setup at IFUSP

    Lopez, P.N; Corrales, Y.; Manso Guevara, M.V; Martins, M.N.

    2007-01-01

    Nuclear Resonance Fluorescence (NRF) setup will install in the new electron accelerator, which is in final stage of installation at the Physics Institute of Sao Paulo University (IFUSP). The Bremsstrahlung facility and the setup for photon scattering should be designed such that the background radiation caused by scattering photons and the production of neutrons is minimized. In this order the Monte Carlo simulation studies show the best options for the different elements of the NRF setup, and how to link these elements to the particularities of the irradiation room. In the present stage the simulations has been included the studies of different Bremsstrahlung converters and collimators. Several materials (Ta, W, Au, Nb, Cu) for Bremsstrahlung converters were studied. Detailed analyses of intensity as well as the opening angles of Bremsstrahlung radiation were carried out, for different converter thickness. For the collimator two materials (Cu and Pb) were studied in the simulations. Several opening angles and thickness (40 - 100 cm) were studied. The Bremsstrahlung beam collimation for different energy bins, and the photon scattering from the collimator ,were used as quality parameters of the collimators. (Author)

  3. Resonance fluorescence spectrum of a p-doped quantum dot coupled to a metallic nanoparticle

    Carreño, F.; Antón, M. A.; Arrieta-Yáñez, Francisco

    2013-11-01

    The resonance fluorescence spectrum (RFS) of a hybrid system consisting of a p-doped semiconductor quantum dot (QD) coupled to a metallic nanoparticle (MNP) is analyzed. The quantum dot is described as a four-level atomlike system using the density matrix formalism. The lower levels are Zeeman-split hole spin states and the upper levels correspond to positively charged excitons containing a spin-up, spin-down hole pair and a spin electron. A linearly polarized laser field drives two of the optical transitions of the QD and produces localized surface plasmons in the nanoparticle, which act back upon the QD. The frequencies of these localized plasmons are very different along the two principal axes of the nanoparticle, thus producing an anisotropic modification of the spontaneous emission rates of the allowed optical transitions, which is accompanied by very minor local field corrections. This manifests into dramatic modifications in the RFS of the hybrid system in contrast to the one obtained for the isolated QD. The RFS is analyzed as a function of the nanoparticle's aspect ratio, the external magnetic field applied in the Voigt geometry, and the Rabi frequency of the driving field. It is shown that the spin of the QD is imprinted onto certain sidebands of the RFS, and that the signal at these sidebands can be optimized by engineering the shape of the MNP.

  4. Fluorescence Resonance Energy Transfer Assay for High-Throughput Screening of ADAMTS1 Inhibitors

    Guanhua Du

    2011-12-01

    Full Text Available A disintegrin and metalloprotease with thrombospondin type I motifs-1 (ADAMTS1 plays a crucial role in inflammatory joint diseases and its inhibitors are potential candidates for anti-arthritis drugs. For the purposes of drug discovery, we reported the development and validation of fluorescence resonance energy transfer (FRET assay for high-throughput screening (HTS of the ADAMTS1 inhibitors. A FRET substrate was designed for a quantitative assay of ADAMTS1 activity and enzyme kinetics studies. The assay was developed into a 50-µL, 384-well assay format for high throughput screening of ADAMTS1 inhibitors with an overall Z’ factor of 0.89. ADAMTS1 inhibitors were screened against a diverse library of 40,960 total compounds with the established HTS system. Four structurally related hits, naturally occurring compounds, kuwanon P, kuwanon X, albafuran C and mulberrofuran J, extracted from the Chinese herb Morus alba L., were identified for further investigation. The results suggest that this FRET assay is an excellent tool, not only for measurement of ADAMTS1 activity but also for discovery of novel ADAMTS1 inhibitors with HTS.

  5. Monte Carlo treatment of resonance-radiation imprisonment in fluorescent lamps—revisited

    Anderson, James B.

    2016-12-01

    We reported in 1985 a Monte Carlo treatment of the imprisonment of the 253.7 nm resonance radiation from mercury in the mercury-argon discharge of fluorescent lamps. The calculated spectra of the emitted radiation were found in good agreement with measured spectra. The addition of the isotope mercury-196 to natural mercury was found, also in agreement with experiments, to increase lamp efficiency. In this paper we report the extension of the earlier work with increased accuracy, analysis of photon exit-time distributions, recycling of energy released in quenching, analysis of dynamic similarity for different lamp sizes, variation of Mrozowski transfer rates, prediction and analysis of the hyperfine ultra-violet spectra, and optimization of tailored mercury isotope mixtures for increased lamp efficiency. The spectra were found insensitive to the extent of quenching and recycling. The optimized mixtures were found to increase efficiencies by as much as 5% for several lamp configurations. Optimization without increasing the mercury-196 fraction was found to increase efficiencies by nearly 1% for several configurations.

  6. Monte Carlo treatment of resonance-radiation imprisonment in fluorescent lamps—revisited

    Anderson, James B

    2016-01-01

    We reported in 1985 a Monte Carlo treatment of the imprisonment of the 253.7 nm resonance radiation from mercury in the mercury–argon discharge of fluorescent lamps. The calculated spectra of the emitted radiation were found in good agreement with measured spectra. The addition of the isotope mercury-196 to natural mercury was found, also in agreement with experiments, to increase lamp efficiency. In this paper we report the extension of the earlier work with increased accuracy, analysis of photon exit-time distributions, recycling of energy released in quenching, analysis of dynamic similarity for different lamp sizes, variation of Mrozowski transfer rates, prediction and analysis of the hyperfine ultra-violet spectra, and optimization of tailored mercury isotope mixtures for increased lamp efficiency. The spectra were found insensitive to the extent of quenching and recycling. The optimized mixtures were found to increase efficiencies by as much as 5% for several lamp configurations. Optimization without increasing the mercury-196 fraction was found to increase efficiencies by nearly 1% for several configurations. (paper)

  7. Analytical use of multi-protein Fluorescence Resonance Energy Transfer to demonstrate membrane-facilitated interactions within cytokine receptor complexes.

    Krause, Christopher D; Izotova, Lara S; Pestka, Sidney

    2013-10-01

    Experiments measuring Fluorescence Resonance Energy Transfer (FRET) between cytokine receptor chains and their associated proteins led to hypotheses describing their organization in intact cells. These interactions occur within a larger protein complex or within a given nano-environment. To illustrate this complexity empirically, we developed a protocol to analyze FRET among more than two fluorescent proteins (multi-FRET). In multi-FRET, we model FRET among more than two fluorophores as the sum of all possible pairwise interactions within the complex. We validated our assumption by demonstrating that FRET among pairs within a fluorescent triplet resembled FRET between each pair measured in the absence of the third fluorophore. FRET between two receptor chains increases with increasing FRET between the ligand-binding chain (e.g., IFN-γR1, IL-10R1 and IFN-λR1) and an acylated fluorescent protein that preferentially resides within subsections of the plasma membrane. The interaction of IL-10R2 with IFN-λR1 or IL-10R1 results in decreased FRET between IL-10R2 and the acylated fluorescent protein. Finally, we analyzed FRET among four fluorescent proteins to demonstrate that as FRET between IFN-γR1 and IFN-γR2 or between IFN-αR1 and IFN-αR2c increases, FRET among other pairs of proteins changes within each complex. Copyright © 2013 Elsevier Ltd. All rights reserved.

  8. A lysosome-targetable turn-on fluorescent probe for the detection of thiols in living cells based on a 1,8-naphthalimide derivative

    Liang, Beibei; Wang, Baiyan; Ma, Qiujuan; Xie, Caixia; Li, Xian; Wang, Suiping

    2018-03-01

    Biological thiols, like cysteine (Cys), homocysteine (Hcy) and glutathione (GSH), play crucial roles in biological systems and in lysosomal processes. Highly selective probes for detecting biological thiols in lysomes of living cells are rare. In this work, a lysosome-targetable turn-on fluorescent probe for the detection of thiols in living cells was designed and synthesized based on a 1,8-naphthalimide derivative. The probe has a 4-(2-aminoethyl)morpholine unit as a lysosome-targetable group and an acrylate group as the thiol recognition unit as well as a fluorescence quencher. In the absence of biothiols, the probe displayed weak fluorescence due to the photoinduced electron transfer (PET) process. Upon the addition of biothiols, the probe exhibited an enhanced fluorescence emission centered at 550 nm due to cleavage of the acrylate moiety. The probe had high selectivity toward biothiols. Moreover, the probe features fast response time, excitation in the visible region and ability of working in a wide pH range. The linear response range covers a concentration range of Cys from 1.5 × 10- 7 to 1.0 × 10- 5 mol·L- 1 and the detection limit is 6.9 × 10- 8 mol·L- 1 for Cys. The probe has been successfully applied to the confocal imaging of biothiols in lysosomes of A549 cells with low cell toxicity. Furthermore, the method was successfully applied to the determination of thiols in a complex multicomponent mixture such as human serum, which suggests our proposed method has great potential for diagnostic purposes. All of such good properties prove it can be used to monitor biothiols in lysosomes of living cells and to be a good fluorescent probe for the selective detection of thiols.

  9. Detection of constitutive heterodimerization of the integrin Mac-1 subunits by fluorescence resonance energy transfer in living cells

    Fu Guo; Yang Huayan; Wang Chen; Zhang Feng; You Zhendong; Wang Guiying; He Cheng; Chen Yizhang; Xu Zhihan

    2006-01-01

    Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to β 2 subfamily of integrins that mediate important cell-cell and cell-extracellular matrix interactions. Biochemical studies have indicated that Mac-1 is a constitutive heterodimer in vitro. Here, we detected the heterodimerization of Mac-1 subunits in living cells by means of two fluorescence resonance energy transfer (FRET) techniques (fluorescence microscopy and fluorescence spectroscopy) and our results demonstrated that there is constitutive heterodimerization of the Mac-1 subunits and this constitutive heterodimerization of the Mac-1 subunits is cell-type independent. Through FRET imaging, we found that heterodimers of Mac-1 mainly localized in plasma membrane, perinuclear, and Golgi area in living cells. Furthermore, through analysis of the estimated physical distances between cyan fluorescent protein (CFP) and yellow fluorescent protein (YFP) fused to Mac-1 subunits, we suggested that the conformation of Mac-1 subunits is not affected by the fusion of CFP or YFP and inferred that Mac-1 subunits take different conformation when expressed in Chinese hamster ovary (CHO) and human embryonic kidney (HEK) 293T cells, respectively

  10. Palladium Nanoparticles-Based Fluorescence Resonance Energy Transfer Aptasensor for Highly Sensitive Detection of Aflatoxin M₁ in Milk.

    Li, Hui; Yang, Daibin; Li, Peiwu; Zhang, Qi; Zhang, Wen; Ding, Xiaoxia; Mao, Jin; Wu, Jing

    2017-10-13

    A highly sensitive aptasensor for aflatoxin M₁ (AFM₁) detection was constructed based on fluorescence resonance energy transfer (FRET) between 5-carboxyfluorescein (FAM) and palladium nanoparticles (PdNPs). PdNPs (33 nm) were synthesized through a seed-mediated growth method and exhibited broad and strong absorption in the whole ultraviolet-visible (UV-Vis) range. The strong coordination interaction between nitrogen functional groups of the AFM₁ aptamer and PdNPs brought FAM and PdNPs in close proximity, which resulted in the fluorescence quenching of FAM to a maximum extent of 95%. The non-specific fluorescence quenching caused by PdNPs towards fluorescein was negligible. After the introduction of AFM₁ into the FAM-AFM₁ aptamer-PdNPs FRET system, the AFM₁ aptamer preferentially combined with AFM₁ accompanied by conformational change, which greatly weakened the coordination interaction between the AFM₁ aptamer and PdNPs. Thus, fluorescence recovery of FAM was observed and a linear relationship between the fluorescence recovery and the concentration of AFM₁ was obtained in the range of 5-150 pg/mL in aqueous buffer with the detection limit of 1.5 pg/mL. AFM₁ detection was also realized in milk samples with a linear detection range from 6 pg/mL to 150 pg/mL. The highly sensitive FRET aptasensor with simple configuration shows promising prospect in detecting a variety of food contaminants.

  11. Detection of influenza A virus based on fluorescence resonance energy transfer from quantum dots to carbon nanotubes

    Tian Junping [Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian 116024 (China); Zhao Huimin, E-mail: zhaohuim@dlut.edu.cn [Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian 116024 (China); Liu Meng; Chen Yaqiong; Quan Xie [Key Laboratory of Industrial Ecology and Environmental Engineering (Ministry of Education, China), School of Environmental Science and Technology, Dalian University of Technology, Dalian 116024 (China)

    2012-04-20

    Highlights: Black-Right-Pointing-Pointer The quantum dots-ssDNA probe was designed for the determination of virus DNA. Black-Right-Pointing-Pointer The fluorescence of quantum dots was effectively quenched by carbon nanotubes. Black-Right-Pointing-Pointer The addition of target H5N1 DNA restored the quenched fluorescence of quantum dots. Black-Right-Pointing-Pointer The proposed method exhibited high sensitivity and good selectivity for H5N1 DNA. - Abstract: In this paper, a simple and sensitive approach for H5N1 DNA detection was described based on the fluorescence resonance energy transfer (FRET) from quantum dots (QDs) to carbon nanotubes (CNTs) in a QDs-ssDNA/oxCNTs system, in which the QDs (CdTe) modified with ssDNA were used as donors. In the initial stage, with the strong interaction between ssDNA and oxCNTs, QDs fluorescence was effectively quenched. Upon the recognition of the target, the effective competitive bindings of it to QDs-ssDNA occurred, which decreased the interactions between the QDs-ssDNA and oxCNTs, leading to the recovery of the QDs fluorescence. The recovered fluorescence of QDs was linearly proportional to the concentration of the target in the range of 0.01-20 {mu}M with a detection limit of 9.39 nM. Moreover, even a single-base mismatched target with the same concentration of target DNA can only recover a limited low fluorescence of QDs, illustrating the good anti-interference performance of this QDs-ssDNA/oxCNTs system. This FRET platform in the QDs-ssDNA/oxCNTs system was facilitated to the simple, sensitive and quantitative detection of virus nucleic acids and could have a wide range of applications in molecular diagnosis.

  12. Significantly improving nuclear resonance fluorescence non-destructive assay by using the integral resonance transmission method and photofission

    Angell, Christopher T.; Hayakawa, Takehito; Shizuma, Toshiyuki; Hajima, Ryoichi

    2013-01-01

    Non-destructive assay (NDA) of 239 Pu in spent nuclear fuel or melted fuel using a γ-ray beam is possible using self absorption and the integral resonance transmission method. The method uses nuclear resonance absorption where resonances in 239 Pu remove photons from the beam, and the selective absorption is detected by measuring the decrease in scattering in a witness target placed in the beam after the fuel, consisting of the isotope of interest, namely 239 Pu. The method is isotope specific, and can use photofission or scattered γ-rays to assay the 239 Pu. It overcomes several problems related to NDA of melted fuel, including the radioactivity of the fuel, and the unknown composition and geometry. This talk will explain the general method, and how photofission can be used to assay specific isotopes, and present example calculations. (author)

  13. Multifunctional PHPMA-Derived Polymer for Ratiometric pH Sensing, Fluorescence Imaging, and Magnetic Resonance Imaging.

    Su, Fengyu; Agarwal, Shubhangi; Pan, Tingting; Qiao, Yuan; Zhang, Liqiang; Shi, Zhengwei; Kong, Xiangxing; Day, Kevin; Chen, Meiwan; Meldrum, Deirdre; Kodibagkar, Vikram D; Tian, Yanqing

    2018-01-17

    In this paper, we report synthesis and characterization of a novel multimodality (MRI/fluorescence) probe for pH sensing and imaging. A multifunctional polymer was derived from poly(N-(2-hydroxypropyl)methacrylamide) (PHPMA) and integrated with a naphthalimide-based-ratiometric fluorescence probe and a gadolinium-1,4,7,10-tetraazacyclododecane-1,4,7,10-tetraacetic acid complex (Gd-DOTA complex). The polymer was characterized using UV-vis absorption spectrophotometry, fluorescence spectrofluorophotometry, magnetic resonance imaging (MRI), and confocal microscopy for optical and MRI-based pH sensing and cellular imaging. In vitro labeling of macrophage J774 and esophageal CP-A cell lines shows the polymer's ability to be internalized in the cells. The transverse relaxation time (T 2 ) of the polymer was observed to be pH-dependent, whereas the spin-lattice relaxation time (T 1 ) was not. The pH probe in the polymer shows a strong fluorescence-based ratiometric pH response with emission window changes, exhibiting blue emission under acidic conditions and green emission under basic conditions, respectively. This study provides new materials with multimodalities for pH sensing and imaging.

  14. Dynamic Measurement of Tumor Vascular Permeability and Perfusion using a Hybrid System for Simultaneous Magnetic Resonance and Fluorescence Imaging.

    Ren, Wuwei; Elmer, Andreas; Buehlmann, David; Augath, Mark-Aurel; Vats, Divya; Ripoll, Jorge; Rudin, Markus

    2016-04-01

    Assessing tumor vascular features including permeability and perfusion is essential for diagnostic and therapeutic purposes. The aim of this study was to compare fluorescence and magnetic resonance imaging (MRI)-based vascular readouts in subcutaneously implanted tumors in mice by simultaneous dynamic measurement of tracer uptake using a hybrid fluorescence molecular tomography (FMT)/MRI system. Vascular permeability was measured using a mixture of extravascular imaging agents, GdDOTA and the dye Cy5.5, and perfusion using a mixture of intravascular agents, Endorem and a fluorescent probe (Angiosense). Dynamic fluorescence reflectance imaging (dFRI) was integrated into the hybrid system for high temporal resolution. Excellent correspondence between uptake curves of Cy5.5/GdDOTA and Endorem/Angiosense has been found with correlation coefficients R > 0.98. The two modalities revealed good agreement regarding permeability coefficients and centers-of-gravity of the imaging agent distribution. The FMT/dFRI protocol presented is able to accurately map physiological processes and poses an attractive alternative to MRI for characterizing tumor neoangiogenesis.

  15. Graphene oxide based fluorescence resonance energy transfer and loop-mediated isothermal amplification for white spot syndrome virus detection.

    Waiwijit, U; Phokaratkul, D; Kampeera, J; Lomas, T; Wisitsoraat, A; Kiatpathomchai, W; Tuantranont, A

    2015-10-20

    Graphene oxide (GO) is attractived for biological or medical applications due to its unique electrical, physical, optical and biological properties. In particular, GO can adsorb DNA via π-π stacking or non-covalent interactions, leading to fluorescence quenching phenomenon applicable for bio-molecular detection. In this work, a new method for white spot syndrome virus (WSSV)-DNA detection is developed based on loop-mediated isothermal amplification (LAMP) combined with fluorescence resonance energy transfer (FRET) between GO and fluorescein isothiocyanate-labeled probe (FITC-probe). The fluorescence quenching efficiency of FITC-probe was found to increase with increasing GO concentration and reached 98.7% at a GO concentration of 50 μg/ml. The fluorescence intensity of FITC-probe was recovered after hybridization with WSSV LAMP product with an optimal hybridization time of 10 min and increased accordingly with increasing amount of LAMP products. The detection limit was estimated to be as low as 10 copies of WSSV plasmid DNA or 0.6 fg of the total DNA extracted from shrimp infected with WSSV. In addition, no cross reaction was observed with other common shrimp viral pathogens. Therefore, the GO-FRET-LAMP technique is promising for fast, sensitive and specific detection of DNAs. Copyright © 2015 Elsevier B.V. All rights reserved.

  16. A dark green fluorescent protein as an acceptor for measurement of Förster resonance energy transfer.

    Murakoshi, Hideji; Shibata, Akihiro C E; Nakahata, Yoshihisa; Nabekura, Junichi

    2015-10-15

    Measurement of Förster resonance energy transfer by fluorescence lifetime imaging microscopy (FLIM-FRET) is a powerful method for visualization of intracellular signaling activities such as protein-protein interactions and conformational changes of proteins. Here, we developed a dark green fluorescent protein (ShadowG) that can serve as an acceptor for FLIM-FRET. ShadowG is spectrally similar to monomeric enhanced green fluorescent protein (mEGFP) and has a 120-fold smaller quantum yield. When FRET from mEGFP to ShadowG was measured using an mEGFP-ShadowG tandem construct with 2-photon FLIM-FRET, we observed a strong FRET signal with low cell-to-cell variability. Furthermore, ShadowG was applied to a single-molecule FRET sensor to monitor a conformational change of CaMKII and of the light oxygen voltage (LOV) domain in HeLa cells. These sensors showed reduced cell-to-cell variability of both the basal fluorescence lifetime and response signal. In contrast to mCherry- or dark-YFP-based sensors, our sensor allowed for precise measurement of individual cell responses. When ShadowG was applied to a separate-type Ras FRET sensor, it showed a greater response signal than did the mCherry-based sensor. Furthermore, Ras activation and translocation of its effector ERK2 into the nucleus could be observed simultaneously. Thus, ShadowG is a promising FLIM-FRET acceptor.

  17. Improving the spectral analysis of Fluorescence Resonance Energy Transfer in live cells: application to interferon receptors and Janus kinases.

    Krause, Christopher D; Digioia, Gina; Izotova, Lara S; Pestka, Sidney

    2013-10-01

    The observed Fluorescence Resonance Energy Transfer (FRET) between fluorescently labeled proteins varies in cells. To understand how this variation affects our interpretation of how proteins interact in cells, we developed a protocol that mathematically separates donor-independent and donor-dependent excitations of acceptor, determines the electromagnetic interaction of donors and acceptors, and quantifies the efficiency of the interaction of donors and acceptors. By analyzing large populations of cells, we found that misbalanced or insufficient expression of acceptor or donor as well as their inefficient or reversible interaction influenced FRET efficiency in vivo. Use of red-shifted donors and acceptors gave spectra with less endogenous fluorescence but produced lower FRET efficiency, possibly caused by reduced quenching of red-shifted fluorophores in cells. Additionally, cryptic interactions between jellyfish FPs artefactually increased the apparent FRET efficiency. Our protocol can distinguish specific and nonspecific protein interactions even within highly constrained environments as plasma membranes. Overall, accurate FRET estimations in cells or within complex environments can be obtained by a combination of proper data analysis, study of sufficient numbers of cells, and use of properly empirically developed fluorescent proteins. Copyright © 2013 Elsevier Ltd. All rights reserved.

  18. Spectrophotometry near the atmospheric cutoff of the strongest Bowen resonance fluorescence lines of O III in two planetary nebulae

    O'Dell, C. R.; Opal, Chet B.

    1989-01-01

    Spectrophotometric results are presented for the stronger, well-resolved Bowen O III resonance fluorescence emission lines in the planetary nebulae 7027 and NGC 7662 down to and including the intrinsically strong line at 3133 A. These data are combined with results from the IUE atlas of spectra and similar results for the longer wavelength lines by Likkel and Aller (1986) to give the first full coverage of the Bowen lines. Good agreement is found with fluorescence theory for the primary cascade lines, except for the Likkel and Aller results. The efficiency of conversion of the exciting He II Ly-alpha into O III lines is determined, and values comparable to other planetary nebulae are found.

  19. A dark-field microscope for background-free detection of resonance fluorescence from single semiconductor quantum dots operating in a set-and-forget mode.

    Kuhlmann, Andreas V; Houel, Julien; Brunner, Daniel; Ludwig, Arne; Reuter, Dirk; Wieck, Andreas D; Warburton, Richard J

    2013-07-01

    Optically active quantum dots, for instance self-assembled InGaAs quantum dots, are potentially excellent single photon sources. The fidelity of the single photons is much improved using resonant rather than non-resonant excitation. With resonant excitation, the challenge is to distinguish between resonance fluorescence and scattered laser light. We have met this challenge by creating a polarization-based dark-field microscope to measure the resonance fluorescence from a single quantum dot at low temperature. We achieve a suppression of the scattered laser exceeding a factor of 10(7) and background-free detection of resonance fluorescence. The same optical setup operates over the entire quantum dot emission range (920-980 nm) and also in high magnetic fields. The major development is the outstanding long-term stability: once the dark-field point has been established, the microscope operates for days without alignment. The mechanical and optical designs of the microscope are presented, as well as exemplary resonance fluorescence spectroscopy results on individual quantum dots to underline the microscope's excellent performance.

  20. A dark-field microscope for background-free detection of resonance fluorescence from single semiconductor quantum dots operating in a set-and-forget mode

    Kuhlmann, Andreas V.; Houel, Julien; Warburton, Richard J.; Brunner, Daniel; Ludwig, Arne; Reuter, Dirk; Wieck, Andreas D.

    2013-01-01

    Optically active quantum dots, for instance self-assembled InGaAs quantum dots, are potentially excellent single photon sources. The fidelity of the single photons is much improved using resonant rather than non-resonant excitation. With resonant excitation, the challenge is to distinguish between resonance fluorescence and scattered laser light. We have met this challenge by creating a polarization-based dark-field microscope to measure the resonance fluorescence from a single quantum dot at low temperature. We achieve a suppression of the scattered laser exceeding a factor of 10 7 and background-free detection of resonance fluorescence. The same optical setup operates over the entire quantum dot emission range (920–980 nm) and also in high magnetic fields. The major development is the outstanding long-term stability: once the dark-field point has been established, the microscope operates for days without alignment. The mechanical and optical designs of the microscope are presented, as well as exemplary resonance fluorescence spectroscopy results on individual quantum dots to underline the microscope's excellent performance

  1. Measurement of a heavy-hole hyperfine interaction in InGaAs quantum dots using resonance fluorescence.

    Fallahi, P; Yilmaz, S T; Imamoğlu, A

    2010-12-17

    We measure the strength and the sign of hyperfine interaction of a heavy hole with nuclear spins in single self-assembled quantum dots. Our experiments utilize the locking of a quantum dot resonance to an incident laser frequency to generate nuclear spin polarization. By monitoring the resulting Overhauser shift of optical transitions that are split either by electron or exciton Zeeman energy with respect to the locked transition using resonance fluorescence, we find that the ratio of the heavy-hole and electron hyperfine interactions is -0.09 ± 0.02 in three quantum dots. Since hyperfine interactions constitute the principal decoherence source for spin qubits, we expect our results to be important for efforts aimed at using heavy-hole spins in quantum information processing.

  2. Characterization and inhibition of norovirus proteases of genogroups I and II using a fluorescence resonance energy transfer assay

    Chang, Kyeong-Ok; Takahashi, Daisuke; Prakash, Om; Kim, Yunjeong

    2012-01-01

    Noroviruses are the major cause of food- or water-borne gastroenteritis outbreaks in humans. The norovirus protease that cleaves a large viral polyprotein to nonstructural proteins is essential for virus replication and an attractive target for antiviral drug development. Noroviruses show high genetic diversity with at least five genogroups, GI–GV, of which GI and GII are responsible for the majority of norovirus infections in humans. We cloned and expressed proteases of Norwalk virus (GI) and MD145 virus (GII) and characterized the enzymatic activities with fluorescence resonance energy transfer substrates. We demonstrated that the GI and GII proteases cleaved the substrates derived from the naturally occurring cleavage site in the open reading frame (ORF) 1 of G1 norovirus with similar efficiency, and that enzymatic activity of both proteases was inhibited by commercial protease inhibitors including chymostatin. The interaction of chymostatin to Norwalk virus protease was validated by nuclear magnetic resonance (NMR) spectroscopy.

  3. Intensity and pressure dependence of resonance fluorescence of OH induced by a tunable UV laser

    Killinger, D. K.; Wang, C. C.; Hanabusa, M.

    1976-01-01

    The intensity and pressure dependence of the fluorescence spectrum of OH in the presence of N2 and H2O molecules was studied. Saturation of the absorption transition was observed at low pressures, and the corresponding fluorescence signal was found to vary as the square root of the exciting intensity. This observed dependence agreed with the predicted dependence which took into account the presence of laser modes in the spectrum of the exciting radiation. With full laser power incident, a saturation parameter as high as 3 x 10 to the 5th was observed. The fluorescence spectrum was found to peak at 3145 and at 3090 A, with the relative peak intensities dependent upon gas pressures and upon the particular rotational electronic transition used for excitation. It is concluded that vibrational relaxation of the electronically excited OH due to water vapor in the system plays a dominant role in determining the observed fluorescence spectrum.

  4. Full genotyping of a highly polymorphic human gene trait by time-resolved fluorescence resonance energy transfer.

    Edoardo Totè

    Full Text Available The ability of detecting the subtle variations occurring, among different individuals, within specific DNA sequences encompassed in highly polymorphic genes discloses new applications in genomics and diagnostics. DQB1 is a gene of the HLA-II DQ locus of the Human Leukocyte Antigens (HLA system. The polymorphisms of the trait of the DQB1 gene including codons 52-57 modulate the susceptibility to a number of severe pathologies. Moreover, the donor-receiver tissue compatibility in bone marrow transplantations is routinely assessed through crossed genotyping of DQB and DQA. For the above reasons, the development of rapid, reliable and cost-effective typing technologies of DQB1 in general, and more specifically of the codons 52-57, is a relevant although challenging task. Quantitative assessment of the fluorescence resonance energy transfer (FRET efficiency between chromophores labelling the opposite ends of gene-specific oligonucleotide probes has proven to be a powerful tool to type DNA polymorphisms with single-nucleotide resolution. The FRET efficiency can be most conveniently quantified by applying a time-resolved fluorescence analysis methodology, i.e. time-correlated single-photon counting, which allows working on very diluted template specimens and in the presence of fluorescent contaminants. Here we present a full in-vitro characterization of the fluorescence responses of two probes when hybridized to oligonucleotide mixtures mimicking all the possible genotypes of the codons 52-57 trait of DQB1 (8 homozygous and 28 heterozygous. We show that each genotype can be effectively tagged by the combination of the fluorescence decay constants extrapolated from the data obtained with such probes.

  5. Blinking fluorescence of single donor-acceptor pairs: important role of "dark'' states in resonance energy transfer via singlet levels.

    Osad'ko, I S; Shchukina, A L

    2012-06-01

    The influence of triplet levels on Förster resonance energy transfer via singlet levels in donor-acceptor (D-A) pairs is studied. Four types of D-A pair are considered: (i) two-level donor and two-level acceptor, (ii) three-level donor and two-level acceptor, (iii) two-level donor and three-level acceptor, and (iv) three-level donor and three-level acceptor. If singlet-triplet transitions in a three-level acceptor molecule are ineffective, the energy transfer efficiency E=I_{A}/(I_{A}+I_{D}), where I_{D} and I_{A} are the average intensities of donor and acceptor fluorescence, can be described by the simple theoretical equation E(F)=FT_{D}/(1+FT_{D}). Here F is the rate of energy transfer, and T_{D} is the donor fluorescence lifetime. In accordance with the last equation, 100% of the donor electronic energy can be transferred to an acceptor molecule at FT_{D}≫1. However, if singlet-triplet transitions in a three-level acceptor molecule are effective, the energy transfer efficiency is described by another theoretical equation, E(F)=F[over ¯](F)T_{D}/[1+F[over ¯](F)T_{D}]. Here F[over ¯](F) is a function of F depending on singlet-triplet transitions in both donor and acceptor molecules. Expressions for the functions F[over ¯](F) are derived. In this case the energy transfer efficiency will be far from 100% even at FT_{D}≫1. The character of the intensity fluctuations of donor and acceptor fluorescence indicates which of the two equations for E(F) should be used to find the value of the rate F. Therefore, random time instants of photon emission in both donor and acceptor fluorescence are calculated by the Monte Carlo method for all four types of D-A pair. Theoretical expressions for start-stop correlators (waiting time distributions) in donor and acceptor fluorescence are derived. The probabilities w_{N}^{D}(t) and w_{N}^{A}(t) of finding N photons of donor and acceptor fluorescence in the time interval t are calculated for various values of the energy

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

    Hsu, Y.-Y.; Liu, Y.-N.; Wang Wenyen; Kao, Fu-Jen; Kung, S.-H.

    2007-01-01

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

  7. Method of using a nuclear magnetic resonance spectroscopy standard. [SO/sub 2/ in gases by fluorescence

    Spicer, L.D.; Bennett, D.W.; Davis, J.F.

    1983-05-09

    (CH/sub 3/)/sub 3/SiNSO is produced by the reaction of ((CH/sub 3/)/sub 3/SI)/sub 2/NH with SO/sub 2/. Also produced in the reaction are ((CH/sub 3/)/sub 3/Si)/sub 2/O and a new solid compound (NH/sub 4/)((CH/sub 3/)/sub 3/SiOSO/sub 2/). Both (CH/sub 3/)/sub 3/SiNSO and (NH/sub 4/)((CH/sub 3/)/sub 3/SiOSO/sub 2/) have fluorescent properties. The reaction of the subject invention is used in a method of measuring the concentration of SO/sub 2/ pollutants in gases. By the method, a sample of gas is bubbled through a solution of ((CH/sub 3/)/sub 3/Si)/sub 2/NH, whereby any SO/sub 2/ present in the gas will react to produce the two fluorescent products. The measured fluorescence of these products can then be used to calculate the concentration of SO/sub 2/ in the original gas sample. The solid product (NH/sub 4/)((CH/sub 3/)/sub 3/SiOSO/sub 2/) may be used as a standard in solid state NMR spectroscopy, wherein the resonance peaks of either /sup 1/H, /sup 13/C, /sup 15/N, or /sup 29/Si may be used as a reference.

  8. Diphenylacrylonitrile-connected BODIPY dyes: fluorescence enhancement based on dark and AIE resonance energy transfer.

    Lin, Liangbin; Lin, Xiaoru; Guo, Hongyu; Yang, Fafu

    2017-07-19

    This study focuses on the construction of novel diphenylacrylonitrile-connected BODIPY dyes with high fluorescence in both solution and an aggregated state by combining DRET and FRET processes in a single donor-acceptor system. The first BODIPY derivatives with one, two, or three AIE-active diphenylacrylonitrile groups were designed and synthesized in moderate yields. Strong fluorescence emissions were observed in the THF solution under excitation at the absorption wavelength of non-emissive diphenylacrylonitrile chromophores, implying the existence of the DRET process between the dark diphenylacrylonitrile donor and the emissive BODIPY acceptor. In the THF/H 2 O solution, the fluorescence intensity of the novel BODIPY derivatives gradually increased under excitation at the absorption wavelength of diphenylacrylonitrile chromophores, suggesting a FRET process between diphenylacrylonitrile and BODIPY moieties. A greater number of diphenylacrylonitrile units led to higher energy-transfer efficiencies. The pseudo-Stokes shift for both DRET and FRET processes was as large as 190 nm.

  9. Fluorescent strategy based on cationic conjugated polymer fluorescence resonance energy transfer for the quantification of 5-(hydroxymethyl)cytosine in genomic DNA.

    Hong, Tingting; Wang, Tianlu; Guo, Pu; Xing, Xiwen; Ding, Fei; Chen, Yuqi; Wu, Jinjun; Ma, Jingwei; Wu, Fan; Zhou, Xiang

    2013-11-19

    DNA methylation is dynamically reprogrammed during early embryonic development in mammals. It can be explained partially by the discovery of 5-(hydroxymethyl)cytosine (5-hmC), 5-formylcytosine (5-fC), and 5-carboxylcytosine (5-caC), which are identified as key players involved in both active and passive demethylation pathways. As one of the ten-eleven translocation oxidation products, 5-hmC was found relatively abundant in neuron cells and embryonic stem cells. Herein we report a new method for 5-hmC quantification in genomic DNA based on CCP-FRET (cationic conjugated polymers act as the energy donor and induce fluorescence resonance energy transfer) assay combined with KRuO4 oxidation. 5-hmC in genomic DNA can be selectively transformed into 5-fC by the oxidation of KRuO4 and then labeled with hydroxylamine-BODIPY (BODIPY = 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene) fluorophore through the reaction between 5-fC and hydroxylamine-BODIPY. After the fluorescently labeled DNA was captured by CCP through electrostatic interactions, a significant FRET between CCP and hydroxylamine-BODIPY fluorophore was observed. This CCP-FRET-based assay benefits from light-harvesting, large Stokes shift, and optical signal amplification properties of the CCP. Furthermore, this CCP-FRET-based assay was quite successfully demonstrated for the 5-hmC quantification in three types of cells (mESc, HeLa, HEK 293T), providing a much more convenient choice for 5-hmC quantification in genomic DNA.

  10. Stepwise synthesis of cubic Au-AgCdS core-shell nanostructures with tunable plasmon resonances and fluorescence.

    Liu, Xiao-Li; Liang, Shan; Nan, Fan; Pan, Yue-Yue; Shi, Jun-Jun; Zhou, Li; Jia, Shuang-Feng; Wang, Jian-Bo; Yu, Xue-Feng; Wang, Qu-Quan

    2013-10-21

    Cubic Au-AgCdS core-shell nanostructures were synthesized through cation exchange method assisted by tributylphosphine (TBP) as a phase-transfer agent. Among intermediate products, Au-Ag core-shell nanocubes exhibited many high-order plasmon resonance modes related to the special cubic shape, and these plasmon bands red-shifted along with the increasing of particle size. The plasmon band of Au core first red-shifted and broadened at the step of Au-Ag₂S and then blue-shifted and narrowed at the step of Au-AgCdS. Since TBP was very crucial for the efficient conversion from Ag₂S to CdS, we found that both absorption and fluorescence of the final products could be controlled by TBP.

  11. Using biharmonic laser pumping for preparation of pure and entangled multiexciton states in clusters of resonantly interacting fluorescent centres

    Basieva, I.T.; Basiev, T.T.; Dietler, G.; Pukhov, K.K.; Sekatskii, S.K.

    2007-01-01

    Use of a biharmonic laser pumping for preparation of pure and entangled multiexciton states in dimers and tetramers of resonantly interacting fluorescent particles is analysed. Special emphasis is given to the preparation of all possible pure exciton states and their maximally entangled Bell states. The general results are illustrated using as an example the pair and quartet centres of neodymium ions in calcium fluoride (M- and N-centres), where all necessary experimental information concerning the interactions and decoherence is available, and experimental preparation of Bell vacuum-single exciton and vacuum-biexciton states has been recently demonstrated. These results can be easily rescaled for the cases of quantum dots and dye molecules. Numerical results are compared with the analytical results obtained for a particular case of the biharmonic excitation of dimers. Excellent agreement between these approaches is demonstrated

  12. Time-resolved spectroscopy and fluorescence resonance energy transfer in the study of excimer laser damage of chromatin

    Radu, L. [Department of Molecular Genetics and Radiobiology, Babes National Institute, Bucharest (Romania)], E-mail: lilianajradu@yahoo.fr; Mihailescu, I. [Department of Lasers, Laser, Plasma and Radiation Physics Institute, Bucharest (Romania); Radu, S. [Department of Computer Science, Polytechnics University, Bucharest (Romania); Gazdaru, D. [Department of Biophysics, Bucharest University (Romania)

    2007-09-21

    The analysis of chromatin damage produced by a 248 nm excimer laser radiation, for doses of 0.3-3 MJ/m{sup 2} was carried out by time-resolved spectroscopy and fluorescence resonance energy transfer (FRET). The chromatin was extracted from a normal and a tumoral tissue of Wistar rats. The decrease with laser dose of the relative contribution of the excited state lifetimes of ethidium bromide (EtBr) bounded to chromatin constitutes an evidence of the reduction of chromatin deoxyribonucleic acid (DNA) double-strand structure. FRET was performed from dansyl chloride to acridine orange, both coupled to chromatin. The increase of the average distance between these ligands, under the action of laser radiation, reflects a loosening of the chromatin structure. The radiosensitivity of tumor tissue chromatin is higher than that of a normal tissue. The determination of the chromatin structure modification in an excimer laser field can be of interest in laser therapy.

  13. Time-resolved spectroscopy and fluorescence resonance energy transfer in the study of excimer laser damage of chromatin

    Radu, L.; Mihailescu, I.; Radu, S.; Gazdaru, D.

    2007-01-01

    The analysis of chromatin damage produced by a 248 nm excimer laser radiation, for doses of 0.3-3 MJ/m 2 was carried out by time-resolved spectroscopy and fluorescence resonance energy transfer (FRET). The chromatin was extracted from a normal and a tumoral tissue of Wistar rats. The decrease with laser dose of the relative contribution of the excited state lifetimes of ethidium bromide (EtBr) bounded to chromatin constitutes an evidence of the reduction of chromatin deoxyribonucleic acid (DNA) double-strand structure. FRET was performed from dansyl chloride to acridine orange, both coupled to chromatin. The increase of the average distance between these ligands, under the action of laser radiation, reflects a loosening of the chromatin structure. The radiosensitivity of tumor tissue chromatin is higher than that of a normal tissue. The determination of the chromatin structure modification in an excimer laser field can be of interest in laser therapy

  14. A comparison of rapid-scanning X-ray fluorescence mapping and magnetic resonance imaging to localize brain iron distribution

    McCrea, Richard P.E.; Harder, Sheri L.; Martin, Melanie; Buist, Richard; Nichol, Helen

    2008-01-01

    The clinical diagnosis of many neurodegenerative disorders relies primarily or exclusively on observed behaviors rather than measurable physical tests. One of the hallmarks of Alzheimer disease (AD) is the presence of amyloid-containing plaques associated with deposits of iron, copper and/or zinc. Work in other laboratories has shown that iron-rich plaques can be seen in the mouse brain in vivo with magnetic resonance imaging (MRI) using a high-field strength magnet but this iron cannot be visualized in humans using clinical magnets. To improve the interpretation of MRI, we correlated iron accumulation visualized by X-ray fluorescence spectroscopy, an element-specific technique with T1, T2, and susceptibility weighted MR (SWI) in a mouse model of AD. We show that SWI best shows areas of increased iron accumulation when compared to standard sequences

  15. Application of multivariate curve resolution for the study of folding processes of DNA monitored by fluorescence resonance energy transfer

    Kumar, Praveen; Kanchan, Kajal; Gargallo, Raimundo; Chowdhury, Shantanu

    2005-01-01

    The study described in the present article used fluorescence resonance energy transfer (FRET) to monitor the folding of a 31-mer cytosine-rich DNA segment, from the promoter region of the human c-myc oncogene. Spectroscopic FRET data recorded during experiments carried out in different experimental conditions were individually and simultaneously analyzed by multivariate curve resolution. The simultaneous analysis of several data matrices allowed the resolution of the system, removing most of the ambiguities related to factor analysis. From the results obtained, we report the evidence of the formation of two ordered conformations in acidic and neutral pH values, in addition to the disordered structure found at high temperatures. These ordered conformations could be related to cytosine-tetraplex structures showing different degrees of protonation in cytosine bases

  16. Optical bar code recognition of methyl salicylate (MES) for environmental monitoring using fluorescence resonance energy transfer (FRET) on thin films

    Smith, Clint; Tatineni, Balaji; Anderson, John; Tepper, Gary

    2006-10-01

    Fluorescence resonance energy transfer (FRET) is a process in which energy is transferred nonradiatively from one fluorophore (the donor) in an excited electron state to another, the chromophore (the acceptor). FRET is distinctive in its ability to reveal the presence of specific recognition of select targets such as the nerve agent stimulant Methyl Salicylate (MES) upon spectroscopic excitation. We introduce a surface imprinted and non-imprinted thin film that underwent AC-Electrospray ionization for donor-acceptor pair(s) bound to InGaP quantum dots and mesoporous silicate nanoparticles. The donor-acceptor pair used in this investigation included MES (donor) and 6-(fluorescein-5-(and-6)- carboxamido) hexanoic acid, succinimidyl ester bound to InGaP quantum dots (acceptor). MES was then investigated as a donor to various acceptor fluorophore: InGaP: mesoporous silicate nanoparticle layers.

  17. Imaging of activated caspase-3 in living cell by fluorescence resonance energy transfer during photosensitization-induced apoptosis

    Wu, Yunxia; Xing, Da; Chen, Qun; Tang, Yonghong

    2005-01-01

    Photodynamic therapy (PDT) is a novel and promising cancer treatment that employs a combination of a photosensitizing chemical and visible light, induces apoptosis in cell, and activation of caspase-3 is considered to be the final step in many apoptosis pathways. The changes of caspase-3 activation in cell during TNFα- and photodynamic therapy-induced apoptosis was measured by fluorescence resonance energy transfer (FRET) analysis. FRET probe consisting of fusions of an enhanced cyan fluorescent protein (ECFP), Venus and a linker peptide containing the caspase-3 cleavage sequence DEVD was utilized. Therefore, activated caspase-3 cleaved the linker peptide of FRET probe and disrupted the FRET signal. Human lung adenocarcinoma cell line (ASTC-a-1) were stably transfected with the plasmid (ECFP-DEVD-Venus) and then were treated by TNF-α and PDT, respectively. Experimental results indicated that caspase-3 activation resulted in cleavage of linker peptide and subsequent disruption of the FRET signal during TNFα- and photodynamic therapy-induced apoptosis, and that the activation of caspase-3 induced by photodynamic therapy was faster than that induce by TNF-α. The study supports that using FRET technique and different recombinant substrates as FRET probes could be used to detect the process of PDT-induced apoptosis and provide a new means to investigate apoptotic mechanism of PDT.

  18. Quantification of Material Fluorescence and Light Scattering Cross Sections Using Ratiometric Bandwidth-Varied Polarized Resonance Synchronous Spectroscopy.

    Xu, Joanna Xiuzhu; Hu, Juan; Zhang, Dongmao

    2018-05-25

    Presented herein is the ratiometric bandwidth-varied polarized resonance synchronous spectroscopy (BVPRS2) method for quantification of material optical activity spectra. These include the sample light absorption and scattering cross-section spectrum, the scattering depolarization spectrum, and the fluorescence emission cross-section and depolarization spectrum in the wavelength region where the sample both absorbs and emits. This ratiometric BVPRS2 spectroscopic method is a self-contained technique capable of quantitatively decoupling material fluorescence and light scattering signal contribution to its ratiometric BVPRS2 spectra through the linear curve-fitting of the ratiometric BVPRS2 signal as a function of the wavelength bandwidth used in the PRS2 measurements. Example applications of this new spectroscopic method are demonstrated with materials that can be approximated as pure scatterers, simultaneous photon absorbers/emitters, simultaneous photon absorbers/scatterers, and finally simultaneous photon absorbers/scatterers/emitters. Because the only instruments needed for this ratiometric BVPRS2 technique are the conventional UV-vis spectrophotometer and spectrofluorometer, this work should open doors for routine decomposition of material UV-vis extinction spectrum into its absorption and scattering component spectra. The methodology and insights provided in this work should be of broad significance to all chemical research that involves photon/matter interactions.

  19. Deciphering the fluorescence resonance energy transfer from denatured transport protein to anthracene 1,5 disulphonate in reverse micellar environment

    Singharoy, Dipti; Bhattacharya, Subhash Chandra

    2017-12-01

    Constrained environmental effect inside AOT reverse micellar media has been employed in this work to collect the information about energy transfer efficacy between sodium salt of anthracene 1,5 disulphonate (1,5-AS) with model transport proteins, bovine serum albumin (BSA), and human serum albumin (HSA). Steady state, time-resolved fluorescence and circular dichroism techniques have been used for this purpose and corresponding Fӧrster-type resonance energy transfer (FRET) from tryptophan residues to 1,5-AS indicates that 1,5-AS binds in the vicinity of the tryptophan residue (BSA and HSA) with equal strength. Indication of protein damage from fluorescence data and its confirmation has been measured from CD measurement. Molecular modeling study hereby plays a crucial role to predict the minimum energy docked conformation of the probe inside the protein environment. From the docked conformation the distance between 1,5-AS and tryptophan moiety of BSA/HSA has successfully explained the FRET possibility between them. A comparative modeling study between BSA and HSA with 1,5-AS assigning their binding site within specific amino acids plays a crucial role in support of the FRET study.

  20. Evaluation by fluorescence resonance energy transfer of the stability of nonviral gene delivery vectors under physiological conditions.

    Itaka, Keiji; Harada, Atsushi; Nakamura, Kozo; Kawaguchi, Hiroshi; Kataoka, Kazunori

    2002-01-01

    The stability in physiological medium of polyplex- and lipoplex-type nonviral gene vectors was evaluated by detecting the conformational change of complexed plasmid DNA (pDNA) labeled simultaneously with fluorescein (energy donor) and X-rhodamine (energy acceptor) through fluorescence resonance energy transfer (FRET). Upon mixing with cationic components, such as LipofectAMINE, poly(L-lysine), and poly(ethylene glycol)-poly(L-lysine) block copolymer (PEG-PLys), the fluorescence spectrum of doubly labeled pDNA underwent a drastic change due to the occurrence of FRET between the donor-acceptor pair on pDNA taking a globular conformation (condensed state) through complexation. The measurement was carried out also in the presence of 20% serum, under which conditions FRET from condensed pDNA was clearly monitored without interference from coexisting components in the medium, allowing evaluation of the condensed state of pDNA in nonviral gene vectors under physiological conditions. Serum addition immediately induced a sharp decrease in FRET for the LipofectAMINE/pDNA (lipoplex) system, which was consistent with the sharp decrease in the transfection efficiency of the lipoplex system in serum-containing medium. In contrast, the PEG-PLys/pDNA polyplex (polyion complex micelle) system maintained appreciable transfection efficiency even in serum-containing medium, and FRET efficiency remained constant for up to 12 h, indicating the high stability of the polyion complex micelle under physiological conditions.

  1. Lifetime-based optical sensor for high-level pCO2 detection employing fluorescence resonance energy transfer

    Bueltzingsloewen, Christoph von; McEvoy, Aisling K.; McDonagh, Colette; MacCraith, Brian D.

    2003-01-01

    An optical sensor for the measurement of high levels of carbon dioxide in gas phase has been developed. It is based on fluorescence resonance energy transfer (FRET) between a long-lifetime ruthenium polypyridyl complex and the pH-active disazo dye Sudan III. The donor luminophore and the acceptor dye are both immobilised in a hydrophobic silica sol-gel/ethyl cellulose hybrid matrix material. Tetraoctylammonium hydroxide (TOA-OH) is used as an internal buffering system. Fluorescence lifetime is measured in the frequency domain, using low-cost phase modulation measurement technology. The use of Sudan III as an acceptor dye has enabled the sensor to have a dynamic range up to 100% carbon dioxide. The sensor displays 11.2 deg. phase shift between the limit of detection (LOD) of 0.06 and 100% CO 2 with a resolution of better than 2%. The encapsulation in the silica/polymer hybrid material has provided the sensor with good mechanical and chemical stability. The effect of molecular oxygen, humidity and temperature on the sensor performance was studied in detail

  2. On-chip transduction of nucleic acid hybridization using spatial profiles of immobilized quantum dots and fluorescence resonance energy transfer.

    Tavares, Anthony J; Noor, M Omair; Vannoy, Charles H; Algar, W Russ; Krull, Ulrich J

    2012-01-03

    The glass surface of a glass-polydimethylsiloxane (PDMS) microfluidic channel was modified to develop a solid-phase assay for quantitative determination of nucleic acids. Electroosmotic flow (EOF) within channels was used to deliver and immobilize semiconductor quantum dots (QDs), and electrophoresis was used to decorate the QDs with oligonucleotide probe sequences. These processes took only minutes to complete. The QDs served as energy donors in fluorescence resonance energy transfer (FRET) for transduction of nucleic acid hybridization. Electrokinetic injection of fluorescent dye (Cy3) labeled oligonucleotide target into a microfluidic channel and subsequent hybridization (within minutes) provided the proximity for FRET, with emission from Cy3 being the analytical signal. The quantification of target concentration was achieved by measurement of the spatial length of coverage by target along a channel. Detection of femtomole quantities of target was possible with a dynamic range spanning an order of magnitude. The assay provided excellent resistance to nonspecific interactions of DNA. Further selectivity of the assay was achieved using 20% formamide, which allowed discrimination between a fully complementary target and a 3 base pair mismatch target at a contrast ratio of 4:1. © 2011 American Chemical Society

  3. A fluorescence detected magnetic resonance investigation of the carotenoid triplet states associated with Photosystem II of isolated spinach thylakoid membranes

    Santabarbara, S; Carbonera, D; Heathcote, P

    2005-01-01

    The carotenoid triplet populations associated with the fluorescence emission chlorophyll forms of Photosystem II have been investigated in isolated spinach thylakoid membranes by means of fluorescence detected magnetic resonance in zero field (FDMR). The spectra collected in the 680-690 nm emission range, have been fitted by a global analysis procedure. At least five different carotenoid triplet states coupled to the terminal emitting chlorophyll forms of PS II, peaking at 682 nm, 687 nm and 692 nm, have been characterised. The triplets associated with the outer antenna emission forms, at 682 nm, have zero field splitting parameters D = 0.0385 cm/sup -1/, E = 0.00367 cm/sup -1/; D = 0.0404 cm/sup -1/, E = 0.00379 cm/sup -1/ and D = 0.0386 cm/sup -1/, E = 0.00406 cm/sup -1/ which are very similar to those previously reported for the xanthophylls of the isolated LHC II complex. Therefore the FDMR spectra recorded in this work provide insights into the organisation of the LHC II complex in the unperturbed enviro...

  4. Efficient Fluorescence Resonance Energy Transfer between Quantum Dots and Gold Nanoparticles Based on Porous Silicon Photonic Crystal for DNA Detection.

    Zhang, Hongyan; Lv, Jie; Jia, Zhenhong

    2017-05-10

    A novel assembled biosensor was prepared for detecting 16S rRNA, a small-size persistent specific for Actinobacteria. The mechanism of the porous silicon (PS) photonic crystal biosensor is based on the fluorescence resonance energy transfer (FRET) between quantum dots (QDs) and gold nanoparticles (AuNPs) through DNA hybridization, where QDs act as an emission donor and AuNPs serve as a fluorescence quencher. Results showed that the photoluminescence (PL) intensity of PS photonic crystal was drastically increased when the QDs-conjugated probe DNA was adhered to the PS layer by surface modification using a standard cross-link chemistry method. The PL intensity of QDs was decreased when the addition of AuNPs-conjugated complementary 16S rRNA was dropped onto QDs-conjugated PS. Based on the analysis of different target DNA concentration, it was found that the decrease of the PL intensity showed a good linear relationship with complementary DNA concentration in a range from 0.25 to 10 μM, and the detection limit was 328.7 nM. Such an optical FRET biosensor functions on PS-based photonic crystal for DNA detection that differs from the traditional FRET, which is used only in liquid. This method will benefit the development of a new optical FRET label-free biosensor on Si substrate and has great potential in biochips based on integrated optical devices.

  5. Identification of weak autoionizing resonances observed through fluorescence from the satellite states of Ar{sup +}

    McLaughlin, K.W.; Yenen, O.; Samson, J.A.R. [Univ. of Nebraska, Lincoln, NE (United States)] [and others

    1997-04-01

    Photoionization accompanied by excitation of the residual ionic state violates an independent electron model since, according to QED, photons interact only with individual electrons. By allowing measurements at a threshold event with high resolution, the observation of the fluorescence from the decay of these excited states (satellite states) is a sensitive method in the study of electron-electron interactions, providing complementary information to photoelectron spectroscopy. In the measurements reported here, an atomic beam of argon has been photoionized with 34 to 39 eV synchrotron radiation at beamline 9.0.1 of the Advanced Light Source. This energy range encompasses the 3p{sup 4} [{sup 3}P] 4p {sup 4}P, {sup 2}P, and {sup 2}D as well as the [{sup 1}D]4p {sup 2}F satellite states of Ar{sup +}. By observing the fine-structure resolved fluorescence from these satellite states, new Rydberg series and extensions of previously known series have been resolved with an energy resolution of 3 meV. With the high photon flux available from the high resolution monochromator of beamline 9.0.1, even the weakly excited [{sup 3}P] 4p ({sup 2}S) ns,d autoionizing structure has been observed for the first time.

  6. Identification of weak autoionizing resonances observed through fluorescence from the satellite states of Ar+

    McLaughlin, K.W.; Yenen, O.; Samson, J.A.R.

    1997-01-01

    Photoionization accompanied by excitation of the residual ionic state violates an independent electron model since, according to QED, photons interact only with individual electrons. By allowing measurements at a threshold event with high resolution, the observation of the fluorescence from the decay of these excited states (satellite states) is a sensitive method in the study of electron-electron interactions, providing complementary information to photoelectron spectroscopy. In the measurements reported here, an atomic beam of argon has been photoionized with 34 to 39 eV synchrotron radiation at beamline 9.0.1 of the Advanced Light Source. This energy range encompasses the 3p 4 [ 3 P] 4p 4 P, 2 P, and 2 D as well as the [ 1 D]4p 2 F satellite states of Ar + . By observing the fine-structure resolved fluorescence from these satellite states, new Rydberg series and extensions of previously known series have been resolved with an energy resolution of 3 meV. With the high photon flux available from the high resolution monochromator of beamline 9.0.1, even the weakly excited [ 3 P] 4p ( 2 S) ns,d autoionizing structure has been observed for the first time

  7. Rapid creation of distant entanglement by multi-photon resonant fluorescence

    Cohen, Guy Z.; Sham, L. J.

    2014-03-01

    We study a simple, effective and robust method for entangling two separate stationary quantum dot spin qubits with high fidelity using multi-photon Gaussian state. The fluorescence signals from the two dots interfere at a beam splitter. The bosonic nature of photons leads, in analogy with the Hong-Ou-Mandel (HOM) effect, to selective pairing of photon holes (photon absences in the fluorescent signals). By the HOM effect, two photon holes with the same polarization end up at the same beam splitter output. As a result, two odd photon number detections at the outgoing beams, which must correspond to two photon holes with different polarizations, herald entanglement creation. The robustness of the Gaussian states is evidenced by the ability to compensate for photon absorption and noise by a moderate increase in the number of photons at the input. We calculate the entanglement generation rate in the ideal, non-ideal and near-ideal detector regimes and find substantial improvement over single-photon schemes in all three regimes. Fast and efficient spin-spin entanglement creation can form the basis for a scalable quantum dot quantum computing network. Our predictions can be tested using current experimental capabilities. This research was supported by the U.S. Army Research Office MURI award W911NF0910406, by NSF grant PHY-1104446 and by ARO (IARPA, W911NF-08-1-0487). The authors thank D. G. Steel for useful discussions.

  8. A new assay format for NF-kappaB based on a DNA triple helix and a fluorescence resonance energy transfer.

    Altevogt, Dominik; Hrenn, Andrea; Kern, Claudia; Clima, Lilia; Bannwarth, Willi; Merfort, Irmgard

    2009-10-07

    Herein we report a feasibility study for a new concept to detect DNA binding protein NF-kappaB based on a DNA triple helix formation in combination with a fluorescence resonance energy transfer (FRET). The new principle avoids expensive antibodies and radioactivity and might have implications for assays of other DNA binding proteins.

  9. Homogeneous competitive assay of ligand affinities based on quenching fluorescence of tyrosine/tryptophan residues in a protein via Főrster-resonance-energy-transfer

    Xie, Yanling; Yang, Xiaolan; Pu, Jun; Zhao, Yunsheng; Zhang, Ying; Xie, Guoming; Zheng, Jun; Yuan, Huidong; Liao, Fei

    2010-11-01

    A new homogeneous competitive assay of ligand affinities was proposed based on quenching the fluorescence of tryptophan/tyrosine residues in a protein via Főrster-resonance-energy-transfer using a fluorescent reference ligand as the acceptor. Under excitation around 280 nm, the fluorescence of a protein or a bound acceptor was monitored upon competitive binding against a nonfluorescent candidate ligand. Chemometrics for deriving the binding ratio of the acceptor with either fluorescence signal was discussed; the dissociation constant ( Kd) of a nonfluorescent candidate ligand was calculated from its concentration to displace 50% binding of the acceptor. N-biotinyl-N'-(1-naphthyl)-ethylenediamine (BNEDA) and N-biotinyl-N'-dansyl-ethylenediamine (BDEDA) were used as the reference ligands and acceptors to streptavidin to test this new homogeneous competitive assay. Upon binding of an acceptor to streptavidin, there were the quench of streptavidin fluorescence at 340 nm and the characteristic fluorescence at 430 nm for BNEDA or at 525 nm for BDEDA. Kd of BNEDA and BDEDA was obtained via competitive binding against biotin. By quantifying BNEDA fluorescence, Kd of each tested nonfluorescent biotin derivative was consistent with that by quantifying streptavidin fluorescence using BNEDA or BDEDA as the acceptor. The overall coefficients of variation were about 10%. Therefore, this homogeneous competitive assay was effective and promising to high-throughput-screening.

  10. Determination of metallothioneins by fluorescence and resonance light scattering strategies based on ciprofloxacin–Cu(II) system

    Liu, Lu [College of Public Health, University of South China, Hengyang 421001 (China); Wang, Yong-Sheng, E-mail: yongsheng.w@tom.com [College of Public Health, University of South China, Hengyang 421001 (China); Xue, Jin-Hua; Yang, Hui-Xian; Li, Qiu; Zhou, Bin; Wang, Jia-Cheng; Yin, Ji-Cheng; Wang, Yong-Song [College of Public Health, University of South China, Hengyang 421001 (China); Xiao, Xi-Lin [College of Chemistry and Chemical Engineering, University of South China, Hengyang 421001 (China)

    2013-06-15

    Based on ciprofloxacin (CIP)–Cu(II) system, the novel methods for the detection of metallothioneins (MTs) have been developed by fluorescence (FL) and resonance light scattering (RLS) strategies. The FL strategy avoids the label and derivatization steps in common methods, while the RLS strategy can be applied for determining bio-macromolecules and small molecules without native fluorescence. The response signals linearly correlated with the concentration of MTs over the ranges of 1.03×10{sup −8}–1.23×10{sup −6} mol L{sup −1} for FL, and of 2.56×10{sup −7}–1.54×10{sup −6} mol L{sup −1} for RLS. The limits of detection (LOD) are 3.1×10{sup −9} mol L{sup −1} for FL and 7.68×10{sup −8} mol L{sup −1} for RLS. This study represents the comparison of these two methods using the same CIP–Cu{sup 2+}–MTs system. They not only allow practical application for MTs detection but also serve as a potential choice for the operators according to their concrete needs. In addition, the mechanisms for FL and RLS enhancement of the system were also discussed. -- Highlights: ► Determination of MTs was developed based on CIP–Cu(II) system by FL and RLS strategies. ► FL strategy provides lower limit of detection and wider linear range, and avoids the label and derivatization steps. ► RLS strategy can be applied for determining bio-macromolecules and small molecules. ► The mechanism of interaction of MTs with CIP–Cu(II) chelate was discussed.

  11. Interfacial transduction of nucleic acid hybridization using immobilized quantum dots as donors in fluorescence resonance energy transfer.

    Algar, W Russ; Krull, Ulrich J

    2009-01-06

    Fluorescence resonance energy transfer (FRET) using immobilized quantum dots (QDs) as energy donors was explored as a transduction method for the detection of nucleic acid hybridization at an interface. This research was motivated by the success of the QD-FRET-based transduction of nucleic acid hybridization in solution-phase assays. This new work represents a fundamental step toward the assembly of a biosensor, where immobilization of the selective chemistry on a surface is desired. After immobilizing QD-probe oligonucleotide conjugates on optical fibers, a demonstration of the retention of selectivity was achieved by the introduction of acceptor (Cy3)-labeled single-stranded target oligonucleotides. Hybridization generated the proximity required for FRET, and the resulting fluorescence spectra provided an analytical signal proportional to the amount of target. This research provides an important framework for the future development of nucleic acid biosensors based on QDs and FRET. The most important findings of this work are that (1) a QD-FRET solid-phase hybridization assay is viable and (2) a passivating layer of denatured bovine serum albumin alleviates nonspecific adsorption, ultimately resulting in (3) the potential for a reusable assay format and mismatch discrimination. In this, the first incarnation of a solid-phase QD-FRET hybridization assay, the limit of detection was found to be 5 nM, and the dynamic range was almost 2 orders of magnitude. Selective discrimination of the target was shown using a three-base-pairs mismatch from a fully complementary sequence. Despite a gradual loss of signal, reuse of the optical fibers over multiple cycles of hybridization and dehybridization was possible. Directions for further improvement of the analytical performance by optimizing the design of the QD-probe oligonucleotide interface are identified.

  12. In vivo colocalization of 2-nitroimidazole EF5 fluorescence intensity and electron paramagnetic resonance oximetry in mouse tumors

    Mahy, Pierre; Bast, Marc de; Gallez, Bernard; Gueulette, John; Koch, Cameron J.; Scalliet, Pierre; Gregoire, Vincent

    2003-01-01

    Background and purpose: The primary objective of this study was to establish in vivo the relationship between 2-2-nitro-1H-imidazol-1yl-N-(2,2,3,3,3-pentafluoropropyl)-acetamide (EF5) adduct formation and intratumoral oxygen concentrations measured by electron paramagnetic resonance (EPR) in a tumor model mimicking a clinical situation. The secondary objective was an attempt to calibrate in situ the immunofluorescence (IF) signal with EPR oximetry. Materials and methods: IM syngeneic fibrosarcoma (NFSA) bearing C3H mice were used. Three days after injection of a paramagnetic charcoal into the tumor, the mice were anesthetized, injected with the hypoxic marker EF5, and monitored every 20 min for 3 h with a low-frequency EPR spectrometer. Animals were allowed to breath either under 21 or 100% O 2 . Tumors were then harvested, frozen, cut into sections including the charcoal and processed for EF5 adducts detection using monoclonal antibodies. Slices were viewed with a fluorescence microscope and 190x140 μm areas surrounding the charcoal were digitized and analyzed with the NIH-Image and Adobe Photoshop TM software. The fluorescence intensity (FI) was measured in the whole pictures and in strips of 10 μm around the charcoal. Results: EF5 binding increased with decreasing pO 2 , most substantially at pO 2 below 5 mm Hg. Baseline (ambient air) pO 2 reached 3.2±2.1 mm Hg in NFSA tumors. It increased to 9.8±3.2 mm Hg under 100% O 2 . A statistically significant correlation was observed on an individual tumor basis between the FI in the first 10 μm strip around the charcoal and the pO 2 determined by EPR oximetry (Wilcoxon signed rank test: P 2 in an in vivo environment under biologically-relevant pO 2 values of less than 10 mm Hg

  13. Characterization of the AT180 epitope of phosphorylated Tau protein by a combined nuclear magnetic resonance and fluorescence spectroscopy approach

    Amniai, Laziza; Lippens, Guy; Landrieu, Isabelle

    2011-01-01

    Highlights: → pThr231 of the Tau protein is necessary for the binding of the AT180 antibody. → pSer235 of the Tau protein does not interfere with the AT180 recognition of pThr231. → Epitope mapping is efficiently achieved by combining NMR and FRET spectroscopy. -- Abstract: We present here the characterization of the epitope recognized by the AT180 monoclonal antibody currently used to define an Alzheimer's disease (AD)-related pathological form of the phosphorylated Tau protein. Some ambiguity remains as to the exact phospho-residue(s) recognized by this monoclonal: pThr231 or both pThr231 and pSer235. To answer this question, we have used a combination of nuclear magnetic resonance (NMR) and fluorescence spectroscopy to characterize in a qualitative and quantitative manner the phospho-residue(s) essential for the epitope recognition. Data from the first step of NMR experiments are used to map the residues bound by the antibodies, which were found to be limited to a few residues. A fluorophore is then chemically attached to a cystein residue introduced close-by the mapped epitope, at arginine 221, by mutagenesis of the recombinant protein. The second step of Foerster resonance energy transfer (FRET) between the AT180 antibody tryptophanes and the phospho-Tau protein fluorophore allows to calculate a dissociation constant Kd of 30 nM. We show that the sole pThr231 is necessary for the AT180 recognition of phospho-Tau and that phosphorylation of Ser235 does not interfere with the binding.

  14. Fis protein induced λF-DNA bending observed by single-pair fluorescence resonance energy transfer

    Chi-Cheng, Fu; Wunshain, Fann; Yuan Hanna, S.

    2006-03-01

    Fis, a site-specific DNA binding protein, regulates many biological processes including recombination, transcription, and replication in E.coli. Fis induced DNA bending plays an important role in regulating these functions and bending angle range from ˜50 to 95 dependent on the DNA sequence. For instance, the average bending angle of λF-DNA (26 bp, 8.8nm long, contained λF binding site on the center) measured by gel mobility shift assays was ˜ 94 . But the traditional method cannot provide information about the dynamics and the angle distribution. In this study, λF-DNA was labeled with donor (Alexa Fluor 546) and acceptor (Alexa Fluor 647) dyes on its two 5' ends and the donor-acceptor distances were measured using single-pair fluorescence resonance energy transfer (sp-FRET) with and without the present of Fis protein. Combing with structure information of Fis-DNA complex, the sp-FRET results are used to estimate the protein induced DNA bending angle distribution and dynamics.

  15. Functionalized graphene oxide/Fe3O4 hybrids for cellular magnetic resonance imaging and fluorescence labeling.

    Zhou, Chaohui; Wu, Hui; Wang, Mingliang; Huang, Chusen; Yang, Dapeng; Jia, Nengqin

    2017-09-01

    In this work, we developed a T 2 -weighted contrast agent based on graphene oxide (GO)/Fe 3 O 4 hybrids for efficient cellular magnetic resonance imaging (MRI). The GO/Fe 3 O 4 hybrids were obtained by combining with co-precipitation method and pyrolysis method. The structural, surface and magnetic characteristics of the hybrids were systematically characterized by transmission electron microscopy (TEM), vibrating sample magnetometer (VSM), AFM, Raman, FT-IR and XRD. The GO/Fe 3 O 4 hybrids were functionalized by modifying with anionic and cationic polyelectrolyte through layer-by-layer assembling. The fluorescence probe fluorescein isothiocyanate (FITC) was further loaded on the surface of functionalized GO/Fe 3 O 4 hybrids to trace the location of GO/Fe 3 O 4 hybrids in cells. Functionalized GO/Fe 3 O 4 hybrids possess good hydrophilicity, less cytotoxicity, high MRI enhancement with the relaxivity (r 2 ) of 493mM -1 s -1 as well as cellular MRI contrast effect. These obtained results indicated that the functionalized GO/Fe 3 O 4 hybrids could have great potential to be utilized as cellular MRI contrast agents for tumor early diagnosis and monitoring. Copyright © 2017 Elsevier B.V. All rights reserved.

  16. Nucleic Acid Sandwich Hybridization Assay with Quantum Dot-Induced Fluorescence Resonance Energy Transfer for Pathogen Detection

    Chou, Cheng-Chung; Huang, Yi-Han

    2012-01-01

    This paper reports a nucleic acid sandwich hybridization assay with a quantum dot (QD)-induced fluorescence resonance energy transfer (FRET) reporter system. Two label-free hemagglutinin H5 sequences (60-mer DNA and 630-nt cDNA fragment) of avian influenza viruses were used as the targets in this work. Two oligonucleotides (16 mers and 18 mers) that specifically recognize two separate but neighboring regions of the H5 sequences were served as the capturing and reporter probes, respectively. The capturing probe was conjugated to QD655 (donor) in a molar ratio of 10:1 (probe-to-QD), and the reporter probe was labeled with Alexa Fluor 660 dye (acceptor) during synthesis. The sandwich hybridization assay was done in a 20 μL transparent, adhesive frame-confined microchamber on a disposable, temperature-adjustable indium tin oxide (ITO) glass slide. The FRET signal in response to the sandwich hybridization was monitored by a homemade optical sensor comprising a single 400 nm UV light-emitting diode (LED), optical fibers, and a miniature 16-bit spectrophotometer. The target with a concentration ranging from 0.5 nM to 1 μM was successfully correlated with both QD emission decrease at 653 nm and dye emission increase at 690 nm. To sum up, this work is beneficial for developing a portable QD-based nucleic acid sensor for on-site pathogen detection. PMID:23211753

  17. Development of a Fluorescence Resonance Energy Transfer (FRET)-Based DNA Biosensor for Detection of Synthetic Oligonucleotide of Ganoderma boninense.

    Mohd Bakhori, Noremylia; Yusof, Nor Azah; Abdullah, Abdul Halim; Hussein, Mohd Zobir

    2013-12-01

    An optical DNA biosensor based on fluorescence resonance energy transfer (FRET) utilizing synthesized quantum dot (QD) has been developed for the detection of specific-sequence of DNA for Ganoderma boninense, an oil palm pathogen. Modified QD that contained carboxylic groups was conjugated with a single-stranded DNA probe (ssDNA) via amide-linkage. Hybridization of the target DNA with conjugated QD-ssDNA and reporter probe labeled with Cy5 allows for the detection of related synthetic DNA sequence of Ganoderma boninense gene based on FRET signals. Detection of FRET emission before and after hybridization was confirmed through the capability of the system to produce FRET at 680 nm for hybridized sandwich with complementary target DNA. No FRET emission was observed for non-complementary system. Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system. The developed biosensor has shown high sensitivity with detection limit of 3.55 × 10(-9) M. TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA. This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

  18. A Quantitative Theoretical Framework For Protein-Induced Fluorescence Enhancement-Förster-Type Resonance Energy Transfer (PIFE-FRET).

    Lerner, Eitan; Ploetz, Evelyn; Hohlbein, Johannes; Cordes, Thorben; Weiss, Shimon

    2016-07-07

    Single-molecule, protein-induced fluorescence enhancement (PIFE) serves as a molecular ruler at molecular distances inaccessible to other spectroscopic rulers such as Förster-type resonance energy transfer (FRET) or photoinduced electron transfer. In order to provide two simultaneous measurements of two distances on different molecular length scales for the analysis of macromolecular complexes, we and others recently combined measurements of PIFE and FRET (PIFE-FRET) on the single molecule level. PIFE relies on steric hindrance of the fluorophore Cy3, which is covalently attached to a biomolecule of interest, to rotate out of an excited-state trans isomer to the cis isomer through a 90° intermediate. In this work, we provide a theoretical framework that accounts for relevant photophysical and kinetic parameters of PIFE-FRET, show how this framework allows the extraction of the fold-decrease in isomerization mobility from experimental data, and show how these results provide information on changes in the accessible volume of Cy3. The utility of this model is then demonstrated for experimental results on PIFE-FRET measurement of different protein-DNA interactions. The proposed model and extracted parameters could serve as a benchmark to allow quantitative comparison of PIFE effects in different biological systems.

  19. Development of a Fluorescence Resonance Energy Transfer (FRET-Based DNA Biosensor for Detection of Synthetic Oligonucleotide of Ganoderma boninense

    Noremylia Mohd Bakhori

    2013-12-01

    Full Text Available An optical DNA biosensor based on fluorescence resonance energy transfer (FRET utilizing synthesized quantum dot (QD has been developed for the detection of specific-sequence of DNA for Ganoderma boninense, an oil palm pathogen. Modified QD that contained carboxylic groups was conjugated with a single-stranded DNA probe (ssDNA via amide-linkage. Hybridization of the target DNA with conjugated QD-ssDNA and reporter probe labeled with Cy5 allows for the detection of related synthetic DNA sequence of Ganoderma boninense gene based on FRET signals. Detection of FRET emission before and after hybridization was confirmed through the capability of the system to produce FRET at 680 nm for hybridized sandwich with complementary target DNA. No FRET emission was observed for non-complementary system. Hybridization time, temperature and effect of different concentration of target DNA were studied in order to optimize the developed system. The developed biosensor has shown high sensitivity with detection limit of 3.55 × 10−9 M. TEM results show that the particle size of QD varies in the range between 5 to 8 nm after ligand modification and conjugation with ssDNA. This approach is capable of providing a simple, rapid and sensitive method for detection of related synthetic DNA sequence of Ganoderma boninense.

  20. Resonance

    Petersen, Nils Holger

    2014-01-01

    A chapter in a book about terminology within the field of medievalism: the chapter discusses the resonance of medieval music and ritual in modern (classical) music culture and liturgical practice.......A chapter in a book about terminology within the field of medievalism: the chapter discusses the resonance of medieval music and ritual in modern (classical) music culture and liturgical practice....

  1. A ratiometric fluorescent probe based on boron dipyrromethene and rhodamine Förster resonance energy transfer platform for hypochlorous acid and its application in living cells

    Liu, Ying; Zhao, Zhi-Min; Miao, Jun-Ying; Zhao, Bao-Xiang

    2016-01-01

    We have developed a ratiometric fluorescent probe BRT based on boron dipyrromethene (BODIPY) and rhodamine-thiohydrazide Förster resonance energy transfer (FRET) platform for sensing hypochlorous acid (HOCl) with high selectivity and sensitivity. The probe can detect HOCl in 15 s with the detection limit of 38 nM. Upon mixing with HOCl the fluorescence colour of probe BRT changed from green to orange. Moreover, probe BRT was applied to successfully monitor HOCl in living RAW 264.7 cells. - Highlights: • A probe based on BODIPY and rhodamine was developed for sensing HOCl. • The probe could sense HOCl in a ratiometric manner based on the FRET platform in PBS buffer solution. • The probe can detect HOCl in 15 s accompanied with a fluorescence colour change. • This probe was successfully used to monitor HOCl in living RAW 264.7 cells.

  2. Phosphorylation-induced conformational changes in short peptides probed by fluorescence resonance energy transfer in the 10A domain.

    Sahoo, Harekrushna; Nau, Werner M

    2007-03-26

    Phosphorylation-induced conformational changes in short polypeptides were probed by a fluorescence resonance energy transfer (FRET) method by employing a short-distance FRET pair (R(0) approximately 10 A) based on tryptophan as natural donor and a 2,3-diazabicyclo[2.2.2]oct-2-ene-labeled asparagine (Dbo) as synthetic acceptor. Two substrates for kinases, LeuArgArgTrpSerLeuGly-Dbo (peptide I) and TrpLysArgThrLeuArgArg-Dbo (peptide II), were investigated, with serine and threonine, respectively, as phosphorylation sites. Steady-state and time-resolved fluorescence experiments in H(2)O revealed a decrease in FRET efficiency for peptide I and an increase for peptide II; this suggested that the effective distances between donor and acceptor increased and decreased, respectively. The same trends and similar absolute variations in effective donor-acceptor distances were observed in propylene glycol, a less polar and highly viscous solvent; this suggested that the variations are due to intrinsic structural preferences. Fitting of the time-resolved decay traces according to a distribution function model (Gaussian distribution) provided the mean donor-acceptor distances, which showed an increase upon phosphorylation for peptide I (from 9.7 to 10.5 A) and a decrease for peptide II (from 10.9 to 9.3 A) in H(2)O. The broadness (half-width) of the distributions, which provides a measure of the rigidity of the peptides, remained similar upon phosphorylation of peptide I (3.0 versus 3.1 A), but decreased for peptide II (from 3.1 to 0.73 A in H(2)O); this suggests a more compact, structured conformation upon phosphorylation of the latter peptide. The elongation of the peptide backbone (by ca. 0.7 A) for peptide I is attributed to an increase in steric demand upon phosphorylation, which favors an extended conformation. The contraction (by ca. 1.4 A) and structural rigidification of peptide II is attributed to attractive Coulombic interactions and hydrogen bonding between the

  3. Determination of trace uranium by resonance fluorescence method coupled with photo-catalytic technology and dual cloud point extraction.

    Li, Jiekang; Li, Guirong; Han, Qian

    2016-12-05

    In this paper, two kinds of salophens (Sal) with different solubilities, Sal1 and Sal2, have been respectively synthesized, and they all can combine with uranyl to form stable complexes: [UO2(2+)-Sal1] and [UO2(2+)-Sal2]. Among them, [UO2(2+)-Sal1] was used as ligand to extract uranium in complex samples by dual cloud point extraction (dCPE), and [UO2(2+)-Sal2] was used as catalyst for the determination of uranium by photocatalytic resonance fluorescence (RF) method. The photocatalytic characteristic of [UO2(2+)-Sal2] on the oxidized pyronine Y (PRY) by potassium bromate which leads to the decrease of RF intensity of PRY were studied. The reduced value of RF intensity of reaction system (ΔF) is in proportional to the concentration of uranium (c), and a novel photo-catalytic RF method was developed for the determination of trace uranium (VI) after dCPE. The combination of photo-catalytic RF techniques and dCPE procedure endows the presented methods with enhanced sensitivity and selectivity. Under optimal conditions, the linear calibration curves range for 0.067 to 6.57ngmL(-1), the linear regression equation was ΔF=438.0 c (ngmL(-1))+175.6 with the correlation coefficient r=0.9981. The limit of detection was 0.066ngmL(-1). The proposed method was successfully applied for the separation and determination of uranium in real samples with the recoveries of 95.0-103.5%. The mechanisms of the indicator reaction and dCPE are discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

  4. Effect of enhanced Renilla luciferase and fluorescent protein variants on the Foerster distance of Bioluminescence resonance energy transfer (BRET)

    Dacres, Helen, E-mail: helen.dacres@csiro.au [CSIRO Food Futures Flagship and Ecosystem Sciences, Canberra (Australia); Michie, Michelle; Wang, Jian [CSIRO Food Futures Flagship and Ecosystem Sciences, Canberra (Australia); Pfleger, Kevin D.G. [Laboratory for Molecular Endocrinology-GPCRs, Western Australian Institute for Medical Research (WAIMR) and Centre for Medical Research, The University of Western Australia, Perth (Australia); Trowell, Stephen C. [CSIRO Food Futures Flagship and Ecosystem Sciences, Canberra (Australia)

    2012-08-31

    Highlights: Black-Right-Pointing-Pointer First experimental determination of Foerster distance (R{sub 0}) for enhanced BRET systems. Black-Right-Pointing-Pointer Effect of brighter BRET components RLuc2, RLuc8 and Venus was assessed. Black-Right-Pointing-Pointer Using brighter BRET components substantially increased (25%) R{sub 0} of the BRET{sup 1} system. Black-Right-Pointing-Pointer Using brighter BRET components marginally increased (2-9%) R{sub 0} of the BRET{sup 2} system. Black-Right-Pointing-Pointer Brighter BRET components improve the different weaknesses of BRET{sup 1} and BRET{sup 2} systems. -- Abstract: Bioluminescence resonance energy transfer (BRET) is an important tool for monitoring macromolecular interactions and is useful as a transduction technique for biosensor development. Foerster distance (R{sub 0}), the intermolecular separation characterized by 50% of the maximum possible energy transfer, is a critical BRET parameter. R{sub 0} provides a means of linking measured changes in BRET ratio to a physical dimension scale and allows estimation of the range of distances that can be measured by any donor-acceptor pair. The sensitivity of BRET assays has recently been improved by introduction of new BRET components, RLuc2, RLuc8 and Venus with improved quantum yields, stability and brightness. We determined R{sub 0} for BRET{sup 1} systems incorporating novel RLuc variants RLuc2 or RLuc8, in combination with Venus, as 5.68 or 5.55 nm respectively. These values were approximately 25% higher than the R{sub 0} of the original BRET{sup 1} system. R{sub 0} for BRET{sup 2} systems combining green fluorescent proteins (GFP{sup 2}) with RLuc2 or RLuc8 variants was 7.67 or 8.15 nm, i.e. only 2-9% greater than the original BRET{sup 2} system despite being {approx}30-fold brighter.

  5. Application of the laser induced fluorescence to the investigation of highly magnetized plasmas, heated by ion cyclotron resonance

    Pailloux, A.

    1997-01-01

    This work has been achieved in the frame of isotopic separation studies by in cyclotron resonance. For this purpose, in a highly magnetized (2 to 3 Tesla) and non-collisional (10 12 ions/cm 3 ) plasma, composed of metallic ions, a wave near the ion cyclotron frequency is thrown in order to heat selectively a given species. A laser induced fluorescence (LIP) has been developed on barium and gadolinium plasmas. The Larmor gyration of ions greatly modifies the interaction, which has been modelled through the time-dependent Schroedinger equation. The obtained excitation probably has been integrated over all the ions excited in the measurement volume in order to check that the LIF still leads to the distribution function of ion velocities. The influence of the Larmor motion of ions on the spectral distribution of LIF has been derived both theoretically and experimentally. The LIF diagnostics has been achieved with a dye O'ring laser. The barium ion has been excited on the transition 6142 angstrom, using rhodamine 6G dye, and the gadolinium ion on the pseudo-triplet 3861 angstrom, using exalite dye. Data treatment has been developed taking into account the Zeeman effect and the different heating of isotopes. The ionic temperature (from 1 eV to some hundreds eV) has been measured as a function of radiofrequency heating. Our experimental results are in good agreement with the selective heating theory. Also, the ion velocity distribution function has been found locally Maxwellian. And the behaviour of the plasma has been studied as a function of control parameters of the plasma source. (author)

  6. Near-infrared Fluorescence Optical Imaging in Early Rheumatoid Arthritis: A Comparison to Magnetic Resonance Imaging and Ultrasonography.

    Krohn, Michaela; Ohrndorf, Sarah; Werner, Stephanie G; Schicke, Bernd; Burmester, Gerd-Rüdiger; Hamm, Bernd; Backhaus, Marina; Hermann, Kay-Geert A

    2015-07-01

    Near-infrared fluorescence optical imaging (FOI) is a novel imaging technology in the detection and evaluation of different arthritides. FOI was validated in comparison to magnetic resonance imaging (MRI), greyscale ultrasonography (GSUS), and power Doppler ultrasonography (PDUS) in patients with early rheumatoid arthritis (RA). Hands of 31 patients with early RA were examined by FOI, MRI, and US. In each modality, synovitis of the wrist, metacarpophalangeal joints (MCP) 2-5, and proximal interphalangeal joints (PIP) 2-5 were scored on a 4-point scale (0-3). Sensitivity and specificity of FOI were analyzed in comparison to MRI and US as reference methods, differentiating between 3 phases of FOI enhancement (P1-3). Intraclass correlation coefficients (ICC) were calculated to evaluate the agreement of FOI with MRI and US. A total of 279 joints (31 wrists, 124 MCP and 124 PIP joints) were evaluated. With MRI as the reference method, overall sensitivity/specificity of FOI was 0.81/0.00, 0.49/0.84, and 0.86/0.38 for wrist, MCP, and PIP joints, respectively. Under application of PDUS as reference, sensitivity was even higher, while specificity turned out to be low, except for MCP joints (0.88/0.15, 0.81/0.76, and 1.00/0.27, respectively). P2 appears to be the most sensitive FOI phase, while P1 showed the highest specificity. The best agreement of FOI was shown for PDUS, especially with regard to MCP and PIP joints (ICC of 0.57 and 0.53, respectively), while correlation with MRI was slightly lower. FOI remains an interesting diagnostic tool for patients with early RA, although this study revealed limitations concerning the detection of synovitis. Further research is needed to evaluate its full diagnostic potential in rheumatic diseases.

  7. A high-throughput fluorescence resonance energy transfer (FRET)-based endothelial cell apoptosis assay and its application for screening vascular disrupting agents

    Zhu, Xiaoming; Fu, Afu; Luo, Kathy Qian

    2012-01-01

    Highlights: ► An endothelial cell apoptosis assay using FRET-based biosensor was developed. ► The fluorescence of the cells changed from green to blue during apoptosis. ► This method was developed into a high-throughput assay in 96-well plates. ► This assay was applied to screen vascular disrupting agents. -- Abstract: In this study, we developed a high-throughput endothelial cell apoptosis assay using a fluorescence resonance energy transfer (FRET)-based biosensor. After exposure to apoptotic inducer UV-irradiation or anticancer drugs such as paclitaxel, the fluorescence of the cells changed from green to blue. We developed this method into a high-throughput assay in 96-well plates by measuring the emission ratio of yellow fluorescent protein (YFP) to cyan fluorescent protein (CFP) to monitor the activation of a key protease, caspase-3, during apoptosis. The Z′ factor for this assay was above 0.5 which indicates that this assay is suitable for a high-throughput analysis. Finally, we applied this functional high-throughput assay for screening vascular disrupting agents (VDA) which could induce endothelial cell apoptosis from our in-house compounds library and dioscin was identified as a hit. As this assay allows real time and sensitive detection of cell apoptosis, it will be a useful tool for monitoring endothelial cell apoptosis in living cell situation and for identifying new VDA candidates via a high-throughput screening.

  8. In vivo magnetic resonance and fluorescence dual imaging of tumor sites by using dye-doped silica-coated iron oxide nanoparticles

    Jang, Haeyun; Lee, Chaedong; Nam, Gi-Eun; Quan, Bo; Choi, Hyuck Jae; Yoo, Jung Sun; Piao, Yuanzhe

    2016-01-01

    The difficulty in delineating tumor is a major obstacle for better outcomes in cancer treatment of patients. The use of single-imaging modality is often limited by inadequate sensitivity and resolution. Here, we present the synthesis and the use of monodisperse iron oxide nanoparticles coated with fluorescent silica nano-shells for fluorescence and magnetic resonance dual imaging of tumor. The as-synthesized core–shell nanoparticles were designed to improve the accuracy of diagnosis via simultaneous tumor imaging with dual imaging modalities by a single injection of contrast agent. The iron oxide nanocrystals (∼11 nm) were coated with Rhodamine B isothiocyanate-doped silica shells via reverse microemulsion method. Then, the core–shell nanoparticles (∼54 nm) were analyzed to confirm their size distribution by transmission electron microscopy and dynamic laser scattering. Photoluminescence spectroscopy was used to characterize the fluorescent property of the dye-doped silica shell-coated nanoparticles. The cellular compatibility of the as-prepared nanoparticles was confirmed by a trypan blue dye exclusion assay and the potential as a dual-imaging contrast agent was verified by in vivo fluorescence and magnetic resonance imaging. The experimental results show that the uniform-sized core–shell nanoparticles are highly water dispersible and the cellular toxicity of the nanoparticles is negligible. In vivo fluorescence imaging demonstrates the capability of the developed nanoparticles to selectively target tumors by the enhanced permeability and retention effects and ex vivo tissue analysis was corroborated this. Through in vitro phantom test, the core/shell nanoparticles showed a T2 relaxation time comparable to Feridex ® with smaller size, indicating that the as-made nanoparticles are suitable for imaging tumor. This new dual-modality-nanoparticle approach has promised for enabling more accurate tumor imaging.

  9. In vivo magnetic resonance and fluorescence dual imaging of tumor sites by using dye-doped silica-coated iron oxide nanoparticles

    Jang, Haeyun; Lee, Chaedong [Seoul National University, Program in Nano Science and Technology, Graduate School of Convergence Science and Technology (Korea, Republic of); Nam, Gi-Eun [University of Ulsan College of Medicine, Department of Radiology, Asan Medical Center (Korea, Republic of); Quan, Bo [Seoul National University, Program in Nano Science and Technology, Graduate School of Convergence Science and Technology (Korea, Republic of); Choi, Hyuck Jae [University of Ulsan College of Medicine, Department of Radiology, Asan Medical Center (Korea, Republic of); Yoo, Jung Sun [Seoul National University, Department of Transdisciplinary Studies, Graduate School of Convergence Science and Technology, Smart Humanity Convergence Center (Korea, Republic of); Piao, Yuanzhe, E-mail: parkat9@snu.ac.kr [Seoul National University, Program in Nano Science and Technology, Graduate School of Convergence Science and Technology (Korea, Republic of)

    2016-02-15

    The difficulty in delineating tumor is a major obstacle for better outcomes in cancer treatment of patients. The use of single-imaging modality is often limited by inadequate sensitivity and resolution. Here, we present the synthesis and the use of monodisperse iron oxide nanoparticles coated with fluorescent silica nano-shells for fluorescence and magnetic resonance dual imaging of tumor. The as-synthesized core–shell nanoparticles were designed to improve the accuracy of diagnosis via simultaneous tumor imaging with dual imaging modalities by a single injection of contrast agent. The iron oxide nanocrystals (∼11 nm) were coated with Rhodamine B isothiocyanate-doped silica shells via reverse microemulsion method. Then, the core–shell nanoparticles (∼54 nm) were analyzed to confirm their size distribution by transmission electron microscopy and dynamic laser scattering. Photoluminescence spectroscopy was used to characterize the fluorescent property of the dye-doped silica shell-coated nanoparticles. The cellular compatibility of the as-prepared nanoparticles was confirmed by a trypan blue dye exclusion assay and the potential as a dual-imaging contrast agent was verified by in vivo fluorescence and magnetic resonance imaging. The experimental results show that the uniform-sized core–shell nanoparticles are highly water dispersible and the cellular toxicity of the nanoparticles is negligible. In vivo fluorescence imaging demonstrates the capability of the developed nanoparticles to selectively target tumors by the enhanced permeability and retention effects and ex vivo tissue analysis was corroborated this. Through in vitro phantom test, the core/shell nanoparticles showed a T2 relaxation time comparable to Feridex{sup ®} with smaller size, indicating that the as-made nanoparticles are suitable for imaging tumor. This new dual-modality-nanoparticle approach has promised for enabling more accurate tumor imaging.

  10. Resonances

    an impetus or drive to that account: change, innovation, rupture, or discontinuity. Resonances: Historical Essays on Continuity and Change explores the historiographical question of the modes of interrelation between these motifs in historical narratives. The essays in the collection attempt to realize...

  11. A highly sensitive fluorescence resonance energy transfer aptasensor for staphylococcal enterotoxin B detection based on exonuclease-catalyzed target recycling strategy

    Wu, Shijia; Duan, Nuo; Ma, Xiaoyuan; Xia, Yu; Wang, Hongxin; Wang, Zhouping

    2013-01-01

    Graphical abstract: -- Highlights: •An ultrasensitive FRET aptasensor was developed for staphylococcal enterotoxin B determination. •SEB was recognized by SEB aptamer with high affinity and specificity. •The Mn 2+ doped NaYF 4 :Yb/Er UCNPs used as donor to quencher dye (BHQ 3 ) in new FRET. •The fluorescence intensity was prominently amplified using an exonuclease-catalyzed target recycling strategy. -- Abstract: An ultrasensitive fluorescence resonance energy transfer (FRET) bioassay was developed to detect staphylococcal enterotoxin B (SEB), a low molecular exotoxin, using an aptamer-affinity method coupled with upconversion nanoparticles (UCNPs)-sensing, and the fluorescence intensity was prominently enhanced using an exonuclease-catalyzed target recycling strategy. To construct this aptasensor, both fluorescence donor probes (complementary DNA 1 –UCNPs) and fluorescence quencher probes (complementary DNA 2 –Black Hole Quencher 3 (BHQ 3 )) were hybridized to an SEB aptamer, and double-strand oligonucleotides were fabricated, which quenched the fluorescence of the UCNPs via FRET. The formation of an aptamer–SEB complex in the presence of the SEB analyte resulted in not only the dissociation of aptamer from the double-strand DNA but also both the disruption of the FRET system and the restoration of the UCNPs fluorescence. In addition, the SEB was liberated from the aptamer–SEB complex using exonuclease I, an exonuclease specific to single-stranded DNA, for analyte recycling by selectively digesting a particular DNA (SEB aptamer). Based on this exonuclease-catalyzed target recycling strategy, an amplified fluorescence intensity could be produced using different SEB concentrations. Using optimized experimental conditions produced an ultrasensitive aptasensor for the detection of SEB, with a wide linear range of 0.001–1 ng mL −1 and a lower detection limit (LOD) of 0.3 pg mL −1 SEB (at 3σ). The fabricated aptasensor was used to measure SEB in a

  12. Double-labeled donor probe can enhance the signal of fluorescence resonance energy transfer (FRET) in detection of nucleic acid hybridization

    Okamura, Yukio; Kondo, Satoshi; Sase, Ichiro; Suga, Takayuki; Mise, Kazuyuki; Furusawa, Iwao; Kawakami, Shigeki; Watanabe, Yuichiro

    2000-01-01

    A set of fluorescently-labeled DNA probes that hybridize with the target RNA and produce fluorescence resonance energy transfer (FRET) signals can be utilized for the detection of specific RNA. We have developed probe sets to detect and discriminate single-strand RNA molecules of plant viral genome, and sought a method to improve the FRET signals to handle in vivo applications. Consequently, we found that a double-labeled donor probe labeled with Bodipy dye yielded a remarkable increase in fluorescence intensity compared to a single-labeled donor probe used in an ordinary FRET. This double-labeled donor system can be easily applied to improve various FRET probes since the dependence upon sequence and label position in enhancement is not as strict. Furthermore this method could be applied to other nucleic acid substances, such as oligo RNA and phosphorothioate oligonucleotides (S-oligos) to enhance FRET signal. Although the double-labeled donor probes labeled with a variety of fluorophores had unexpected properties (strange UV-visible absorption spectra, decrease of intensity and decay of donor fluorescence) compared with single-labeled ones, they had no relation to FRET enhancement. This signal amplification mechanism cannot be explained simply based on our current results and knowledge of FRET. Yet it is possible to utilize this double-labeled donor system in various applications of FRET as a simple signal-enhancement method. PMID:11121494

  13. Self-Assembled Fluorescent Nanoprobe Based on Forster Resonance Energy Transfer for Carbon Monoxide in Living Cells and Animals via Ligand Exchange.

    Jia, Ruizhen; Song, Pengfei; Wang, Jingjing; Mai, Hengtang; Li, Sixian; Cheng, Yu; Wu, Song

    2018-05-29

    Carbon monoxide (CO) is recognized as a biologically essential gaseous neurotransmitter that modulates many physiological processes in living subjects. Currently reported fluorescent probes for CO imaging in cells basically utilize palladium related chemistry which requires complicated synthetic work. Herein we provide a new strategy to construct a fluorescent nanoprobe, NanoCO-1, based on the Forster resonance energy transfer (FRET) mechanism by entrapping the existing dirhodium complex as the energy acceptor and the CO recognition part, and a commonly used nitrobenzoxadiazole (NBD) dye as energy donor into a micelle formed by self-assembly. The exchange of ligands in the dirhodium complex by CO in the nanoprobe disrupts the FRET and leads to the turn-on of fluorescence. The merits of NanoCO-1 including good biocompatibility, selectivity, photostability, and low cytotoxity, render this nanoprobe ability to track CO in living cells, zebrafish embryo, and larvae. Our straightforward approach can be extended to establish the CO fluorescent probes based on adsorption of CO on a variety of metal derivatives.

  14. Gadolinium- and manganite-based contrast agents with fluorescent probes for both magnetic resonance and fluorescence imaging of pancreatic islets: a comparative study

    Berková, Z.; Jirák, D.; Zacharovová, K.; Lukeš, I.; Kotková, Z.; Kotek, J.; Kačenka, M.; Kaman, Ondřej; Řehoř, I.; Hájek, M.; Saudek, F.

    2013-01-01

    Roč. 8, č. 4 (2013), s. 614-621 ISSN 1860-7179 Institutional support: RVO:68378271 Keywords : contrast agents * gadolinium * magnetic resonance imaging * manganite * pancreatic islet s Subject RIV: BM - Solid Matter Physics ; Magnetism Impact factor: 3.046, year: 2013

  15. Targeting human c-Myc promoter duplex DNA with actinomycin D by use of multi-way analysis of quantum-dot-mediated fluorescence resonance energy transfer

    Gholami, Somayeh; Kompany Zare, Mohsen

    2013-01-01

    Actinomycin D (Act D), an oncogenic c-Myc promoter binder, interferes with the action of RNA polymerase. There is great demand for high-throughput technology able to monitor the activity of DNA-binding drugs. To this end, binding of 7-aminoactinomycin D (7AAD) to the duplex c-Myc promoter...... pairs resulted in efficient energy transfer from drug to QD via fluorescence resonance energy transfer (FRET). Multi-way analysis of the three-way data array obtained from titration experiments was performed by use of restricted Tucker3 and hard trilinear decomposition (HTD). These techniques enable...... the important advantage over univariate classical methods of enabling us to investigate the source of variance in the fluorescence signal of the DNA-drug complex. It was established that hard trilinear decomposition analysis of FRET-measured data overcomes the problem of rank deficiency, enabling calculation...

  16. Fluorescence resonance energy transfer between perylene and riboflavin in micellar solution and analytical application on determination of vitamin B{sub 2}

    Bhattar, S.L.; Kolekar, G.B. [Fluorescence Spectroscopy Research Laboratory, Department of Chemistry, Shivaji University, Kolhapur 416 004, Maharashtra (India); Patil, S.R. [Fluorescence Spectroscopy Research Laboratory, Department of Chemistry, Shivaji University, Kolhapur 416 004, Maharashtra (India)], E-mail: srp_fsl@rediffmail.com

    2008-03-15

    Fluorescence resonance energy transfer (FRET) between perylene and riboflavin is studied in micellar solution of sodium dodecyl sulfate. The fluorescence of perylene is quenched by riboflavin and quenching is in accordance with Stern-Volmer relation. The efficiency of energy transfer is found to depend on the concentration of riboflavin. The value of critical energy transfer distance (R{sub 0}) calculated by using Foster relation is 32.13 A, and as it is less than 50 A, it indicates efficient energy transfer in the present system. The analytical relation was established between extent of sensitization and concentration of riboflavin, which helped to estimate vitamin B{sub 2} directly from pharmaceutical tablets.

  17. Quantitative time domain analysis of lifetime-based Förster resonant energy transfer measurements with fluorescent proteins: Static random isotropic fluorophore orientation distributions

    Alexandrov, Yuriy; Nikolic, Dino Solar; Dunsby, Christopher

    2018-01-01

    Förster resonant energy transfer (FRET) measurements are widely used to obtain information about molecular interactions and conformations through the dependence of FRET efficiency on the proximity of donor and acceptor fluorophores. Fluorescence lifetime measurements can provide quantitative...... into new software for fitting donor emission decay profiles. Calculated FRET parameters, including molar population fractions, are compared for the analysis of simulated and experimental FRET data under the assumption of static and dynamic fluorophores and the intermediate regimes between fully dynamic...... analysis of FRET efficiency and interacting population fraction. Many FRET experiments exploit the highly specific labelling of genetically expressed fluorescent proteins, applicable in live cells and organisms. Unfortunately, the typical assumption of fast randomization of fluorophore orientations...

  18. The convergence of quantum-dot-mediated fluorescence resonance energy transfer and microfluidics for monitoring DNA polyplex self-assembly in real time

    Ho Yiping; Wang, T-H; Chen, Hunter H; Leong, Kam W

    2009-01-01

    We present a novel convergence of quantum-dot-mediated fluorescence resonance energy transfer (QD-FRET) and microfluidics, through which molecular interactions were precisely controlled and monitored using highly sensitive quantum-dot-mediated FRET. We demonstrate its potential in studying the kinetics of self-assembly of DNA polyplexes under laminar flow in real time with millisecond resolution. The integration of nanophotonics and microfluidics offers a powerful tool for elucidating the formation of polyelectrolyte polyplexes, which is expected to provide better control and synthesis of uniform and customizable polyplexes for future nucleic acid-based therapeutics.

  19. CH3 NH3 PbBr3 Perovskite Nanocrystals as Efficient Light-Harvesting Antenna for Fluorescence Resonance Energy Transfer.

    Muthu, Chinnadurai; Vijayan, Anuja; Nair, Vijayakumar C

    2017-05-04

    Hybrid perovskites have created enormous research interest as a low-cost material for high-performance photovoltaic devices, light-emitting diodes, photodetectors, memory devices and sensors. Perovskite materials in nanocrystal form that display intense luminescence due to the quantum confinement effect were found to be particularly suitable for most of these applications. However, the potential use of perovskite nanocrystals as a light-harvesting antenna for possible applications in artificial photosynthesis systems is not yet explored. In the present work, we study the light-harvesting antenna properties of luminescent methylammonium lead bromide (CH 3 NH 3 PbBr 3 )-based perovskite nanocrystals using fluorescent dyes (rhodamine B, rhodamine 101, and nile red) as energy acceptors. Our studies revealed that CH 3 NH 3 PbBr 3 nanocrystals are an excellent light-harvesting antenna, and efficient fluorescence resonance energy transfer occurs from the nanocrystals to fluorescent dyes. Further, the energy transfer efficiency is found to be highly dependent on the number of anchoring groups and binding ability of the dyes to the surface of the nanocrystals. These observations may have significant implications for perovskite-based light-harvesting devices and their possible use in artificial photosynthesis systems. © 2017 Wiley-VCH Verlag GmbH & Co. KGaA, Weinheim.

  20. 1,3-Bis(2-chloroethyl)-1-nitrosourea-loaded bovine serum albumin nanoparticles with dual magnetic resonance-fluorescence imaging for tracking of chemotherapeutic agents.

    Wei, Kuo-Chen; Lin, Feng-Wei; Huang, Chiung-Yin; Ma, Chen-Chi M; Chen, Ju-Yu; Feng, Li-Ying; Yang, Hung-Wei

    To date, knowing how to identify the location of chemotherapeutic agents in the human body after injection is still a challenge. Therefore, it is urgent to develop a drug delivery system with molecular imaging tracking ability to accurately understand the distribution, location, and concentration of a drug in living organisms. In this study, we developed bovine serum albumin (BSA)-based nanoparticles (NPs) with dual magnetic resonance (MR) and fluorescence imaging modalities (fluorescein isothiocyanate [FITC]-BSA-Gd/1,3-bis(2-chloroethyl)-1-nitrosourea [BCNU] NPs) to deliver BCNU for inhibition of brain tumor cells (MBR 261-2). These BSA-based NPs are water dispersible, stable, and biocompatible as confirmed by XTT cell viability assay. In vitro phantoms and in vivo MR and fluorescence imaging experiments show that the developed FITC-BSA-Gd/BCNU NPs enable dual MR and fluorescence imaging for monitoring cellular uptake and distribution in tumors. The T1 relaxivity (R1) of FITC-BSA-Gd/BCNU NPs was 3.25 mM(-1) s(-1), which was similar to that of the commercial T1 contrast agent (R1 =3.36 mM(-1) s(-1)). The results indicate that this multifunctional drug delivery system has potential bioimaging tracking of chemotherapeutic agents ability in vitro and in vivo for cancer therapy.

  1. A graphene oxide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of botulinum neurotoxin A (BoNT/A) enzymatic activity.

    Shi, Jingyu; Guo, Jiubiao; Bai, Gongxun; Chan, Chunyu; Liu, Xuan; Ye, Weiwei; Hao, Jianhua; Chen, Sheng; Yang, Mo

    2015-03-15

    Botulinum neurotoxins (BoNTs) are among the most potent toxic bacterial proteins for humans, which make them potential agents for bioterrorism. Therefore, an ultrasensitive detection of BoNTs and their active states is in great need as field-deployable systems for anti-terrorism applications. We report the construction of a novel graphene oxide (GO)-peptide based fluorescence resonance energy transfer (FRET) biosensor for ultrasensitive detection of the BoNT serotype A light chain (BoNT-LcA) protease activity. A green fluorescence protein (GFP) modified SNAP-25 peptide substrate (SNAP-25-GFP) was optimally designed and synthesized with the centralized recognition/cleavage sites. This FRET platform was constructed by covalent immobilization of peptide substrate on GO with BSA passivation which have advantages of low non-specific adsorption and high stability in protein abundant solution. BoNT-LcA can specifically cleave SNAP-25-GFP substrate covalently immobilized on GO to release the fragment with GFP. Based on fluorescence signal recovery measurement, the target BoNT-LcA was detected sensitively and selectively with the linear detection range from 1fg/mL to 1pg/mL. The limit of detection (LOD) for BoNT-LcA is around 1fg/mL. Copyright © 2014 Elsevier B.V. All rights reserved.

  2. Role of scattering processes in spectrum formation of multi-quantum resonant fluorescence of a hydrogen-like system

    Prepelitsa, O.B.

    1996-01-01

    The two-level system with degenerated excitation state, interacting with a coherent electromagnetic field, is considered. It is shown that the fluorescence spectrum consists of the multitude of Mollow triplets. The intensities of components of each triplet are the nonlinear functions of the electromagnetic field intensity. 11 refs

  3. Selective fluorescence resonance energy transfer from serum albumins to a bio-active 3-pyrazolyl-2-pyrazoline derivative: A spectroscopic analysis

    Sarkar, Arindam [Department of Chemistry, Jadavpur University, Kolkata 700032 (India); Bhattacharya, Subhash Chandra, E-mail: sbjuchem@yahoo.com [Department of Chemistry, Jadavpur University, Kolkata 700032 (India)

    2012-10-15

    A novel fluorescent probe and pharmaceutically significant: 3-pyrazolyl-2-pyrazoline derivative (PYZ) has been selected as an acceptor molecule for fluorescence resonance energy transfer (FRET) interaction with serum albumins. Steady state and time resolved fluorescence techniques were applied to elucidate the nature of interaction of PYZ with serum albumins (BSA and HSA). Negligible FRET mediated emission occurred in the case of HSA but an efficient FRET mediated emission resulted in case of BSA. To gain further insight into the FRET selectivity of PYZ with the proteins, FRET from L-tryptophan (donor; native tryptophan) to PYZ (acceptor) was performed with the aim of getting an idea about the steric restrictions imposed on PYZ by the other groups present in BSA and HSA. The studies revealed that the surface bound Trp-134 in BSA allows an efficient FRET process with PYZ while the buried Trp-214 in HSA does not. The unusual selectivity for FRET in case of PYZ and the serum albumins has also been attributed to the complex structure of PYZ due to the presence of bulkier phenyl moieties in it. The complex nature of the excited state photophysics of tryptophan (Trp) in proteins also accounts for this FRET selectivity of PYZ with BSA and HSA. - Highlights: Black-Right-Pointing-Pointer FRET from BSA/HSA to PYZ was monitored using steady state and time resolved fluorescence methods. Black-Right-Pointing-Pointer Efficient FRET process resulted in BSA-PYZ system in contrast with the HSA-PYZ system. Black-Right-Pointing-Pointer Surface bound Trp-134 in BSA facilitates the FRET process with PYZ than the buried Trp-214 in HSA. Black-Right-Pointing-Pointer Rigid and complex structure of PYZ also accounts for the FRET selectivity of PYZ with BSA/HSA.

  4. Three-color confocal Förster (or fluorescence) resonance energy transfer microscopy: Quantitative analysis of protein interactions in the nucleation of actin filaments in live cells.

    Wallrabe, Horst; Sun, Yuansheng; Fang, Xiaolan; Periasamy, Ammasi; Bloom, George S

    2015-06-01

    Experiments using live cell 3-color Förster (or fluorescence) resonance energy transfer (FRET) microscopy and corresponding in vitro biochemical reconstitution of the same proteins were conducted to evaluate actin filament nucleation. A novel application of 3-color FRET data is demonstrated, extending the analysis beyond the customary energy-transfer efficiency (E%) calculations. MDCK cells were transfected for coexpression of Teal-N-WASP/Venus-IQGAP1/mRFP1-Rac1, Teal-N-WASP/Venus-IQGAP1/mRFP1-Cdc42, CFP-Rac1/Venus-IQGAP1/mCherry-actin, or CFP-Cdc42/Venus-IQGAP1/mCherry-actin, and with single-label equivalents for spectral bleedthrough correction. Using confirmed E% as an entry point, fluorescence levels and related ratios were correlated at discrete accumulating levels at cell peripheries. Rising ratios of CFP-Rac1:Venus-IQGAP1 were correlated with lower overall actin fluorescence, whereas the CFP-Cdc42:Venus-IQGAP1 ratio correlated with increased actin fluorescence at low ratios, but was neutral at higher ratios. The new FRET analyses also indicated that rising levels of mRFP1-Cdc42 or mRFP1-Rac1, respectively, promoted or suppressed the association of Teal-N-WASP with Venus-IQGAP1. These 3-color FRET assays further support our in vitro results about the role of IQGAP1, Rac1, and Cdc42 in actin nucleation, and the differential impact of Rac1 and Cdc42 on the association of N-WASP with IQGAP1. In addition, this study emphasizes the power of 3-color FRET as a systems biology strategy for simultaneous evaluation of multiple interacting proteins in individual live cells. © 2015 International Society for Advancement of Cytometry.

  5. Nuclear structure studies on medium-heavy mass nuclei using the method of nuclear resonance fluorescence; Kernstrukturuntersuchungen in mittelschweren Atomkernen mit der Methode der Kernresonanzfluoreszenz

    Zweidinger, Markus

    2016-06-22

    In the present work the dipole strength distribution in the stable even-even isotopes {sup 92}Zr and {sup 94}Zr is investigated. To excite the nuclei from the ground state to an excited state, real photons are used. This method is called Nuclear Resonance Fluorescence. The measurements were performed at two different setups. The first one is the Darmstadt High Intensity Photon Setup (DHIPS). At DHIPS the measurements yield information about the spin quantum number and the integrated cross section. The second part of the experiments took place at the High Intensity γ-ray Source (HIγS). Here, information about the parity quantum number and the averaged branching ratio of the excited state is accessible. In total, 105 dipole excited states in the nucleus {sup 92}Zr and 124 in the isotope {sup 94}Zr are observed, most of them for the first time. The extracted dipole strength distribution is investigated for the existence of the pygmy dipole resonance that was observed in neighboring nuclei. Furthermore, in previously performed experiments on the isotope {sup 90}Zr, the spin-flip M1 resonance was observed as well. Therefore, also the magnetic dipole strength is investigated. Further, by comparison with global systematics, the two-phonon state is identified. Additionally, the averaged branching ratio is compared to the results of theoretical calculations in the framework of the statistical model.

  6. Low-energy d-d excitations in MnO studied by resonant x-ray fluorescence spectroscopy

    Butorin, S.M.; Guo, J.; Magnuson, M.

    1997-01-01

    Resonant soft X-ray emission spectroscopy has been demonstrated to possess interesting abilities for studies of electronic structure in various systems, such as symmetry probing, alignment and polarization dependence, sensitivity to channel interference, etc. In the present abstract the authors focus on the feasibility of resonant soft X-ray emission to probe low energy excitations by means of resonant electronic X-ray Raman scattering. Resonant X-ray emission can be regarded as an inelastic scattering process where a system in the ground state is transferred to a low excited state via a virtual core excitation. The energy closeness to a core excitation of the exciting radiation enhances the (generally) low probability for inelastic scattering at these wavelengths. Therefore soft X-ray emission spectroscopy (in resonant electronic Raman mode) can be used to study low energy d-d excitations in transition metal systems. The involvement of the intermediate core state allows one to use the selection rules of X-ray emission, and the appearance of the elastically scattered line in the spectra provides the reference to the ground state

  7. Low-energy d-d excitations in MnO studied by resonant x-ray fluorescence spectroscopy

    Butorin, S.M.; Guo, J.; Magnuson, M. [Uppsala Univ. (Sweden)] [and others

    1997-04-01

    Resonant soft X-ray emission spectroscopy has been demonstrated to possess interesting abilities for studies of electronic structure in various systems, such as symmetry probing, alignment and polarization dependence, sensitivity to channel interference, etc. In the present abstract the authors focus on the feasibility of resonant soft X-ray emission to probe low energy excitations by means of resonant electronic X-ray Raman scattering. Resonant X-ray emission can be regarded as an inelastic scattering process where a system in the ground state is transferred to a low excited state via a virtual core excitation. The energy closeness to a core excitation of the exciting radiation enhances the (generally) low probability for inelastic scattering at these wavelengths. Therefore soft X-ray emission spectroscopy (in resonant electronic Raman mode) can be used to study low energy d-d excitations in transition metal systems. The involvement of the intermediate core state allows one to use the selection rules of X-ray emission, and the appearance of the elastically scattered line in the spectra provides the reference to the ground state.

  8. Simultaneous determination of reactive oxygen and nitrogen species in mitochondrial compartments of apoptotic HepG2 cells and PC12 cells based on microchip electrophoresis-laser-induced fluorescence.

    Chen, Zhenzhen; Li, Qingling; Sun, Qianqian; Chen, Hao; Wang, Xu; Li, Na; Yin, Miao; Xie, Yanxia; Li, Hongmin; Tang, Bo

    2012-06-05

    Determination of intracellular bioactive species will afford beneficial information related to cell metabolism, signal transduction, cell function, and disease treatment. In this study, the first application of a microchip electrophoresis-laser-induced fluorescence (MCE-LIF) method for concurrent determination of reactive oxygen species (ROS) and reactive nitrogen species (RNS), i.e., superoxide (O(2)(-•)) and nitric oxide (NO) in mitochondria, was developed using fluorescent probes 2-chloro-1,3-dibenzothiazolinecyclohexene (DBZTC) and 3-amino,4-aminomethyl-2',7'-difluorescein (DAF-FM), respectively. Potential interference of intracellular dehydroascorbic acid (DHA) and ascorbic acid (AA) for NO detection with DAF-FM was eliminated through oxidation of AA with the addition of ascorbate oxidase, followed by subsequent MCE separation. Fluorescent products of O(2)(-•) and NO, DBZTC oxide (DBO), and DAF-FM triazole (DAF-FMT) showed excellent baseline separation within 1 min with a running buffer of 40 mM Tris solution (pH 7.4) and a separating electric field of 500 V/cm. The levels of DBO and DAF-FMT in mitochondria isolated from normal HepG2 cells and PC12 cells were evaluated using this method. Furthermore, the changes of DBO and DAF-FMT levels in mitochondria isolated from apoptotic HepG2 cells and PC12 cells could also be detected. The current approach was proved to be simple, fast, reproducible, and efficient. Measurement of the two species with the method will be beneficial to understand ROS/RNS distinctive functions. In addition, it will provide new insights into the role that both species play in biological systems.

  9. The Value of 5-Aminolevulinic Acid in Low-grade Gliomas and High-grade Gliomas Lacking Glioblastoma Imaging Features: An Analysis Based on Fluorescence, Magnetic Resonance Imaging, 18F-Fluoroethyl Tyrosine Positron Emission Tomography, and Tumor Molecular Factors.

    Jaber, Mohammed; Wölfer, Johannes; Ewelt, Christian; Holling, Markus; Hasselblatt, Martin; Niederstadt, Thomas; Zoubi, Tarek; Weckesser, Matthias; Stummer, Walter

    2016-03-01

    Approximately 20% of grade II and most grade III gliomas fluoresce after 5-aminolevulinic acid (5-ALA) application. Conversely, approximately 30% of nonenhancing gliomas are actually high grade. The aim of this study was to identify preoperative factors (ie, age, enhancement, 18F-fluoroethyl tyrosine positron emission tomography [F-FET PET] uptake ratios) for predicting fluorescence in gliomas without typical glioblastomas imaging features and to determine whether fluorescence will allow prediction of tumor grade or molecular characteristics. Patients harboring gliomas without typical glioblastoma imaging features were given 5-ALA. Fluorescence was recorded intraoperatively, and biopsy specimens collected from fluorescing tissue. World Health Organization (WHO) grade, Ki-67/MIB-1 index, IDH1 (R132H) mutation status, O-methylguanine DNA methyltransferase (MGMT) promoter methylation status, and 1p/19q co-deletion status were assessed. Predictive factors for fluorescence were derived from preoperative magnetic resonance imaging and F-FET PET. Classification and regression tree analysis and receiver-operating-characteristic curves were generated for defining predictors. Of 166 tumors, 82 were diagnosed as WHO grade II, 76 as grade III, and 8 as glioblastomas grade IV. Contrast enhancement, tumor volume, and F-FET PET uptake ratio >1.85 predicted fluorescence. Fluorescence correlated with WHO grade (P fluorescing grade III gliomas was higher than in nonfluorescing tumors, whereas in fluorescing and nonfluorescing grade II tumors, no differences were noted. Age, tumor volume, and F-FET PET uptake are factors predicting 5-ALA-induced fluorescence in gliomas without typical glioblastoma imaging features. Fluorescence was associated with an increased Ki-67/MIB-1 index and high-grade pathology. Whether fluorescence in grade II gliomas identifies a subtype with worse prognosis remains to be determined.

  10. pH-Responsive Tumor-Targetable Theranostic Nanovectors Based on Core Crosslinked (CCL Micelles with Fluorescence and Magnetic Resonance (MR Dual Imaging Modalities and Drug Delivery Performance

    Sidan Tian

    2016-06-01

    Full Text Available The development of novel theranostic nanovectors is of particular interest in treating formidable diseases (e.g., cancers. Herein, we report a new tumor-targetable theranostic agent based on core crosslinked (CCL micelles, possessing tumor targetable moieties and fluorescence and magnetic resonance (MR dual imaging modalities. An azide-terminated diblock copolymer, N3-POEGMA-b-P(DPA-co-GMA, was synthesized via consecutive atom transfer radical polymerization (ATRP, where OEGMA, DPA, and GMA are oligo(ethylene glycolmethyl ether methacrylate, 2-(diisopropylaminoethyl methacrylate, and glycidyl methacrylate, respectively. The resulting diblock copolymer was further functionalized with DOTA(Gd (DOTA is 1,4,7,10-tetraazacyclododecane-1,4,7,10-tetrakisacetic acid or benzaldehyde moieties via copper(I-catalyzed alkyne-azide cycloaddition (CuAAC chemistry, resulting in the formation of DOTA(Gd-POEGMA-b-P(DPA-co-GMA and benzaldehyde-POEGMA-b-P(DPA-co-GMA copolymers. The resultant block copolymers co-assembled into mixed micelles at neutral pH in the presence of tetrakis[4-(2-mercaptoethoxyphenyl]ethylene (TPE-4SH, which underwent spontaneous crosslinking reactions with GMA residues embedded within the micellar cores, simultaneously switching on TPE fluorescence due to the restriction of intramolecular rotation. Moreover, camptothecin (CPT was encapsulated into the crosslinked cores at neutral pH, and tumor-targeting pH low insertion peptide (pHLIP, sequence: AEQNPIYWARYADWLFTTPLLLLDLALLVDADEGTCG moieties were attached to the coronas through the Schiff base chemistry, yielding a theranostic nanovector with fluorescence and MR dual imaging modalities and tumor-targeting capability. The nanovectors can be efficiently taken up by A549 cells, as monitored by TPE fluorescence. After internalization, intracellular acidic pH triggered the release of loaded CPT, killing cancer cells in a selective manner. On the other hand, the nanovectors labeled with DOTA

  11. Fluorescence and Magnetic Resonance Dual-Modality Imaging-Guided Photothermal and Photodynamic Dual-Therapy with Magnetic Porphyrin-Metal Organic Framework Nanocomposites

    Zhang, Hui; Li, Yu-Hao; Chen, Yang; Wang, Man-Man; Wang, Xue-Sheng; Yin, Xue-Bo

    2017-03-01

    Phototherapy shows some unique advantages in clinical application, such as remote controllability, improved selectivity, and low bio-toxicity, than chemotherapy. In order to improve the safety and therapeutic efficacy, imaging-guided therapy seems particularly important because it integrates visible information to speculate the distribution and metabolism of the probe. Here we prepare biocompatible core-shell nanocomposites for dual-modality imaging-guided photothermal and photodynamic dual-therapy by the in situ growth of porphyrin-metal organic framework (PMOF) on Fe3O4@C core. Fe3O4@C core was used as T2-weighted magnetic resonance (MR) imaging and photothermal therapy (PTT) agent. The optical properties of porphyrin were well remained in PMOF, and PMOF was therefore selected for photodynamic therapy (PDT) and fluorescence imaging. Fluorescence and MR dual-modality imaging-guided PTT and PDT dual-therapy was confirmed with tumour-bearing mice as model. The high tumour accumulation of Fe3O4@C@PMOF and controllable light excitation at the tumour site achieved efficient cancer therapy, but low toxicity was observed to the normal tissues. The results demonstrated that Fe3O4@C@PMOF was a promising dual-imaging guided PTT and PDT dual-therapy platform for tumour diagnosis and treatment with low cytotoxicity and negligible in vivo toxicity.

  12. Raman and fluorescence contributions to the resonant inelastic soft x-ray scattering on LaAlO3/SrTiO3 heterostructures

    Pfaff, F.; Fujiwara, H.; Berner, G.; Yamasaki, A.; Niwa, H.; Kiuchi, H.; Gloskovskii, A.; Drube, W.; Gabel, J.; Kirilmaz, O.; Sekiyama, A.; Miyawaki, J.; Harada, Y.; Suga, S.; Sing, M.; Claessen, R.

    2018-01-01

    We present a detailed study of the Ti 3 d carriers at the interface of LaAlO3/SrTiO3 heterostructures by high-resolution resonant inelastic soft x-ray scattering (RIXS), with special focus on the roles of overlayer thickness and oxygen vacancies. Our measurements show the existence of interfacial Ti 3 d electrons already below the critical thickness for conductivity. The (total) interface charge carrier density increases up to a LaAlO3 overlayer thickness of 6 unit cells before it levels out. Furthermore, we observe strong Ti 3 d charge carrier doping by oxygen vacancies. The RIXS data combined with photoelectron spectroscopy and transport measurements indicate the simultaneous presence of localized and itinerant charge carriers. At variance with previous interpretations, we show that in our excitation energy dependent RIXS measurements the amounts of localized and itinerant Ti 3 d electrons in the ground state do not scale with the intensities of the Raman and fluorescence peaks, respectively. Rather, we attribute the observation of either Raman components or fluorescence signal to the specific nature of the intermediate state reached in the RIXS excitation process.

  13. Enhancing molecular logic through modulation of temporal and spatial constraints with quantum dot-based systems that use fluorescent (Förster) resonance energy transfer

    Claussen, Jonathan C.; Algar, W. Russ; Hildebrandt, Niko; Susumu, Kimihiro; Ancona, Mario G.; Medintz, Igor L.

    2013-10-01

    Luminescent semiconductor nanocrystals or quantum dots (QDs) contain favorable photonic properties (e.g., resistance to photobleaching, size-tunable PL, and large effective Stokes shifts) that make them well-suited for fluorescence (Förster) resonance energy transfer (FRET) based applications including monitoring proteolytic activity, elucidating the effects of nanoparticles-mediated drug delivery, and analyzing the spatial and temporal dynamics of cellular biochemical processes. Herein, we demonstrate how unique considerations of temporal and spatial constraints can be used in conjunction with QD-FRET systems to open up new avenues of scientific discovery in information processing and molecular logic circuitry. For example, by conjugating both long lifetime luminescent terbium(III) complexes (Tb) and fluorescent dyes (A647) to a single QD, we can create multiple FRET lanes that change temporally as the QD acts as both an acceptor and donor at distinct time intervals. Such temporal FRET modulation creates multi-step FRET cascades that produce a wealth of unique photoluminescence (PL) spectra that are well-suited for the construction of a photonic alphabet and photonic logic circuits. These research advances in bio-based molecular logic open the door to future applications including multiplexed biosensing and drug delivery for disease diagnostics and treatment.

  14. Intramolecular ex vivo Fluorescence Resonance Energy Transfer (FRET of Dihydropyridine Receptor (DHPR β1a Subunit Reveals Conformational Change Induced by RYR1 in Mouse Skeletal Myotubes.

    Dipankar Bhattacharya

    Full Text Available The dihydropyridine receptor (DHPR β1a subunit is essential for skeletal muscle excitation-contraction coupling, but the structural organization of β1a as part of the macromolecular DHPR-ryanodine receptor type I (RyR1 complex is still debatable. We used fluorescence resonance energy transfer (FRET to probe proximity relationships within the β1a subunit in cultured skeletal myotubes lacking or expressing RyR1. The fluorescein biarsenical reagent FlAsH was used as the FRET acceptor, which exhibits fluorescence upon binding to specific tetracysteine motifs, and enhanced cyan fluorescent protein (CFP was used as the FRET donor. Ten β1a reporter constructs were generated by inserting the CCPGCC FlAsH binding motif into five positions probing the five domains of β1a with either carboxyl or amino terminal fused CFP. FRET efficiency was largest when CCPGCC was positioned next to CFP, and significant intramolecular FRET was observed for all constructs suggesting that in situ the β1a subunit has a relatively compact conformation in which the carboxyl and amino termini are not extended. Comparison of the FRET efficiency in wild type to that in dyspedic (lacking RyR1 myotubes revealed that in only one construct (H458 CCPGCC β1a -CFP FRET efficiency was specifically altered by the presence of RyR1. The present study reveals that the C-terminal of the β1a subunit changes conformation in the presence of RyR1 consistent with an interaction between the C-terminal of β1a and RyR1 in resting myotubes.

  15. Observation of resonance fluorescence and the Mollow triplet from a coherently driven site-controlled quantum dot

    Unsleber, Sebastian; Maier, Sebastian; McCutcheon, Dara

    2015-01-01

    -controlled semiconductor quantum dot to an external resonant laser field. For strong continuous-wave driving we observe the characteristic Mollow triplet and analyze the Rabi splitting and sideband widths as a function of driving strength and temperature. The sideband widths increase linearly with temperature...... and the square of the driving strength, which we explain via coupling of the exciton to longitudinal acoustic phonons. We also find an increase of the Rabi splitting with temperature, which indicates a temperature induced delocalization of the excitonic wave function resulting in an increase of the oscillator...... strength. Finally, we demonstrate coherent control of the exciton excited state population via pulsed resonant excitation and observe a damping of the Rabi oscillations with increasing pulse area, which is consistent with our exciton-photon coupling model. We believe that our work outlines the possibility...

  16. A Geant4-based Simulation to Evaluate the Feasibility of Using Nuclear Resonance Fluorescence (NRF) in Determining Atomic Compositions of Body Tissue in Cancer Diagnostics and Irradiation

    Gilbo, Yekaterina; Wijesooriya, Krishni; Liyanage, Nilanga

    2017-01-01

    Customarily applied in homeland security for identifying concealed explosives and chemical weapons, NRF (Nuclear Resonance Fluorescence) may have high potential in determining atomic compositions of body tissue. High energy photons incident on a target excite the target nuclei causing characteristic re-emission of resonance photons. As the nuclei of each isotope have well-defined excitation energies, NRF uniquely indicates the isotopic content of the target. NRF radiation corresponding to nuclear isotopes present in the human body is emitted during radiotherapy based on Bremsstrahlung photons generated in a linear electron accelerator. We have developed a Geant4 simulation in order to help assess NRF capabilities in detecting, mapping, and characterizing tumors. We have imported a digital phantom into the simulation using anatomical data linked to known chemical compositions of various tissues. Work is ongoing to implement the University of Virginia's cancer center treatment setup and patient geometry, and to collect and analyze the simulation's physics quantities to evaluate the potential of NRF for medical imaging applications. Preliminary results will be presented.

  17. Fluorescent scattering by molecules embedded in small particles

    1982-01-01

    Studies are reported in these areas: double resonance in fluorescent and Raman scattering; surface enhanced Raman scattering; fluorescence by molecules embedded in small particles; fluorescence by a liquid droplet; and fluorescence by conical pits in surfaces

  18. Complicated Fermi-type vibronic resonance: Untangling of the single-site quasi-line fluorescence excitation spectra of a methylated dibenzoporphin

    Arabei, S.M.; Kuzmitsky, V.A.; Solovyov, K.N.

    2008-01-01

    The quasi-line low-temperature (4.2 K) fluorescence excitation spectra of 2,3,12,13-tetramethyldibenzo[g,q]porphin introduced into an n-octane matrix have been measured in the range of the S 2 0 electronic transition at selective fluorescence monitoring for the two main types of impurity centers (sites). A characteristic feature of these spectra is that a conglomerate of quasi-lines - a structured complex band - is observed instead of one 0-0 quasi-line of the S 2 0 transition. In this band, the intensity distributions for the two main sites considerably differ from each other. The occurrence of such conglomerates is interpreted as a result of nonadiabatic vibrational-electronic interaction between the vibronic S 2 and S 1 states (the complex vibronic analogue of the Fermi resonance). The frequencies and intensities of individual transitions determined from the deconvolution of complex conglomerates are used as the initial data for solving the inverse spectroscopic problem: the determination of the unperturbed electronic and vibrational levels of states involved in the resonance and the vibronic-interaction matrix elements between them. This problem is solved with a method developed previously. The experimental results and their analysis are compared to the analogous data obtained earlier for meso-tetraazaporphin and meso-tetrapropylporphin. The energy intervals between the S 2 and S 1 electronic levels (ΔE S 2 S 1 ) of the two main types of impurity centers formed by molecules of a given porphyrin in the crystal matrix are found to significantly differ from each other, the values of this difference (δΔE S 2 S 1 ) being considerably greater for tetramethyldibenzoporphin, δΔE S 2 S 1 =228cm -1 , than for the two other porphyrins. At the same time, the energies of the unperturbed vibrational states of the S 1 electronic level participating in the resonance are very close to each other for these two sites

  19. Coarse architecture of the transient receptor potential vanilloid 1 (TRPV1) ion channel determined by fluorescence resonance energy transfer.

    De-la-Rosa, Víctor; Rangel-Yescas, Gisela E; Ladrón-de-Guevara, Ernesto; Rosenbaum, Tamara; Islas, León D

    2013-10-11

    The transient receptor potential vanilloid 1 ion channel is responsible for the perception of high temperatures and low extracellular pH, and it is also involved in the response to some pungent compounds. Importantly, it is also associated with the perception of pain and noxious stimuli. Here, we attempt to discern the molecular organization and location of the N and C termini of the transient receptor potential vanilloid 1 ion channel by measuring FRET between genetically attached enhanced yellow and cyan fluorescent protein to the N or C terminus of the channel protein, expressed in transfected HEK 293 cells or Xenopus laevis oocytes. The static measurements of the domain organization were mapped into an available cryo-electron microscopy density of the channel with good agreement. These measurements also provide novel insights into the organization of terminal domains and their proximity to the plasma membrane.

  20. Depth profiles of pulmonary surfactant protein B in phosphatidylcholine bilayers, studied by fluorescence and electron spin resonance spectroscopy

    Cruz, A; Casals, C; Plasencia, I

    1998-01-01

    Pulmonary surfactant-associated protein B (SP-B) has been isolated from porcine lungs and reconstituted in bilayers of dipalmitoylphosphatidylcholine (DPPC) or egg yolk phosphatidylcholine (PC) to characterize the extent of insertion of the protein into phospholipid bilayers. The parameters...... for the interaction of SP-B with DPPC or PC using different reconstitution protocols have been estimated from the changes induced in the fluorescence emission spectrum of the single protein tryptophan. All the different reconstituted SP-B-phospholipid preparations studied had similar Kd values for the binding...... that there are significant differences in the extent of insertion of the protein, depending on the method of reconstitution. SP-B reconstituted from lipid/protein mixtures in organic solvents is inserted more deeply in PC or DPPC bilayers than the protein reconstituted by addition to preformed phospholipid vesicles...

  1. Graphene and graphene-like two-denominational materials based fluorescence resonance energy transfer (FRET) assays for biological applications.

    Tian, Feng; Lyu, Jing; Shi, Jingyu; Yang, Mo

    2017-03-15

    In the past decades, Förster resonance energy transfer (FRET) has been applied in many biological applications to reveal the biological information at the nanoscale. Recently, graphene and graphene-like two-dimensional (2D) nanomaterials started to be used in FRET assays as donors or acceptors including graphene oxide (GO), graphene quantum dot (GQD), graphitic-carbon nitride nanosheets (g-C 3 N 4 ) and transition metal dichalcogenides (e.g. MoS 2 , MnO 2, and WS 2 ). Due to the remarkable properties such as large surface to volume ratio, tunable energy band, photoluminescence and excellent biocompatibility, these 2D nanomaterials based FRET assays have shown great potential in various biological applications. This review summarizes the recent development of graphene and graphene-like 2D nanomaterials based FRET assays in applications of biosensing, bioimaging, and drug delivery monitoring. Copyright © 2016 Elsevier B.V. All rights reserved.

  2. A fluorescence resonance energy transfer-based approach for investigating late endosome–lysosome retrograde fusion events

    Kaufmann, A.M.; Goldman, S.D.B.; Krise, J.P.

    2009-01-01

    Traditionally, lysosomes have been considered to be a terminal endocytic compartment. Recent studies suggest that lysosomes are quite dynamic, being able to fuse with other late endocytic compartments as well as with the plasma membrane. Here we describe a quantitative fluorescence energy transfer (FRET)-based method for assessing rates of retrograde fusion between terminal lysosomes and late endosomes in living cells. Late endosomes were specifically labeled with 800-nm latex beads that were conjugated with streptavidin and Alexa Fluor 555 (FRET donor). Terminal lysosomes were specifically labeled with 10,000-MW dextran polymers conjugated with biotin and Alexa Fluor 647 (FRET acceptor). Following late endosome–lysosome fusion, the strong binding affinity between streptavidin and biotin brought the donor and acceptor fluorophore molecules into close proximity, thereby facilitating the appearance of a FRET emission signal. Because apparent size restrictions in the endocytic pathway do not permit endocytosed latex beads from reaching terminal lysosomes in an anterograde fashion, the appearance of the FRET signal is consistent with retrograde transport of lysosomal cargo back to late endosomes. We assessed the efficiency of this transport step in fibroblasts affected by different lysosome storage disorders—Niemann–Pick type C, mucolipidosis type IV, and Sandhoff’s disease, all of which have a similar lysosomal lipid accumulation phenotype. We report here, for the first time, that these disorders can be distinguished by their rate of transfer of lysosome cargos to late endosomes, and we discuss the implications of these findings for developing new therapeutic strategies. PMID:19109922

  3. A fluorescence resonance energy transfer-based approach for investigating late endosome-lysosome retrograde fusion events.

    Kaufmann, A M; Goldman, S D B; Krise, J P

    2009-03-01

    Traditionally, lysosomes have been considered to be a terminal endocytic compartment. Recent studies suggest that lysosomes are quite dynamic, being able to fuse with other late endocytic compartments as well as with the plasma membrane. Here we describe a quantitative fluorescence energy transfer (FRET)-based method for assessing rates of retrograde fusion between terminal lysosomes and late endosomes in living cells. Late endosomes were specifically labeled with 800-nm latex beads that were conjugated with streptavidin and Alexa Fluor 555 (FRET donor). Terminal lysosomes were specifically labeled with 10,000-MW dextran polymers conjugated with biotin and Alexa Fluor 647 (FRET acceptor). Following late endosome-lysosome fusion, the strong binding affinity between streptavidin and biotin brought the donor and acceptor fluorophore molecules into close proximity, thereby facilitating the appearance of a FRET emission signal. Because apparent size restrictions in the endocytic pathway do not permit endocytosed latex beads from reaching terminal lysosomes in an anterograde fashion, the appearance of the FRET signal is consistent with retrograde transport of lysosomal cargo back to late endosomes. We assessed the efficiency of this transport step in fibroblasts affected by different lysosome storage disorders-Niemann-Pick type C, mucolipidosis type IV, and Sandhoff's disease, all of which have a similar lysosomal lipid accumulation phenotype. We report here, for the first time, that these disorders can be distinguished by their rate of transfer of lysosome cargos to late endosomes, and we discuss the implications of these findings for developing new therapeutic strategies.

  4. Optical-optical double resonance, laser induced fluorescence, and revision of the signs of the spin-spin constants of the boron carbide (BC) free radical

    Sunahori, Fumie X.; Nagarajan, Ramya; Clouthier, Dennis J.

    2015-12-01

    The cold boron carbide free radical (BC X 4Σ-) has been produced in a pulsed discharge free jet expansion using a precursor mixture of trimethylborane in high pressure argon. High resolution laser induced fluorescence spectra have been obtained for the B 4Σ--X 4Σ- and E 4Π-X 4Σ- band systems of both 11BC and 10BC. An optical-optical double resonance (OODR) scheme was implemented to study the finer details of both band systems. This involved pumping a single rotational level of the B state with one laser and then recording the various allowed transitions from the intermediate B state to the final E state with a second laser by monitoring the subsequent E-X ultraviolet fluorescence. In this fashion, we were able to prove unambiguously that, contrary to previous studies, the spin-spin constant λ is negative in the ground state and positive in the B 4Σ- excited state. It has been shown that λ″ expected based on a semiempirical second order perturbation theory calculation of the magnitude of the spin-spin constant. The OODR spectra have also been used to validate our assignments of the complex and badly overlapped E 4Π-X 4Σ- 0-0 and 1-0 bands of 11BC. The E-X 0-0 band of 10BC was found to be severely perturbed. The ground state main electron configuration is …3σ24σ25σ11π22π0 and the derived bond lengths show that there is a 0.03 Å contraction in the B state, due to the promotion of an electron from the 4σ antibonding orbital to the 5σ bonding orbital. In contrast, the bond length elongates by 0.15 Å in the E state, a result of promoting an electron from the 5σ bonding orbital to the 2π antibonding orbitals.

  5. Gastrin-releasing peptide receptor-targeted gadolinium oxide-based multifunctional nanoparticles for dual magnetic resonance/fluorescent molecular imaging of prostate cancer

    Cui DT

    2017-09-01

    Full Text Available Danting Cui,1 Xiaodan Lu,1 Chenggong Yan,1 Xiang Liu,1 Meirong Hou,1 Qi Xia,2 Yikai Xu,1 Ruiyuan Liu2,3 1Department of Medical Imaging Center, Nanfang Hospital, Southern Medical University, Guangzhou, People’s Republic of China; 2School of Pharmaceutical Sciences, Southern Medical University, Guangzhou, People’s Republic of China; 3School of Biomedical Engineering, Southern Medical University, Guangzhou, People’s Republic of China Abstract: Bombesin (BBN, an analog of gastrin-releasing peptide (GRP, specifically binds to GRP receptors, which are overexpressed in human prostate cancer (PC. Here, we synthesized a BBN-modified gadolinium oxide (Gd2O3 nanoprobe containing fluorescein (Gd2O3-5(6-carboxyfluorescein [FI]-polyethylene glycol [PEG]-BBN for targeted magnetic resonance (MR/optical dual-modality imaging of PC. The Gd2O3-FI-PEG-BBN nanoparticles exhibited a relatively uniform particle size with an average diameter of 52.3 nm and spherical morphology as depicted by transmission electron microscopy. The longitudinal relaxivity (r1 of Gd2O3-FI-PEG-BBN (r1 =4.23 mM–1s–1 is comparable to that of clinically used Magnevist (Gd-DTPA. Fluorescence microscopy and in vitro cellular MRI demonstrated GRP receptor-specific and enhanced cellular uptake of the Gd2O3-FI-PEG-BBN in PC-3 tumor cells. Moreover, Gd2O3-FI-PEG-BBN showed more remarkable contrast enhancement than the corresponding nontargeted Gd2O3-FI-PEG according to in vivo MRI and fluorescent imaging. Tumor immunohistochemical analysis further demonstrated improved accumulation of the targeted nanoprobe in tumors. BBN-conjugated Gd2O3 may be a promising nanoplatform for simultaneous GRP receptor-targeted molecular cancer diagnosis and antitumor drug delivery in future clinical applications. Keywords: magnetic resonance imaging, gadolinium oxide, bombesin, gastrin-releasing peptide receptor, molecular imaging

  6. An in vitro tag-and-modify protein sample generation method for single-molecule fluorescence resonance energy transfer.

    Hamadani, Kambiz M; Howe, Jesse; Jensen, Madeleine K; Wu, Peng; Cate, Jamie H D; Marqusee, Susan

    2017-09-22

    Biomolecular systems exhibit many dynamic and biologically relevant properties, such as conformational fluctuations, multistep catalysis, transient interactions, folding, and allosteric structural transitions. These properties are challenging to detect and engineer using standard ensemble-based techniques. To address this drawback, single-molecule methods offer a way to access conformational distributions, transient states, and asynchronous dynamics inaccessible to these standard techniques. Fluorescence-based single-molecule approaches are parallelizable and compatible with multiplexed detection; to date, however, they have remained limited to serial screens of small protein libraries. This stems from the current absence of methods for generating either individual dual-labeled protein samples at high throughputs or protein libraries compatible with multiplexed screening platforms. Here, we demonstrate that by combining purified and reconstituted in vitro translation, quantitative unnatural amino acid incorporation via AUG codon reassignment, and copper-catalyzed azide-alkyne cycloaddition, we can overcome these challenges for target proteins that are, or can be, methionine-depleted. We present an in vitro parallelizable approach that does not require laborious target-specific purification to generate dual-labeled proteins and ribosome-nascent chain libraries suitable for single-molecule FRET-based conformational phenotyping. We demonstrate the power of this approach by tracking the effects of mutations, C-terminal extensions, and ribosomal tethering on the structure and stability of three protein model systems: barnase, spectrin, and T4 lysozyme. Importantly, dual-labeled ribosome-nascent chain libraries enable single-molecule co-localization of genotypes with phenotypes, are well suited for multiplexed single-molecule screening of protein libraries, and should enable the in vitro directed evolution of proteins with designer single-molecule conformational

  7. Multiplexed interfacial transduction of nucleic acid hybridization using a single color of immobilized quantum dot donor and two acceptors in fluorescence resonance energy transfer.

    Algar, W Russ; Krull, Ulrich J

    2010-01-01

    A multiplexed solid-phase assay for the detection of nucleic acid hybridization was developed on the basis of a single color of immobilized CdSe/ZnS quantum dot (QD) as a donor in fluorescence resonance energy transfer (FRET). This work demonstrated that two channels of detection did not necessitate two different QD donors. Two probe oligonucleotides were coimmobilized on optical fibers modified with QDs, and a sandwich assay was used to associate the acceptor dyes with interfacial hybridization events without target labeling. FRET-sensitized acceptor emission provided an analytical signal that was concentration dependent down to 10 nM. Changes in the ratio of coimmobilized probe oligonucleotides were found to yield linear changes in the relative amounts of acceptor emission. These changes were compared to previous studies that used mixed films of two QD donors for two detection channels. The analysis indicated that probe dilution effects were primarily driven by changes in acceptor number density and that QD dilution effects or changes in mean donor-acceptor distance were secondary. Hybridization kinetics were found to be consistent between different ratios of coimmobilized probes, suggesting that hybridization in this type of system occurred via the accepted model for solid-phase hybridization, where adsorption and then diffusion at the solid interface drove hybridization.

  8. Interfacial chemistry and the design of solid-phase nucleic acid hybridization assays using immobilized quantum dots as donors in fluorescence resonance energy transfer.

    Algar, W Russ; Krull, Ulrich J

    2011-01-01

    The use of quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET) offer several advantages for the development of multiplexed solid-phase QD-FRET nucleic acid hybridization assays. Designs for multiplexing have been demonstrated, but important challenges remain in the optimization of these systems. In this work, we identify several strategies based on the design of interfacial chemistry for improving sensitivity, obtaining lower limits of detection (LOD) and enabling the regeneration and reuse of solid-phase QD-FRET hybridization assays. FRET-sensitized emission from acceptor dyes associated with hybridization events at immobilized QD donors provides the analytical signal in these assays. The minimization of active sensing area reduces background from QD donor PL and allows the resolution of smaller amounts of acceptor emission, thus lowering the LOD. The association of multiple acceptor dyes with each hybridization event can enhance FRET efficiency, thereby improving sensitivity. Many previous studies have used interfacial protein layers to generate selectivity; however, transient destabilization of these layers is shown to prevent efficient regeneration. To this end, we report a protein-free interfacial chemistry and demonstrate the specific detection of as little as 2 pmol of target, as well as an improved capacity for regeneration.

  9. Determination of the electromagnetic dipole strength distribution in medium-heavy atomic nuclei by means of nuclear resonance fluorescence; Bestimmung der elektromagnetischen Dipolstaerkeverteilung in mittelschweren Atomkernen mittels Kernresonanzfluoreszenz

    Massarczyk, Ralph Jens

    2011-01-17

    During the last hundred years several models were developed to describe the configuration of nuclei. These models have to make predictions, which should be comparable with experiments. As a standard type of experiment the nuclear resonance fluorescence was established. A nucleus is excited by irradiation with photons. By emitting one or more photons the nucleus decays back to the ground state. With this method it is possible to measure energy levels and to determine the strength of their excitation. A continuum of unresolved peaks gives additional strength. The existing setup at the linear electron accelerator ELBE of the Forschungszentrum Dresden-Rossendorf uses bremsstrahlung, produced as a secondary beam in a thin Niobium foil. During the years 2008/09 experiments on the nuclei of {sup 86}Kr and {sup 136}Ba took place there. In this work they will be analyzed. Photon flux and efficiency determination have been done as well as simulations on detector response and non-nuclear scattered background events. For this purpose the GEANT4 package was used. Finally the resulting cross sections were corrected for branching and feeding.

  10. Detection of norovirus virus-like particles using a surface plasmon resonance-assisted fluoroimmunosensor optimized for quantum dot fluorescent labels.

    Ashiba, Hiroki; Sugiyama, Yuki; Wang, Xiaomin; Shirato, Haruko; Higo-Moriguchi, Kyoko; Taniguchi, Koki; Ohki, Yoshimichi; Fujimaki, Makoto

    2017-07-15

    A highly sensitive biosensor to detect norovirus in environment is desired to prevent the spread of infection. In this study, we investigated a design of surface plasmon resonance (SPR)-assisted fluoroimmunosensor to increase its sensitivity and performed detection of norovirus virus-like particles (VLPs). A quantum dot fluorescent dye was employed because of its large Stokes shift. The sensor design was optimized for the CdSe-ZnS-based quantum dots. The optimal design was applied to a simple SPR-assisted fluoroimmunosensor that uses a sensor chip equipped with a V-shaped trench. Excitation efficiency of the quantum dots, degree of electric field enhancement by SPR, and intensity of autofluorescence of a substrate of the sensor chip were theoretically and experimentally evaluated to maximize the signal-to-noise ratio. As the result, an excitation wavelength of 390nm was selected to excite SPR on an Al film of the sensor chip. The sandwich assay of norovirus VLPs was performed using the designed sensor. Minimum detectable concentration of 0.01ng/mL, which corresponds to 100 virus-like particles included in the detection region of the V-trench, was demonstrated. Copyright © 2016 The Authors. Published by Elsevier B.V. All rights reserved.

  11. Fluorescence resonance energy transfer between ZnSe ZnS quantum dots and bovine serum albumin in bioaffinity assays of anticancer drugs

    Shu, Chang; Ding, Li; Zhong, Wenying

    2014-10-01

    In the current work, using ZnSe ZnS quantum dots (QDs) as representative nanoparticles, the affinities of seven anticancer drugs for bovine serum albumin (BSA) were studied using fluorescence resonance energy transfer (FRET). The FRET efficiency of BSA-QD conjugates can reach as high as 24.87% by electrostatic interaction. The higher binding constant (3.63 × 107 L mol-1) and number of binding sites (1.75) between ZnSe ZnS QDs and BSA demonstrated that the QDs could easily associate to plasma proteins and enhance the transport efficacy of drugs. The magnitude of binding constants (103-106 L mol-1), in the presence of QDs, was between drugs-BSA and drugs-QDs in agreement with common affinities of drugs for serum albumins (104-106 L mol-1) in vivo. ZnSe ZnS QDs significantly increased the affinities for BSA of Vorinostat (SAHA), Docetaxel (DOC), Carmustine (BCNU), Doxorubicin (Dox) and 10-Hydroxycamptothecin (HCPT). However, they slightly reduced the affinities of Vincristine (VCR) and Methotrexate (MTX) for BSA. The recent work will not only provide useful information for appropriately understanding the binding affinity and binding mechanism at the molecular level, but also illustrate the ZnSe ZnS QDs are perfect candidates for nanoscal drug delivery system (DDS).

  12. Cy5.5 conjugated MnO nanoparticles for magnetic resonance/near-infrared fluorescence dual-modal imaging of brain gliomas.

    Chen, Ning; Shao, Chen; Li, Shuai; Wang, Zihao; Qu, Yanming; Gu, Wei; Yu, Chunjiang; Ye, Ling

    2015-11-01

    The fusion of molecular and anatomical modalities facilitates more reliable and accurate detection of tumors. Herein, we prepared the PEG-Cy5.5 conjugated MnO nanoparticles (MnO-PEG-Cy5.5 NPs) with magnetic resonance (MR) and near-infrared fluorescence (NIRF) imaging modalities. The applicability of MnO-PEG-Cy5.5 NPs as a dual-modal (MR/NIRF) imaging nanoprobe for the detection of brain gliomas was investigated. In vivo MR contrast enhancement of the MnO-PEG-Cy5.5 nanoprobe in the tumor region was demonstrated. Meanwhile, whole-body NIRF imaging of glioma bearing nude mouse exhibited distinct tumor localization upon injection of MnO-PEG-Cy5.5 NPs. Moreover, ex vivo CLSM imaging of the brain slice hosting glioma indicated the preferential accumulation of MnO-PEG-Cy5.5 NPs in the glioma region. Our results therefore demonstrated the potential of MnO-PEG-Cy5.5 NPs as a dual-modal (MR/NIRF) imaging nanoprobe in improving the diagnostic efficacy by simultaneously providing anatomical information from deep inside the body and more sensitive information at the cellular level. Copyright © 2015 Elsevier Inc. All rights reserved.

  13. In Vivo Dual-Modality Fluorescence and Magnetic Resonance Imaging-Guided Lymph Node Mapping with Good Biocompatibility Manganese Oxide Nanoparticles

    Yonghua Zhan

    2017-12-01

    Full Text Available Multifunctional manganese oxide nanoparticles (NPs with impressive enhanced T1 contrast ability show great promise in biomedical diagnosis. Herein, we developed a dual-modality imaging agent system based on polyethylene glycol (PEG-coated manganese oxide NPs conjugated with organic dye (Cy7.5, which functions as a fluorescence imaging (FI agent as well as a magnetic resonance imaging (MRI imaging agent. The formed Mn3O4@PEG-Cy7.5 NPs with the size of ~10 nm exhibit good colloidal stability in different physiological media. Serial FI and MRI studies that non-invasively assessed the bio-distribution pattern and the feasibility for in vivo dual-modality imaging-guided lymph node mapping have been investigated. In addition, histological and biochemical analyses exhibited low toxicity even at a dose of 20 mg/kg in vivo. Since Mn3O4@PEG-Cy7.5 NPs exhibited desirable properties as imaging agents and good biocompatibility, this work offers a robust, safe, and accurate diagnostic platform based on manganese oxide NPs for tumor metastasis diagnosis.

  14. Study on the interaction between albendazole and eosin Y by fluorescence, resonance Rayleigh scattering and frequency doubling scattering spectra and their analytical applications

    Tian, Fengling; Huang, Wei; Yang, Jidong; Li, Qin

    In pH 3.25-3.35 Britton-Robinson (BR) buffer solution, albendazole (ABZ) could react with eosin Y (EY) to form a 1:1 ion-association complex, which not only results in the quenching of fluorescence, but also resulted in the great enhancement of resonance Rayleigh scattering (RRS) and frequency doubling scattering (FDS). Furthermore, a new RRS spectrum will appear, and the maximum RRS wavelength was located at about 356 nm. The detection limit for ABZ were 21.51 ng mL-1 for the fluorophotometry, 6.93 ng mL-1 for the RRS method and 12.89 ng mL-1 for the FDS method. Among them, the RRS method had the highest sensitivity. The experimental conditions were optimized and effects of coexisting substances were evaluated. Meanwhile, the influences of coexisting substances were tested. The methods have been successfully applied to the determination of ABZ in capsules and human urine samples. The composition and structure of the ion-association complex and the reaction mechanism were discussed.

  15. Lateral distribution of NBD-PC fluorescent lipid analogs in membranes probed by molecular dynamics-assisted analysis of Förster Resonance Energy Transfer (FRET) and fluorescence quenching.

    Loura, Luís M S

    2012-11-08

    Förster resonance energy transfer (FRET) is a powerful tool used for many problems in membrane biophysics, including characterization of the lateral distribution of lipid components and other species of interest. However, quantitative analysis of FRET data with a topological model requires adequate choices for the values of several input parameters, some of which are difficult to obtain experimentally in an independent manner. For this purpose, atomistic molecular dynamics (MD) simulations can be potentially useful as they provide direct detailed information on transverse probe localization, relative probe orientation, and membrane surface area, all of which are required for analysis of FRET data. This is illustrated here for the FRET pairs involving 1,6-diphenylhexatriene (DPH) as donor and either 1-palmitoyl,2-(6-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino] hexanoyl)- sn-glycero-3-phosphocholine (C6-NBD-PC) or 1-palmitoyl,2-(12-[N-(7-nitrobenz-2-oxa-1,3-diazol-4-yl)amino]dodecanoyl)-sn-glycero-3-phosphocholine (C12-NBD-PC) as acceptors, in fluid vesicles of 1,2-dipalmitoyl-sn-3-glycerophosphocholine (DPPC, 50 °C). Incorporation of results from MD simulations improves the statistical quality of model fitting to the experimental FRET data. Furthermore, the decay of DPH in the presence of moderate amounts of C12-NBD-PC (>0.4 mol%) is consistent with non-random lateral distribution of the latter, at variance with C6-NBD-PC, for which aggregation is ruled out up to 2.5 mol% concentration. These conclusions are supported by analysis of NBD-PC fluorescence self-quenching. Implications regarding the relative utility of these probes in membrane studies are discussed.

  16. Application of the laser induced fluorescence to the investigation of highly magnetized plasmas, heated by ion cyclotron resonance; Fluorescence induite par laser sur des plasmas fortement magnetises, chauffes par resonnance cyclotron ionique

    Pailloux, A. [CEA Centre d`Etudes de Saclay, 91 - Gif-sur-Yvette (France). Dept. des Procedes d`Enrichissement]|[Universite Louis Pasteur, 67 - Strasbourg (France)

    1997-12-31

    This work has been achieved in the frame of isotopic separation studies by in cyclotron resonance. For this purpose, in a highly magnetized (2 to 3 Tesla) and non-collisional (10{sup 12} ions/cm{sup 3}) plasma, composed of metallic ions, a wave near the ion cyclotron frequency is thrown in order to heat selectively a given species. A laser induced fluorescence (LIP) has been developed on barium and gadolinium plasmas. The Larmor gyration of ions greatly modifies the interaction, which has been modelled through the time-dependent Schroedinger equation. The obtained excitation probably has been integrated over all the ions excited in the measurement volume in order to check that the LIF still leads to the distribution function of ion velocities. The influence of the Larmor motion of ions on the spectral distribution of LIF has been derived both theoretically and experimentally. The LIF diagnostics has been achieved with a dye O`ring laser. The barium ion has been excited on the transition 6142 angstrom, using rhodamine 6G dye, and the gadolinium ion on the pseudo-triplet 3861 angstrom, using exalite dye. Data treatment has been developed taking into account the Zeeman effect and the different heating of isotopes. The ionic temperature (from 1 eV to some hundreds eV) has been measured as a function of radiofrequency heating. Our experimental results are in good agreement with the selective heating theory. Also, the ion velocity distribution function has been found locally Maxwellian. And the behaviour of the plasma has been studied as a function of control parameters of the plasma source. (author) 62 refs.

  17. Study of the dispersion phenomena connected with the absorption by recoilless nuclear resonance fluorescence; Etude des phenomenes de dispersion lies a l'absorption resonnante sans recul des noyaux atomiques

    Imbert, P [Commissariat a l' Energie Atomique, Saclay (France). Centre d' Etudes Nucleaires

    1965-12-01

    In nuclear resonance fluorescence as in the optical field abnormal dispersion curves are related to the absorption lines. It is possible, by using quadrupolar or magnetic splitting of the line in the case of recoilless resonance fluorescence (Moessbauer effect) to obtain differential dispersion effects between the two orthogonal linear or the two inverse circular components of the incident gamma radiation. These effects induce bi-refraction phenomena or Faraday rotation on the gamma beam, which have been studied on Fe-57 enriched absorbers. (author) [French] Comme dans le domaine optique, aux raies d'absorption de fluorescence resonnante des noyaux atomiques sont associees des courbes de dispersion anormale. Les decompositions des raies d'absorption de fluorescence resonnante sans recul (raies Moessbauer) par couplage quadrupolaire ou effet Zeeman permettent d'obtenir des effets dispersifs differentiels entre composantes lineaires orthogonales ou circulaires inverses du rayonnement gamma incident. Ces effets se traduisent par des phenomenes de birefringence ou de rotation Faraday, qui ont pu etre etudies sur des milieux enrichis en fer-57. (auteur)

  18. Optical-optical double resonance, laser induced fluorescence, and revision of the signs of the spin-spin constants of the boron carbide (BC) free radical

    Sunahori, Fumie X. [Department of Chemistry and Physics, Franklin College, Franklin, Indiana 46131 (United States); Nagarajan, Ramya; Clouthier, Dennis J., E-mail: dclaser@uky.edu [Department of Chemistry, University of Kentucky, Lexington, Kentucky 40506-0055 (United States)

    2015-12-14

    The cold boron carbide free radical (BC X {sup 4}Σ{sup −}) has been produced in a pulsed discharge free jet expansion using a precursor mixture of trimethylborane in high pressure argon. High resolution laser induced fluorescence spectra have been obtained for the B {sup 4}Σ{sup −}–X {sup 4}Σ{sup −} and E {sup 4}Π–X {sup 4}Σ{sup −} band systems of both {sup 11}BC and {sup 10}BC. An optical-optical double resonance (OODR) scheme was implemented to study the finer details of both band systems. This involved pumping a single rotational level of the B state with one laser and then recording the various allowed transitions from the intermediate B state to the final E state with a second laser by monitoring the subsequent E–X ultraviolet fluorescence. In this fashion, we were able to prove unambiguously that, contrary to previous studies, the spin-spin constant λ is negative in the ground state and positive in the B {sup 4}Σ{sup −} excited state. It has been shown that λ″ < 0 is in fact expected based on a semiempirical second order perturbation theory calculation of the magnitude of the spin-spin constant. The OODR spectra have also been used to validate our assignments of the complex and badly overlapped E {sup 4}Π–X {sup 4}Σ{sup −} 0-0 and 1-0 bands of {sup 11}BC. The E–X 0-0 band of {sup 10}BC was found to be severely perturbed. The ground state main electron configuration is …3σ{sup 2}4σ{sup 2}5σ{sup 1}1π{sup 2}2π{sup 0} and the derived bond lengths show that there is a 0.03 Å contraction in the B state, due to the promotion of an electron from the 4σ antibonding orbital to the 5σ bonding orbital. In contrast, the bond length elongates by 0.15 Å in the E state, a result of promoting an electron from the 5σ bonding orbital to the 2π antibonding orbitals.

  19. The fluorescence and resonance Rayleigh scattering spectra study on the interactions of palladium (II)-Nootropic chelate with Congo red and their analytical applications

    Chen, Fang; Peng, Jingdong; Liu, Shaopu; Peng, Huanjun; Pan, Ziyu; Bu, Lingli; Xiao, Huan; Zhang, Ruiwen

    2017-04-01

    A highly sensitive detection approach of resonance Rayleigh scattering spectra (RRS) is firstly applied to analyzing nootropic drugs including piracetam (PIR) and oxiracetam (OXI). In HCl-NaAc buffer solution (pH = 3.0), the OXI chelated with palladium (II) to form the chelate cation [Pd2·OXI]2 +, and then reacted with Congo red (CGR) by virtue of electrostatic attraction and hydrophobic force to form binary complex [Pd2·OXI]. CGR2, which could result in the great enhancement of RRS. The resonance Rayleigh scattering signal was recorded at λex = λem = 375 nm. This mixture complex not only has higher RRS, but also makes contribution to significant increase of fluorescence, and the same phenomena also were discovered in PIR. The enhanced RRS intensity is in proportion to the PIR and OXI concentration in the range of 0.03-3.0 μg mL- 1, and the detection limit (DL) of RRS method for PIR and OXI is 2.3 ng mL- 1 and 9.7 ng mL- 1. In addition, the DL of fluorescence method for PIR and OXI is 8.4 μg mL- 1 and 19.5 μg mL- 1. Obviously, the RRS is the highly sensitive method, and the recoveries of the two kinds of nootropic drugs were range from 100.4% to 101.8.0% with RSD (n = 5) from 1.1% to 3.1% by RRS method. This paper not only investigated the optimum conditions for detecting nootropics with using RRS method, but also focused on the reasons for enhancing RRS intensity and the reaction mechanism, which in order to firm and contract the resultant. Finally, The RRS method has been applied to detect nootropic drugs in human urine samples with satisfactory results. Fig. S2. The effect of ionic strength: Pd (II)-CGR system (curve a); Pd (II)-OXI-CGR system (curve b); Pd (II)-PIR- CGR system (curve c). Pd (II): 2.0 × 10- 4 mol L- 1; CGR: 1.0 × 10- 5 mol L- 1; OXI: 1.5 μg mL- 1; PIR: 2 μg mL- 1; NaCl: 1 mol L- 1. Fig. S3. The effect of time: Pd (II)-OXI-CGR system (curve a); Pd (II)-PIR-CGR system (curve b). Pd (II): 2.0 × 10- 4 mol L- 1; CGR: 1.0 × 10- 5 mol L- 1

  20. On-chip multiplexed solid-phase nucleic acid hybridization assay using spatial profiles of immobilized quantum dots and fluorescence resonance energy transfer.

    Noor, M Omair; Tavares, Anthony J; Krull, Ulrich J

    2013-07-25

    A microfluidic based solid-phase assay for the multiplexed detection of nucleic acid hybridization using quantum dot (QD) mediated fluorescence resonance energy transfer (FRET) is described herein. The glass surface of hybrid glass-polydimethylsiloxane (PDMS) microfluidic channels was chemically modified to assemble the biorecognition interface. Multiplexing was demonstrated using a detection system that was comprised of two colors of immobilized semi-conductor QDs and two different oligonucleotide probe sequences. Green-emitting and red-emitting QDs were paired with Cy3 and Alexa Fluor 647 (A647) labeled oligonucleotides, respectively. The QDs served as energy donors for the transduction of dye labeled oligonucleotide targets. The in-channel assembly of the biorecognition interface and the subsequent introduction of oligonucleotide targets was accomplished within minutes using a combination of electroosmotic flow and electrophoretic force. The concurrent quantification of femtomole quantities of two target sequences was possible by measuring the spatial coverage of FRET sensitized emission along the length of the channel. In previous reports, multiplexed QD-FRET hybridization assays that employed a ratiometric method for quantification had challenges associated with lower analytical sensitivity arising from both donor and acceptor dilution that resulted in reduced energy transfer pathways as compared to single-color hybridization assays. Herein, a spatial method for quantification that is based on in-channel QD-FRET profiles provided higher analytical sensitivity in the multiplexed assay format as compared to single-color hybridization assays. The selectivity of the multiplexed hybridization assays was demonstrated by discrimination between a fully-complementary sequence and a 3 base pair sequence at a contrast ratio of 8 to 1. Copyright © 2013 Elsevier B.V. All rights reserved.

  1. Paper-based solid-phase nucleic acid hybridization assay using immobilized quantum dots as donors in fluorescence resonance energy transfer.

    Noor, M Omair; Shahmuradyan, Anna; Krull, Ulrich J

    2013-02-05

    A paper-based solid-phase assay is presented for transduction of nucleic acid hybridization using immobilized quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET). The surface of paper was modified with imidazole groups to immobilize QD-probe oligonucleotide conjugates that were assembled in solution. Green-emitting QDs (gQDs) were FRET-paired with Cy3 acceptor. Hybridization of Cy3-labeled oligonucleotide targets provided the proximity required for FRET-sensitized emission from Cy3, which served as an analytical signal. The assay exhibited rapid transduction of nucleic acid hybridization within minutes. Without any amplification steps, the limit of detection of the assay was found to be 300 fmol with the upper limit of the dynamic range at 5 pmol. The implementation of glutathione-coated QDs for the development of nucleic acid hybridization assay integrated on a paper-based platform exhibited excellent resistance to nonspecific adsorption of oligonucleotides and showed no reduction in the performance of the assay in the presence of large quantities of noncomplementary DNA. The selectivity of nucleic acid hybridization was demonstrated by single-nucleotide polymorphism (SNP) detection at a contrast ratio of 19 to 1. The reuse of paper over multiple cycles of hybridization and dehybridization was possible, with less than 20% reduction in the performance of the assay in five cycles. This work provides an important framework for the development of paper-based solid-phase QD-FRET nucleic acid hybridization assays that make use of a ratiometric approach for detection and analysis.

  2. Toward a solid-phase nucleic acid hybridization assay within microfluidic channels using immobilized quantum dots as donors in fluorescence resonance energy transfer.

    Chen, Lu; Algar, W Russ; Tavares, Anthony J; Krull, Ulrich J

    2011-01-01

    The optical properties and surface area of quantum dots (QDs) have made them an attractive platform for the development of nucleic acid biosensors based on fluorescence resonance energy transfer (FRET). Solid-phase assays based on FRET using mixtures of immobilized QD-oligonucleotide conjugates (QD biosensors) have been developed. The typical challenges associated with solid-phase detection strategies include non-specific adsorption, slow kinetics of hybridization, and sample manipulation. The new work herein has considered the immobilization of QD biosensors onto the surfaces of microfluidic channels in order to address these challenges. Microfluidic flow can be used to dynamically control stringency by adjustment of the potential in an electrokinetic-based microfluidics environment. The shearing force, Joule heating, and the competition between electroosmotic and electrophoretic mobilities allow the optimization of hybridization conditions, convective delivery of target to the channel surface to speed hybridization, amelioration of adsorption, and regeneration of the sensing surface. Microfluidic flow can also be used to deliver (for immobilization) and remove QD biosensors. QDs that were conjugated with two different oligonucleotide sequences were used to demonstrate feasibility. One oligonucleotide sequence on the QD was available as a linker for immobilization via hybridization with complementary oligonucleotides located on a glass surface within a microfluidic channel. A second oligonucleotide sequence on the QD served as a probe to transduce hybridization with target nucleic acid in a sample solution. A Cy3 label on the target was excited by FRET using green-emitting CdSe/ZnS QD donors and provided an analytical signal to explore this detection strategy. The immobilized QDs could be removed under denaturing conditions by disrupting the duplex that was used as the surface linker and thus allowed a new layer of QD biosensors to be re-coated within the channel

  3. Paper-based solid-phase multiplexed nucleic acid hybridization assay with tunable dynamic range using immobilized quantum dots as donors in fluorescence resonance energy transfer.

    Noor, M Omair; Krull, Ulrich J

    2013-08-06

    A multiplexed solid-phase nucleic acid hybridization assay on a paper-based platform is presented using multicolor immobilized quantum dots (QDs) as donors in fluorescence resonance energy transfer (FRET). The surface of paper was modified with imidazole groups to immobilize two types of QD-probe oligonucleotide conjugates that were assembled in solution. Green-emitting QDs (gQDs) and red-emitting QDs (rQDs) served as donors with Cy3 and Alexa Fluor 647 (A647) acceptors. The gQD/Cy3 FRET pair served as an internal standard, while the rQD/A647 FRET pair served as a detection channel, combining the control and analytical test zones in one physical location. Hybridization of dye-labeled oligonucleotide targets provided the proximity for FRET sensitized emission from the acceptor dyes, which served as an analytical signal. Hybridization assays in the multicolor format provided a limit of detection of 90 fmol and an upper limit of dynamic range of 3.5 pmol. The use of an array of detection zones was designed to provide improved analytical figures of merit compared to that which could be achieved on one type of array design in terms of relative concentration of multicolor QDs. The hybridization assays showed excellent resistance to nonspecific adsorption of oligonucleotides. Selectivity of the two-plex hybridization assay was demonstrated by single nucleotide polymorphism (SNP) detection at a contrast ratio of 50:1. Additionally, it is shown that the use of preformed QD-probe oligonucleotide conjugates and consideration of the relative number density of the two types of QD-probe conjugates in the two-color assay format is advantageous to maximize assay sensitivity and the upper limit of dynamic range.

  4. Fluorescence resonance energy transfer between NaYF{sub 4}:Yb,Tm upconversion nanoparticles and gold nanorods: Near-infrared responsive biosensor for streptavidin

    Zhang, Shuang; Wang, Jing; Xu, Wen; Chen, Boting; Yu, Wei; Xu, Lin; Song, Hongwei, E-mail: songhw@jlu.edu.cn

    2014-03-15

    We represent a fluorescence resonance energy transfer (FRET) system using upconversion nanoparticles (UCNPs) and the gold nanorods (GNRs) as the energy donor–acceptor pair for directly determining streptavidin in near-infrared (NIR) region. NaYF{sub 4}:Yb,Tm UCNPs, which had a strong emission at 800 nm under 980-nm excitation, were adopted as the energy donor. The GNRs, which demonstrated strong surface plasmon absorption around 800 nm, were chosen as acceptor to quench the 800 nm emissions of the UCNPs. There had the spectral overlap between the emission of the donor nanoparticles (UCNPs) and the absorption of the acceptor nanoparticles (GNRs). This UCNP-based FRET system was then used to determine the amount of streptavidin. In this system, NaYF{sub 4}:Yb,Tm UCNPs conjugated with biotin, while GNRs conjugated with streptavidin. When added GNRs into UCNPs, the streptavidin were preferred to bind with biotin and decreased spacing between the donor and acceptor NPs. Consequently, FRET occurred and a linear relationship between the luminescence quenching efficiency and the concentration of streptavidin was obtained. Owing to the aforementioned merits of UCNPs as an energy donor and the strong quenching ability of GNRs, satisfactory analytical performances have been acquired. -- Highlights: • NaYF4:Yb,Tm and GNRs are as NIR energy donor and quenching acceptor for FRET. • Linkage between biotin and streptavidin make the distance between the donors and the acceptors short enough for FRET. • The FRET system in this work was applicable for the detection of streptavidin. • The donor and acceptor NPs can be modified by proper molecules for other biological molecules detection.

  5. Fluorescence resonance energy transfer between NaYF4:Yb,Tm upconversion nanoparticles and gold nanorods: Near-infrared responsive biosensor for streptavidin

    Zhang, Shuang; Wang, Jing; Xu, Wen; Chen, Boting; Yu, Wei; Xu, Lin; Song, Hongwei

    2014-01-01

    We represent a fluorescence resonance energy transfer (FRET) system using upconversion nanoparticles (UCNPs) and the gold nanorods (GNRs) as the energy donor–acceptor pair for directly determining streptavidin in near-infrared (NIR) region. NaYF 4 :Yb,Tm UCNPs, which had a strong emission at 800 nm under 980-nm excitation, were adopted as the energy donor. The GNRs, which demonstrated strong surface plasmon absorption around 800 nm, were chosen as acceptor to quench the 800 nm emissions of the UCNPs. There had the spectral overlap between the emission of the donor nanoparticles (UCNPs) and the absorption of the acceptor nanoparticles (GNRs). This UCNP-based FRET system was then used to determine the amount of streptavidin. In this system, NaYF 4 :Yb,Tm UCNPs conjugated with biotin, while GNRs conjugated with streptavidin. When added GNRs into UCNPs, the streptavidin were preferred to bind with biotin and decreased spacing between the donor and acceptor NPs. Consequently, FRET occurred and a linear relationship between the luminescence quenching efficiency and the concentration of streptavidin was obtained. Owing to the aforementioned merits of UCNPs as an energy donor and the strong quenching ability of GNRs, satisfactory analytical performances have been acquired. -- Highlights: • NaYF4:Yb,Tm and GNRs are as NIR energy donor and quenching acceptor for FRET. • Linkage between biotin and streptavidin make the distance between the donors and the acceptors short enough for FRET. • The FRET system in this work was applicable for the detection of streptavidin. • The donor and acceptor NPs can be modified by proper molecules for other biological molecules detection

  6. Preliminary Study of the Efficacy of Using Nuclear Resonance Fluorescence with Quasi-Monoenergetic Gamma-Ray Sources for Nuclear Safeguards Assay

    Johnson, M S; McNabb, D P; Hall, J M; Gonzalez, J J

    2011-02-17

    We have studied the efficacy of using nuclear resonance fluorescence (NRF)-based techniques to assay spent nuclear fuel for Pu content using quasi-monoenergetic sources. We have developed two techniques to precisely determine the Pu content in a fuel rod/pin. One of our approaches is virtually free of systematic uncertainties. Using analytical models, we have determined the amount of time required to measure the Pu content in spent nuclear fuel rods and spent fuel assemblies to within 1% precision. We note that Pu content can be determined in a fuel assembly about as fast as in a single fuel pin. The performance of NRF-based assay techniques with improved photon sources, which are currently under development, will also estimated. For follow-on research we propose to: (1) Construct research prototype detection systems for both of the NRF-based assay systems proposed in this paper and measure their calibration curves; (2) Determine the systematic errors associated with both assay methods, explore ways to reduce the errors and fold the results into future performance calculations; (3) Develop an algorithm to assay a fuel assembly; (4) Perform validation measurements using a single pin and scaled assemblies; (5) Research and develop current-mode detection and/or threshold detection techniques to improve assay times; (6) Characterize the flux of newly constructed sources and fold the results into the calculations presented here to determine the feasibility of a variety of proposed sources; and (7) Collaborate with others in the safeguards community to build a prototype system and perform an NRF-based assay demonstration on spent fuel.

  7. Generally Applicable Transformation Protocols for Fluorescent Nanodiamond Internalization into Cells

    Hemelaar, Simon R; van der Laan, Kiran J; Hinterding, Sophie R; Koot, Manon V; Ellermann, Else; Perona-Martinez, Felipe P; Roig, David; Hommelet, Severin; Novarina, Daniele; Takahashi, Hiroki; Chang, Michael; Schirhagl, Romana

    2017-01-01

    Fluorescent nanodiamonds (FNDs) are promising nanoprobes, owing to their stable and magnetosensitive fluorescence. Therefore they can probe properties as magnetic resonances, pressure, temperature or strain. The unprecedented sensitivity of diamond defects can detect the faint magnetic resonance of

  8. From Dark to Light to Fluorescence Resonance Energy Transfer (FRET): Polarity-Sensitive Aggregation-Induced Emission (AIE)-Active Tetraphenylethene-Fused BODIPY Dyes with a Very Large Pseudo-Stokes Shift.

    Şen, Esra; Meral, Kadem; Atılgan, Serdar

    2016-01-11

    The work presented herein is devoted to the fabrication of large Stokes shift dyes in both organic and aqueous media by combining dark resonance energy transfer (DRET) and fluorescence resonance energy transfer (FRET) in one donor-acceptor system. In this respect, a series of donor-acceptor architectures of 4,4-difluoro-4-bora-3a,4a-diaza-s-indacene (BODIPY) dyes substituted by one, two, or three tetraphenylethene (TPE) luminogens were designed and synthesised. The photophysical properties of these three chromophore systems were studied to provide insight into the nature of donor-acceptor interactions in both THF and aqueous media. Because the generation of emissive TPE donor(s) is strongly polarity dependent, due to its aggregation-induced emission (AIE) feature, one might expect the formation of appreciable fluorescence emission intensity with a very large pseudo-Stokes shift in aqueous media when considering FRET process. Interestingly, similar results were also recorded in THF for the chromophore systems, although the TPE fragment(s) of the dyes are non-emissive. The explanation for this photophysical behaviour lies in the DRET. This is the first report on combining two energy-transfer processes, namely, FRET and DRET, in one polarity-sensitive donor-acceptor pair system. The accuracy of the dark-emissive donor property of the TPE luminogen is also presented for the first time as a new feature for AIE phenomena. © 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.

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

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

    2014-01-01

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

  10. A fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) for the detection of mecA gene sequence of Staphylococcus aureus.

    Shi, Jingyu; Chan, Chunyu; Pang, Yukting; Ye, Weiwei; Tian, Feng; Lyu, Jing; Zhang, Yu; Yang, Mo

    2015-05-15

    In this work, a novel fluorescence resonance energy transfer (FRET) biosensor based on graphene quantum dots (GQDs) and gold nanoparticles (AuNPs) pairs was developed for Staphylococcus aureus specific gene sequence detection. This FRET biosensor platform was realized by immobilization of capture probes on GQDs and conjugation of reporter probes on AuNPs. Target oligos then co-hybridized with capture probes and reporter probes to form a sandwich structure which brought GQDs and AuNPs to close proximity to trigger FRET effect. The fluorescence signals before and after addition of targets were measured and the fluorescence quenching efficiency could reach around 87% with 100 nM target oligo. The limit of detection (LOD) of this FRET biosensor was around 1 nM for S.aureus gene detection. Experiments with both single-base mismatched oligos and double-base mismatched oligos demonstrated the good sequence selectivity of this FRET biosensor. Copyright © 2014 Elsevier B.V. All rights reserved.

  11. Correlation between the number of quantum-statistical modes of the exciting field and the number of lines in the resonance fluorescence spectrum

    Kryzhanovskii, Boris V; Sokolov, G B

    2000-01-01

    The quasi-energy wave functions of a two-level atom in an electromagnetic field, the state of which represents a superposition of coherent states, were found. The fluorescence spectrum of an atom excited by such a field was investigated. It was shown that a spectral fluorescence mode corresponds to each mode of the quantum-statistical distribution of the field incident on the atom. This means that the number of statistical modes of the incident field may be recorded as the number of data bits of the information carried by the light pulse. (laser applications and other topics in quantum electronics)

  12. NADH-fluorescence scattering correction for absolute concentration determination in a liquid tissue phantom using a novel multispectral magnetic-resonance-imaging-compatible needle probe

    Braun, Frank; Schalk, Robert; Heintz, Annabell; Feike, Patrick; Firmowski, Sebastian; Beuermann, Thomas; Methner, Frank-Jürgen; Kränzlin, Bettina; Gretz, Norbert; Rädle, Matthias

    2017-07-01

    In this report, a quantitative nicotinamide adenine dinucleotide hydrate (NADH) fluorescence measurement algorithm in a liquid tissue phantom using a fiber-optic needle probe is presented. To determine the absolute concentrations of NADH in this phantom, the fluorescence emission spectra at 465 nm were corrected using diffuse reflectance spectroscopy between 600 nm and 940 nm. The patented autoclavable Nitinol needle probe enables the acquisition of multispectral backscattering measurements of ultraviolet, visible, near-infrared and fluorescence spectra. As a phantom, a suspension of calcium carbonate (Calcilit) and water with physiological NADH concentrations between 0 mmol l-1 and 2.0 mmol l-1 were used to mimic human tissue. The light scattering characteristics were adjusted to match the backscattering attributes of human skin by modifying the concentration of Calcilit. To correct the scattering effects caused by the matrices of the samples, an algorithm based on the backscattered remission spectrum was employed to compensate the influence of multiscattering on the optical pathway through the dispersed phase. The monitored backscattered visible light was used to correct the fluorescence spectra and thereby to determine the true NADH concentrations at unknown Calcilit concentrations. Despite the simplicity of the presented algorithm, the root-mean-square error of prediction (RMSEP) was 0.093 mmol l-1.

  13. Highly-sensitive aptasensor based on fluorescence resonance energy transfer between l-cysteine capped ZnS quantum dots and graphene oxide sheets for the determination of edifenphos fungicide.

    Arvand, Majid; Mirroshandel, Aazam A

    2017-10-15

    With the advantages of excellent optical properties and biocompatibility, single-strand DNA-functionalized quantum dots have been widely applied in biosensing and bioimaging. A new aptasensor with easy operation, high sensitivity, and high selectivity was developed by immobilizing the aptamer on water soluble l-cysteine capped ZnS quantum dots (QDs). Graphene oxide (GO) sheets are mixed with the aptamer-QDs. Consequently, the aptamer-conjugated QDs bind to the GO sheets to form a GO/aptamer-QDs ensemble. This aptasensor enables the energy transfer based on a fluorescence resonance energy transfer (FRET) from the QDs to the GO sheets, quenching the fluorescence of QDs. The GO/aptamer-QDs ensemble assay acts as a "turn-on'' fluorescent sensor for edifenphos (EDI) detection. When GO was replaced by EDI, the fluorescence of QDs was restored and its intensity was proportional to the EDI concentration. This GO-based aptasensor under the optimum conditions exhibited excellent analytical performance for EDI determination, ranging from 5×10 -4 to 6×10 -3 mg L -1 with the detection limit of 1.3×10 -4 mgL -1 . Furthermore, the designed aptasensor exhibited excellent selectivity toward EDI compared to other pesticides and herbicides with similar structures such as diazinon, heptachlor, endrin, dieldrin, butachlor and chlordane. Good reproducibility and precision (RSD =3.9%, n =10) of the assay indicates the high potential of the aptasensor for quantitative trace analysis of EDI. Moreover, the results demonstrate the applicability of the aptasensor for monitoring EDI fungicide in spiked real samples. Copyright © 2017 Elsevier B.V. All rights reserved.

  14. Fluorescence Resonance Energy Transfer of the Tb(III)-Nd(III) Binary System in Molten LiCl-KCl Eutectic Salt

    Kim, B. Y. [Korea Atomic Energy Research Institute, Daejeon (Korea, Republic of); Yun, J. I. [KAIST, Daejeon (Korea, Republic of)

    2015-05-15

    The lanthanides act as a neutron poison in nuclear reactor with large neutron absorption cross section. For that reason, very low amount of lanthanides is required in the recovered U/TRU ingot product from pyrochemical process. In view of that, the investigation of thermodynamic properties and chemical behaviors of lanthanides in molten chloride salt are necessary to estimate the performance efficiency of pyrochemical process. However, there are uncertainties about knowledge and understanding of basic mechanisms in pyrochemical process, such as chemical speciation and redox behaviors due to the lack of in-situ monitoring methods for high temperature molten salt. The spectroscopic analysis is one of the probable techniques for in-situ qualitative and quantitative analysis. Recently, a few fluorescence spectroscopic measurements on single lanthanide element in molten LiCl-KCl eutectic have been investigated. The fluorescence intensity and the fluorescence lifetime of Tb(III) were decreased as increasing the concentration of Nd(III), demonstrating collisional quenching between donor ions and acceptor ions. The Forster distance (..0) of Tb(III)-Nd(III) binary system in molten LiCl-KCl eutectic was determined in the specific range of .... (0.1-1.0) and .. (1.387-1.496)

  15. Fluorescence spectroscopy

    Bagatolli, Luis

    2016-01-01

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

  16. Detection of Babesia canis vogeli and Hepatozoon canis in canine blood by a single-tube real-time fluorescence resonance energy transfer polymerase chain reaction assay and melting curve analysis.

    Kongklieng, Amornmas; Intapan, Pewpan M; Boonmars, Thidarut; Thanchomnang, Tongjit; Janwan, Penchom; Sanpool, Oranuch; Lulitanond, Viraphong; Taweethavonsawat, Piyanan; Chungpivat, Sudchit; Maleewong, Wanchai

    2015-03-01

    A real-time fluorescence resonance energy transfer polymerase chain reaction (qFRET PCR) coupled with melting curve analysis was developed for detection of Babesia canis vogeli and Hepatozoon canis infections in canine blood samples in a single tube assay. The target of the assay was a region within the 18S ribosomal RNA gene amplified in either species by a single pair of primers. Following amplification from the DNA of infected dog blood, a fluorescence melting curve analysis was done. The 2 species, B. canis vogeli and H. canis, could be detected and differentiated in infected dog blood samples (n = 37) with high sensitivity (100%). The detection limit for B. canis vogeli was 15 copies of a positive control plasmid, and for H. canis, it was 150 copies of a positive control plasmid. The assay could simultaneously distinguish the DNA of both parasites from the DNA of controls. Blood samples from 5 noninfected dogs were negative, indicating high specificity. Several samples can be run at the same time. The assay can reduce misdiagnosis and the time associated with microscopic examination, and is not prone to the carryover contamination associated with the agarose gel electrophoresis step of conventional PCR. In addition, this qFRET PCR method would be useful to accurately determine the range of endemic areas or to discover those areas where the 2 parasites co-circulate. © 2015 The Author(s).

  17. Conformation of L-Tyrosine Studied by Fluorescence-Detected UV-UV and IR-UV Double-Resonance Spectroscopy

    Inokuchi, Yoshiya; Kobayashi, Yusuke; Ito, Takafumi; Ebata, Takayuki

    2007-01-01

    The laser-induced fluorescence spectrum of jet-cooled L-tyrosine exhibits more than 20 vibronic bands in the 35450-35750 cm-1 region. We attribute these bands to eight conformers by using results of UV-UV hole-burning spectroscopy. These isomers are classified into four groups; each group consists of two rotational isomers that have a similar side-chain conformation but different orientations of the phenolic OH. The splitting of band origins of rotational isomers is 31, 21, 5, and 0 cm-1 for ...

  18. Fluorescence fluctuation spectroscopy (FFS)

    Tetin, Sergey

    2012-01-01

    This new volume of Methods in Enzymology continues the legacy of this premier serial with quality chapters authored by leaders in the field. This volume covers fluorescence fluctuation spectroscopy and includes chapters on such topics as Förster resonance energy transfer (fret) with fluctuation algorithms, protein corona on nanoparticles by FCS, and FFS approaches to the study of receptors in live cells. Continues the legacy of this premier serial with quality chapters authored by leaders in the field Covers fluorescence fluctuation spectroscopy Contains chapters on such topics as Förster resonance energy transfer (fret) with fluctuation algorithms, protein corona on nanoparticles by FCS, and FFS approaches to the study of receptors in live cells.

  19. Charge recombination process in X-ray irradiated pyrene-doped polystyrene as studied by optically detected electron spin resonance and magnetic field dependence of the recombination fluorescence

    Okazaki, Masaharu; Tai, Yutaka; Toriyama, Kazumi

    1993-01-01

    The optically-detected ESR (ODESR) spectrum and magnetic field dependence on recombination fluorescence were observed for X-ray irradiated pyrene-doped polystyrene at temperatures of 242-348 K. The ODESR intensity as a function of the pyrene concentration, 0.1-8.9 wt%, showed an unusual minimum at about 1.0%. Two phases were separated in the magnetic field dependence of the fluorescence: one was sharp and saturates at fields of over 50 mT, while the other was broad with a dip at around 60-150 mT. The cause of this dip was naturally attributed to the ST -1 level crossing. The sharp magnetic field effect also showed a minimum at around a concentration of 1.0 wt%. These novel findings have been interpreted using a recombination model modified from the previous one for pyrene-doped ethylene-propylene rubber and polyethylene. The essential points of the present model are: (1) although electron hopping within the polystyrene molecule is rapid, electron transfer at the last step of recombination between the polystyrene anion and the pyrene cation proceeds at a moderate rate; (2) the hole-transfer rate in the polymer chain is moderate; (3) electron hopping between the doped pyrene molecules is very much dependent on the concentration; (4) hole hopping between the pyrenes is inhibited. (author)

  20. "Smart" theranostic lanthanide nanoprobes with simultaneous up-conversion fluorescence and tunable T1-T2 magnetic resonance imaging contrast and near-infrared activated photodynamic therapy.

    Zhang, Yan; Das, Gautom Kumar; Vijayaragavan, Vimalan; Xu, Qing Chi; Padmanabhan, Parasuraman; Bhakoo, Kishore K; Selvan, Subramanian Tamil; Tan, Timothy Thatt Yang

    2014-11-07

    The current work reports a type of "smart" lanthanide-based theranostic nanoprobe, NaDyF4:Yb(3+)/NaGdF4:Yb(3+),Er(3+), which is able to circumvent the up-converting poisoning effect of Dy(3+) ions to give efficient near infrared (980 nm) triggered up-conversion fluorescence, and offers not only excellent dark T2-weighted MR contrast but also tunable bright and T1-weighted MR contrast properties. Due to the efficient up-converted energy transfer from the nanocrystals to chlorin e6 (Ce6) photosensitizers loaded onto the nanocrystals, cytotoxic singlet oxygen was generated and photodynamic therapy was demonstrated. Therefore, the current multifunctional nanocrystals could be potentially useful in various image-guided diagnoses where bright or dark MRI contrast could be selectively tuned to optimize image quality, but also as an efficient and more penetrative near-infrared activated photodynamic therapy agent.

  1. ``Smart'' theranostic lanthanide nanoprobes with simultaneous up-conversion fluorescence and tunable T1-T2 magnetic resonance imaging contrast and near-infrared activated photodynamic therapy

    Zhang, Yan; Das, Gautom Kumar; Vijayaragavan, Vimalan; Xu, Qing Chi; Padmanabhan, Parasuraman; Bhakoo, Kishore K.; Tamil Selvan, Subramanian; Tan, Timothy Thatt Yang

    2014-10-01

    The current work reports a type of ``smart'' lanthanide-based theranostic nanoprobe, NaDyF4:Yb3+/NaGdF4:Yb3+,Er3+, which is able to circumvent the up-converting poisoning effect of Dy3+ ions to give efficient near infrared (980 nm) triggered up-conversion fluorescence, and offers not only excellent dark T2-weighted MR contrast but also tunable bright and T1-weighted MR contrast properties. Due to the efficient up-converted energy transfer from the nanocrystals to chlorin e6 (Ce6) photosensitizers loaded onto the nanocrystals, cytotoxic singlet oxygen was generated and photodynamic therapy was demonstrated. Therefore, the current multifunctional nanocrystals could be potentially useful in various image-guided diagnoses where bright or dark MRI contrast could be selectively tuned to optimize image quality, but also as an efficient and more penetrative near-infrared activated photodynamic therapy agent.The current work reports a type of ``smart'' lanthanide-based theranostic nanoprobe, NaDyF4:Yb3+/NaGdF4:Yb3+,Er3+, which is able to circumvent the up-converting poisoning effect of Dy3+ ions to give efficient near infrared (980 nm) triggered up-conversion fluorescence, and offers not only excellent dark T2-weighted MR contrast but also tunable bright and T1-weighted MR contrast properties. Due to the efficient up-converted energy transfer from the nanocrystals to chlorin e6 (Ce6) photosensitizers loaded onto the nanocrystals, cytotoxic singlet oxygen was generated and photodynamic therapy was demonstrated. Therefore, the current multifunctional nanocrystals could be potentially useful in various image-guided diagnoses where bright or dark MRI contrast could be selectively tuned to optimize image quality, but also as an efficient and more penetrative near-infrared activated photodynamic therapy agent. Electronic supplementary information (ESI) available. See DOI: 10.1039/c4nr01717j

  2. Fluorescence resonance energy transfer measured by spatial photon migration in CdSe-ZnS quantum dots colloidal systems as a function of concentration

    Azevedo, G.; Monte, A. F. G.; Reis, A. F.; Messias, D. N. [Laboratório de Espectroscopia Óptica, Instituto de Física, Universidade Federal de Uberlândia, Uberlândia, MG 38400-902 (Brazil)

    2014-11-17

    The study of the spatial photon migration as a function of the concentration brings into attention the problem of the energy transfer in quantum dot embedded systems. By measuring the photon propagation and its spatial dependence, it is possible to understand the whole dynamics in a quantum dot system, and also improve their concentration dependence to maximize energy propagation due to radiative and non-radiative processes. In this work, a confocal microscope was adapted to scan the spatial distribution of photoluminescence from CdSe-ZnS core-shell quantum dots in colloidal solutions. The energy migration between the quantum dots was monitored by the direct measurement of the photon diffusion length, according to the diffusion theory. We observed that the photon migration length decreases by increasing the quantum dot concentration, this kind of behavior has been regarded as a signature of Förster resonance energy transfer in the system.

  3. Fourier Transform Near Infrared Microspectroscopy, Infrared Chemical Imaging, High-Resolution Nuclear Magnetic Resonance and Fluorescence Microspectroscopy Detection of Single Cancer Cells and Single Viral Particles

    Baianu,I C; Hofmann, N E; Korban, S S; Lozano, P; You, T

    2004-01-01

    Single Cancer Cells from Human tumors are being detected and imaged by Fourier Transform Infrared (FT-IR), Fourier Transform Near Infrared (FT-NIR)Hyperspectral Imaging and Fluorescence Correlation Microspectroscopy. The first FT-NIR chemical, microscopic images of biological systems approaching one micron resolution are here reported. Chemical images obtained by FT-NIR and FT-IR Microspectroscopy are also presented for oil in soybean seeds and somatic embryos under physiological conditions. FT-NIR spectra of oil and proteins were obtained for volumes as small as two cubic microns. Related, HR-NMR analyses of oil contents in somatic embryos as well as 99% accurate calibrations are also presented here with nanoliter precision. Such high-resolution, 400 MHz H-1 NMR analyses allowed the selection of mutagenized embryos with higher oil content (e.g. >~20%) compared to the average levels in non-mutagenized control embryos. Moreover, developmental changes in single soybean seeds and/or somatic embryos may be monito...

  4. Near Infrared Microspectroscopy, Fluorescence Microspectroscopy, Infrared Chemical Imaging and High Resolution Nuclear Magnetic Resonance Analysis of Soybean Seeds, Somatic Embryos and Single Cells

    Baianu, I C; Hofmann, N E; Korban, S S; Lozano, P; You, T; AOCS 94th Meeting, Kansas

    2002-01-01

    Novel methodologies are currently being developed and established for the chemical analysis of soybean seeds, embryos and single cells by Fourier Transform Infrared (FT-IR), Fourier Transform Near Infrared (FT-NIR) Microspectroscopy, Fluorescence and High-Resolution NMR (HR-NMR). The first FT-NIR chemical images of biological systems approaching one micron resolution are presented here. Chemical images obtained by FT-NIR and FT-IR Microspectroscopy are presented for oil in soybean seeds and somatic embryos under physiological conditions. FT-NIR spectra of oil and proteins were obtained for volumes as small as two cubic microns. Related, HR-NMR analyses of oil contents in somatic embryos are also presented here with nanoliter precision. Such 400 MHz 1H NMR analyses allowed the selection of mutagenized embryos with higher oil content (e.g. ~20%) compared to non-mutagenized control embryos. Moreover, developmental changes in single soybean seeds and/or somatic embryos may be monitored by FT-NIR with a precision ...

  5. Fluorescence resonance energy transfer-based real-time polymerase chain reaction method without DNA extraction for the genotyping of F5, F2, F12, MTHFR, and HFE.

    Martinez-Serra, Jordi; Robles, Juan; Nicolàs, Antoni; Gutierrez, Antonio; Ros, Teresa; Amat, Juan Carlos; Alemany, Regina; Vögler, Oliver; Abelló, Aina; Noguera, Aina; Besalduch, Joan

    2014-01-01

    Blood samples are extensively used for the molecular diagnosis of many hematological diseases. The daily practice in a clinical laboratory of molecular diagnosis in hematology involves using a variety of techniques, based on the amplification of nucleic acids. Current methods for polymerase chain reaction (PCR) use purified genomic DNA, mostly isolated from total peripheral blood cells or white blood cells (WBC). In this paper we describe a real-time fluorescence resonance energy transfer-based method for genotyping directly from blood cells. Our strategy is based on an initial isolation of the WBCs, allowing the removal of PCR inhibitors, such as the heme group, present in the erythrocytes. Once the erythrocytes have been lysed, in the LightCycler(®) 2.0 Instrument, we perform a real-time PCR followed by a melting curve analysis for different genes (Factors 2, 5, 12, MTHFR, and HFE). After testing 34 samples comparing the real-time crossing point (CP) values between WBC (5×10(6) WBC/mL) and purified DNA (20 ng/μL), the results for F5 Leiden were as follows: CP mean value for WBC was 29.26±0.566 versus purified DNA 24.79±0.56. Thus, when PCR was performed from WBC (5×10(6) WBC/mL) instead of DNA (20 ng/μL), we observed a delay of about 4 cycles. These small differences in CP values were similar for all genes tested and did not significantly affect the subsequent analysis by melting curves. In both cases the fluorescence values were high enough, allowing a robust genotyping of all these genes without a previous DNA purification/extraction.

  6. A singleplex real-time fluorescence resonance energy transfer PCR with melting curve analysis for the differential detection of Paragonimus heterotremus, Echinostoma malayanum and Fasciola gigantica eggs in faeces.

    Tantrawatpan, Chairat; Saijuntha, Weerachai; Manochantr, Sirikul; Kheolamai, Pakpoom; Thanchomnang, Tongjit; Sadaow, Lakkhana; Intapan, Pewpan M; Maleewong, Wanchai

    2016-01-01

    Because the eggs of Paragonimus, Echinostoma and Fasciola are very similar in size and shape, it is difficult to distinguish and accurately identify species by the morphology of their eggs, which is a standard diagnostic method. In this study, a novel assay combining a real-time fluorescence resonance energy transfer PCR and melting curve analysis using one set of primers and fluorophore-labelled hybridization probes specific for the 28S rDNA region was developed for the molecular detection of Paragonimus heterotremus, Echinostoma malayanum and Fasciola gigantica eggs. This assay could detect and distinguish P. heterotremus, E. malayanum and F. gigantica DNA with the distinct melting temperature (Tm) values of 57.99±0.08, 62.12±0.15 and 74.10±0.18, respectively. The assay can also be used to detect and distinguish DNA from P. bangkokensis, P. harinasutai, P. machorchis, E. revolutum, Hypodereum conoideum and F. hepatica, which have different Tm values. The sensitivity of this assay enabled the detection of one egg of P. heterotremus, E. malayanum or F. gigantica per 100 mg of faeces. In addition, the specificity testing showed no fluorescence signal for other parasites. Due to the sensitivity and specificity of our assay in detecting P. heterotremus, E. malayanum and F. gigantica, our method could be used to accurately diagnose these three medically important parasitic groups and has potential implications for molecular epidemiological investigations of human and/or animal infections. © The Author 2015. Published by Oxford University Press on behalf of Royal Society of Tropical Medicine and Hygiene. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.

  7. Comparison of two DNA targets for the diagnosis of Toxoplasmosis by real-time PCR using fluorescence resonance energy transfer hybridization probes

    Ernault Pauline

    2003-05-01

    Full Text Available Abstract Background Toxoplasmosis is an infectious disease caused by the parasitic protozoan Toxoplasma gondii. It is endemic worldwide and, depending on the geographic location, 15 to 85% of the human population are asymptomatically infected. Routine diagnosis is based on serology. The parasite has emerged as a major opportunistic pathogen for immunocompromised patients, in whom it can cause life-threatening disease. Moreover, when a pregnant woman develops a primary Toxoplasma gondii infection, the parasite may be transmitted to the fetus and cause serious damnage. For these two subpopulations, a rapid and accurate diagnosis is required to initiate treatment. Serological diagnosis of active infection is unreliable because reactivation is not always accompanied by changes in antibody levels, and the presence of IgM does not necessarily indicate recent infection. Application of quantitative PCR has evolved as a sensitive, specific, and rapid method for the detection of Toxoplasma gondii DNA in amniotic fluid, blood, tissue samples, and cerebrospinal fluid. Methods Two separate, real-time fluorescence PCR assays were designed and evaluated with clinical samples. The first, targeting the 35-fold repeated B1 gene, and a second, targeting a newly described multicopy genomic fragment of Toxoplasma gondii. Amplicons of different intragenic copies were analyzed for sequence heterogeneity. Results Comparative LightCycler experiments were conducted with a dilution series of Toxoplasma gondii genomic DNA, 5 reference strains, and 51 Toxoplasma gondii-positive amniotic fluid samples revealing a 10 to 100-fold higher sensitivity for the PCR assay targeting the newly described 529-bp repeat element of Toxoplasma gondii. Conclusion We have developed a quantitative LightCycler PCR protocol which offer rapid cycling with real-time, sequence-specific detection of amplicons. Results of quantitative PCR demonstrate that the 529-bp repeat element is repeated more

  8. Modulation of Intracellular Quantum Dot to Fluorescent Protein Förster Resonance Energy Transfer via Customized Ligands and Spatial Control of Donor–Acceptor Assembly

    Lauren D. Field

    2015-12-01

    Full Text Available Understanding how to controllably modulate the efficiency of energy transfer in Förster resonance energy transfer (FRET-based assemblies is critical to their implementation as sensing modalities. This is particularly true for sensing assemblies that are to be used as the basis for real time intracellular sensing of intracellular processes and events. We use a quantum dot (QD donor -mCherry acceptor platform that is engineered to self-assemble in situ wherein the protein acceptor is expressed via transient transfection and the QD donor is microinjected into the cell. QD-protein assembly is driven by metal-affinity interactions where a terminal polyhistidine tag on the protein binds to the QD surface. Using this system, we show the ability to modulate the efficiency of the donor–acceptor energy transfer process by controllably altering either the ligand coating on the QD surface or the precise location where the QD-protein assembly process occurs. Intracellularly, a short, zwitterionic ligand mediates more efficient FRET relative to longer ligand species that are based on the solubilizing polymer, poly(ethylene glycol. We further show that a greater FRET efficiency is achieved when the QD-protein assembly occurs free in the cytosol compared to when the mCherry acceptor is expressed tethered to the inner leaflet of the plasma membrane. In the latter case, the lower FRET efficiency is likely attributable to a lower expression level of the mCherry acceptor at the membrane combined with steric hindrance. Our work points to some of the design considerations that one must be mindful of when developing FRET-based sensing schemes for use in intracellular sensing.

  9. Dual-Recognition Förster Resonance Energy Transfer Based Platform for One-Step Sensitive Detection of Pathogenic Bacteria Using Fluorescent Vancomycin-Gold Nanoclusters and Aptamer-Gold Nanoparticles.

    Yu, Mengqun; Wang, Hong; Fu, Fei; Li, Linyao; Li, Jing; Li, Gan; Song, Yang; Swihart, Mark T; Song, Erqun

    2017-04-04

    The effective monitoring, identification, and quantification of pathogenic bacteria is essential for addressing serious public health issues. In this study, we present a universal and facile one-step strategy for sensitive and selective detection of pathogenic bacteria using a dual-molecular affinity-based Förster (fluorescence) resonance energy transfer (FRET) platform based on the recognition of bacterial cell walls by antibiotic and aptamer molecules, respectively. As a proof of concept, Vancomycin (Van) and a nucleic acid aptamer were employed in a model dual-recognition scheme for detecting Staphylococcus aureus (Staph. aureus). Within 30 min, by using Van-functionalized gold nanoclusters and aptamer-modified gold nanoparticles as the energy donor and acceptor, respectively, the FRET signal shows a linear variation with the concentration of Staph. aureus in the range from 20 to 10 8 cfu/mL with a detection limit of 10 cfu/mL. Other nontarget bacteria showed negative results, demonstrating the good specificity of the approach. When employed to assay Staph. aureus in real samples, the dual-recognition FRET strategy showed recoveries from 99.00% to the 109.75% with relative standard derivations (RSDs) less than 4%. This establishes a universal detection platform for sensitive, specific, and simple pathogenic bacteria detection, which could have great impact in the fields of food/public safety monitoring and infectious disease diagnosis.

  10. Fluorogenic Cell-Based Biosensors for Monitoring Microbes

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

    2010-01-01

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

  11. Fluorescence resonance energy transfer-based real-time polymerase chain reaction method without DNA extraction for the genotyping of F5, F2, F12, MTHFR, and HFE

    Martinez-Serra J

    2014-06-01

    Full Text Available Jordi Martinez-Serra,1 Juan Robles,2 Antoni Nicolàs,3 Antonio Gutierrez,1 Teresa Ros,1 Juan Carlos Amat,1 Regina Alemany,4 Oliver Vögler,4 Aina Abelló,2 Aina Noguera,2 Joan Besalduch1 1Department of Hematology, 2Department of Clinical Analysis, Hospital Universitary Son Espases, Palma de Mallorca, Spain; 3ECOGEN, Barcelona, 4Department of Cell Biology, University of the Balearic Islands, Palma de Mallorca, Spain Abstract: Blood samples are extensively used for the molecular diagnosis of many hematological diseases. The daily practice in a clinical laboratory of molecular diagnosis in hematology involves using a variety of techniques, based on the amplification of nucleic acids. Current methods for polymerase chain reaction (PCR use purified genomic DNA, mostly isolated from total peripheral blood cells or white blood cells (WBC. In this paper we describe a real-time fluorescence resonance energy transfer-based method for genotyping directly from blood cells. Our strategy is based on an initial isolation of the WBCs, allowing the removal of PCR inhibitors, such as the heme group, present in the erythrocytes. Once the erythrocytes have been lysed, in the LightCycler® 2.0 Instrument, we perform a real-time PCR followed by a melting curve analysis for different genes (Factors 2, 5, 12, MTHFR, and HFE. After testing 34 samples comparing the real-time crossing point (CP values between WBC (5×106 WBC/mL and purified DNA (20 ng/µL, the results for F5 Leiden were as follows: CP mean value for WBC was 29.26±0.566 versus purified DNA 24.79±0.56. Thus, when PCR was performed from WBC (5×106 WBC/mL instead of DNA (20 ng/µL, we observed a delay of about 4 cycles. These small differences in CP values were similar for all genes tested and did not significantly affect the subsequent analysis by melting curves. In both cases the fluorescence values were high enough, allowing a robust genotyping of all these genes without a previous DNA purification

  12. Fluorescence microscopy.

    Sanderson, Michael J; Smith, Ian; Parker, Ian; Bootman, Martin D

    2014-10-01

    Fluorescence microscopy is a major tool with which to monitor cell physiology. Although the concepts of fluorescence and its optical separation using filters remain similar, microscope design varies with the aim of increasing image contrast and spatial resolution. The basics of wide-field microscopy are outlined to emphasize the selection, advantages, and correct use of laser scanning confocal microscopy, two-photon microscopy, scanning disk confocal microscopy, total internal reflection, and super-resolution microscopy. In addition, the principles of how these microscopes form images are reviewed to appreciate their capabilities, limitations, and constraints for operation. © 2014 Cold Spring Harbor Laboratory Press.

  13. Water simulation for cell based sandbox games

    Lundell, Christian

    2014-01-01

    This thesis work presents a new algorithm for simulating fluid based on the Navier-Stokes equations. The algorithm is designed for cell based sandbox games where interactivity and performance are the main priorities. The algorithm enforces mass conservation conservatively instead of enforcing a divergence free velocity field. A global scale pressure model that simulates hydrostatic pressure is used where the pressure propagates between neighboring cells. A prefix sum algorithm is used to only...

  14. Ontology for cell-based geographic information

    Zheng, Bin; Huang, Lina; Lu, Xinhai

    2009-10-01

    Inter-operability is a key notion in geographic information science (GIS) for the sharing of geographic information (GI). That requires a seamless translation among different information sources. Ontology is enrolled in GI discovery to settle the semantic conflicts for its natural language appearance and logical hierarchy structure, which are considered to be able to provide better context for both human understanding and machine cognition in describing the location and relationships in the geographic world. However, for the current, most studies on field ontology are deduced from philosophical theme and not applicable for the raster expression in GIS-which is a kind of field-like phenomenon but does not physically coincide to the general concept of philosophical field (mostly comes from the physics concepts). That's why we specifically discuss the cell-based GI ontology in this paper. The discussion starts at the investigation of the physical characteristics of cell-based raster GI. Then, a unified cell-based GI ontology framework for the recognition of the raster objects is introduced, from which a conceptual interface for the connection of the human epistemology and the computer world so called "endurant-occurrant window" is developed for the better raster GI discovery and sharing.

  15. Nanoantenna array-induced fluorescence enhancement and reduced lifetimes

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

    2008-01-01

    Enhanced fluorescence is observed from dye molecules interacting with optical nanoantenna arrays. Elliptical gold dimers form individual nanoantennae with tunable plasmon resonances depending upon the geometry of the two particles and the size of the gap between them. A fluorescent dye, Rhodamine...... 800, is uniformly embedded in a dielectric host that coats the nanoantennae. The nanoantennae act to enhance the dye absorption. In turn, emission from the dye drives the plasmon resonance of the antennae; the nanoantennae act to enhance the fluorescence signal and change the angular distribution...... of emission. These effects depend upon the overlap of the plasmon resonance with the excitation wavelength and the fluorescence emission band. A decreased fluorescence lifetime is observed along with highly polarized emission that displays the characteristics of the nanoantenna's dipole mode. Being able...

  16. Polarization of fluorescence: a probe of molecular autoionization

    Leroi, G.E.; Dehmer, J.L.; Parr, A.C.; Poliakoff, E.D.

    1983-01-01

    The polarization of fluorescence from excited-state molecular photoions provides a direct probe of the photoionization dynamics and the symmetry signatures of autoionizing resonances. Measurements on CO 2 and CS 2 are presented as examples

  17. Stem cell-based approaches in dentistry

    TA Mitsiadis

    2011-11-01

    Full Text Available Repair of dental pulp and periodontal lesions remains a major clinical challenge. Classical dental treatments require the use of specialised tissue-adapted materials with still questionable efficacy and durability. Stem cell-based therapeutic approaches could offer an attractive alternative in dentistry since they can promise physiologically improved structural and functional outcomes. These therapies necessitate a sufficient number of specific stem cell populations for implantation. Dental mesenchymal stem cells can be easily isolated and are amenable to in vitro expansion while retaining their stemness. In vivo studies realised in small and large animals have evidenced the potential of dental mesenchymal stem cells to promote pulp and periodontal regeneration, but have also underlined new important challenges. The homogeneity of stem cell populations and their quality control, the delivery method, the quality of the regenerated dental tissues and their integration to the host tissue are some of the key challenges. The use of bioactive scaffolds that can elicit effective tissue repair response, through activation and mobilisation of endogenous stem cell populations, constitutes another emerging therapeutic strategy. Finally, the use of stem cells and induced pluripotent cells for the regeneration of entire teeth represents a novel promising alternative to dental implant treatment after tooth loss. In this mini-review, we present the currently applied techniques in restorative dentistry and the various attempts that are made to bridge gaps in knowledge regarding treatment strategies by translating basic stem cell research into the dental practice.

  18. Random search for a dark resonance

    Kiilerich, Alexander Holm; Mølmer, Klaus

    A pair of resonant laser fields can drive a three-level system into a dark state where it ceases to absorb and emit radiation due to destructive interference. We propose a scheme to search for this resonance by randomly changing the frequency of one of the fields each time a fluorescence photon...

  19. Cell-based fluorescence assay for human immunodeficiency virus type 1 protease activity

    Lindsten, K.; Kondrová, Taťána; Konvalinka, Jan; Masucci, M. G.; Dantuma, N. P.

    2001-01-01

    Roč. 45, č. 9 (2001), s. 2616-2622 ISSN 0066-4804 R&D Projects: GA ČR GA303/98/1559 Grant - others:HHMI(US) 75195-540801; ECTM(XE) ERBFMRXCT960026 Institutional research plan: CEZ:AV0Z4055905 Keywords : HIV-1 * protease activity Subject RIV: CE - Biochemistry Impact factor: 4.562, year: 2001

  20. Multiphoton resonances

    Shore, B.W.

    1977-01-01

    The long-time average of level populations in a coherently-excited anharmonic sequence of energy levels (e.g., an anharmonic oscillator) exhibits sharp resonances as a function of laser frequency. For simple linearly-increasing anharmonicity, each resonance is a superposition of various multiphoton resonances (e.g., a superposition of 3, 5, 7, . . . photon resonances), each having its own characteristic width predictable from perturbation theory

  1. Progenitor cell-based treatment of glial disease

    Goldman, Steven A

    2017-01-01

    -based neurodegenerative conditions may now be compelling targets for cell-based therapy. As such, glial cell-based therapies may offer potential benefit to a broader range of diseases than ever before contemplated, including disorders such as Huntington's disease and the motor neuron degeneration of amyotrophic lateral...

  2. Spectroscopy and nonclassical fluorescence properties of single trapped Ba+ ions

    Bolle, J.

    1998-06-01

    This thesis reports on the setup and application of an experimental apparatus for spectroscopic and quantum optical investigations of a single Barium ion in a Paul trap. The realization of the apparatus, which consists of the ion trap in ultra high vacuum, two laser systems, and a photon counting detection system, is described in detail, with particular consideration of the noise sources like stray light and laser frequency instabilities. The two lasers at 493 nm and 650 nm needed to continuously excite resonance fluorescence from the Barium ion have been realized using diode lasers only. The preparation of a single localized Barium ion is described, in particular its optical cooling with the laser light and the minimization of induced vibration in the trapping potential. The purely quantum mechanical property of antibunching is observed by measuring the intensity correlation function of resonance fluorescence from the trapped and cooled ion. Interference properties of the single ion resonance fluorescence are investigated with a Mach-Zehnder interferometer. From the measured high-contrast interference signal it is proven that each individual fluorescence photon interferes with itself. The fluorescence excitation spectrum, on varying one laser frequency, is also measured and exhibits dark resonances. These measurements are compared to calculations based on optical Bloch equations for the 8 atomic levels involved. Future experiments, in particular the detection of reduced quantum fluctuations (squeezing) in one quadrature component of the resonance fluorescence, are discussed. (author)

  3. Reviews in fluorescence 2010

    Geddes, Chris D

    2011-01-01

    ""Reviews in Fluorescence 2010"", the seventh volume of the book serial from Springer, serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence and closely related disciplines. It summarizes the year's progress in fluorescence and its applications, with authoritative analytical reviews specialized enough to be attractive to professional researchers, yet also appealing to the wider audience of scientists in related disciplines of fluorescence. ""Reviews in Fluorescence"" offers an essential reference material for any lab working in the fluoresc

  4. Principles of fluorescence techniques

    2016-01-01

    Fluorescence techniques are being used and applied increasingly in academics and industry. The Principles of Fluorescence Techniques course will outline the basic concepts of fluorescence techniques and the successful utilization of the currently available commercial instrumentation. The course is designed for students who utilize fluorescence techniques and instrumentation and for researchers and industrial scientists who wish to deepen their knowledge of fluorescence applications. Key scientists in the field will deliver theoretical lectures. The lectures will be complemented by the direct utilization of steady-state and lifetime fluorescence instrumentation and confocal microscopy for FLIM and FRET applications provided by leading companies.

  5. Plasmonic enhancement of ultraviolet fluorescence

    Jiao, Xiaojin

    Plasmonics relates to the interaction between electromagnetic radiation and conduction electrons at metallic interfaces or in metallic nanostructures. Surface plasmons are collective electron oscillations at a metal surface, which can be manipulated by shape, texture and material composition. Plasmonic applications cover a broad spectrum from visible to near infrared, including biosensing, nanolithography, spectroscopy, optoelectronics, photovoltaics and so on. However, there remains a gap in this activity in the ultraviolet (UV, research. Motivating factors in the study of UV Plasmonics are the direct access to biomolecular resonances and native fluorescence, resonant Raman scattering interactions, and the potential for exerting control over photochemical reactions. This dissertation aims to fill in the gap of Plasmonics in the UV with efforts of design, fabrication and characterization of aluminium (Al) and magnesium (Mg) nanostructures for the application of label-free bimolecular detection via native UV fluorescence. The first contribution of this dissertation addresses the design of Al nanostructures in the context of UV fluorescence enhancement. A design method that combines analytical analysis with numerical simulation has been developed. Performance of three canonical plasmonic structures---the dipole antenna, bullseye nanoaperture and nanoaperture array---has been compared. The optimal geometrical parameters have been determined. A novel design of a compound bullseye structure has been proposed and numerically analyzed for the purpose of compensating for the large Stokes shift typical of UV fluorescence. Second, UV lifetime modification of diffusing molecules by Al nanoapertures has been experimentally demonstrated for the first time. Lifetime reductions of ~3.5x have been observed for the high quantum yield (QY) laser dye p-terphenyl in a 60 nm diameter aperture with 50 nm undercut. Furthermore, quantum-yield-dependence of lifetime reduction has been

  6. Dipole Resonances of 76Ge

    Ilieva, R. S.; Cooper, N.; Werner, V.; Rusev, G.; Pietralla, N.; Kelly, J. H.; Tornow, W.; Yates, S. W.; Crider, B. P.; Peters, E.

    2013-10-01

    Dipole resonances in 76Ge have been studied using the method of Nuclear Resonance Fluorescence (NRF). The experiment was performed using the Free Electron Laser facility at HI γS/TUNL, which produced linearly polarised quasi-monoenergetic photons in the 4-9 MeV energy range. Photon strength, in particular dipole strength, is an important ingredient in nuclear reaction calculations, and recent interest in its study has been stimulated by observations of a pygmy dipole resonance near the neutron separation energy Sn of certain nuclei. Furthermore, 76Ge is a candidate for 0 ν 2 β -decay. The results are complimentary to a relevant experiment done at TU Darmstadt using Bremsstrahlung beams. Single-resonance parities and a preliminary estimate of the total photo-excitation cross section will be presented. This work was supported by the U.S. DOE under grant no. DE-FG02-91ER40609.

  7. Reviews in fluorescence 2008

    Geddes, Chris D

    2010-01-01

    This volume serves as a comprehensive collection of current trends and emerging hot topics in the field of fluorescence spectroscopy. It summarizes the year's progress in fluorescence and its applications as well as includes authoritative analytical reviews.

  8. Fluorescent optical position sensor

    Weiss, Jonathan D.

    2005-11-15

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

  9. a new analytical modeling method for photovoltaic solar cells based

    Zieba Falama R, Dadjé A, Djongyang N and Doka S.Y

    CELLS BASED ON DERIVATIVE POWER FUNCTION ... Received: 25 Jaunary 2016 / Accepted: 25 April 2016 / Published online: 01 May 2016 ..... For the simulation, two electric PV modules for different technologies were used; these.

  10. Synchrobetatron resonances

    1977-03-01

    At the 1975 Particle Accelerator Conference it was reported that a class of resonances were observed in SPEAR II that had not appeared before in SPEAR I. While the existence of sideband resonances of the main betatron oscillation frequencies has been previously observed and analyzed, the resonances observed in SPEAR do not appear to be of the same variety. Experiments were performed at SPEAR to identify the mechanism believed to be the most likely explanation. Some of the current experimental knowledge and theoretical views on the source of these resonances are presented

  11. Snake resonances

    Tepikian, S.

    1988-01-01

    Siberian Snakes provide a practical means of obtaining polarized proton beams in large accelerators. The effect of snakes can be understood by studying the dynamics of spin precession in an accelerator with snakes and a single spin resonance. This leads to a new class of energy independent spin depolarizing resonances, called snake resonances. In designing a large accelerator with snakes to preserve the spin polarization, there is an added constraint on the choice of the vertical betatron tune due to the snake resonances. 11 refs., 4 figs

  12. Safe biodegradable fluorescent particles

    Martin, Sue I [Berkeley, CA; Fergenson, David P [Alamo, CA; Srivastava, Abneesh [Santa Clara, CA; Bogan, Michael J [Dublin, CA; Riot, Vincent J [Oakland, CA; Frank, Matthias [Oakland, CA

    2010-08-24

    A human-safe fluorescence particle that can be used for fluorescence detection instruments or act as a safe simulant for mimicking the fluorescence properties of microorganisms. The particle comprises a non-biological carrier and natural fluorophores encapsulated in the non-biological carrier. By doping biodegradable-polymer drug delivery microspheres with natural or synthetic fluorophores, the desired fluorescence can be attained or biological organisms can be simulated without the associated risks and logistical difficulties of live microorganisms.

  13. Cell Based Meniscal Repair Using an Aligned Bioactive Nanofibrous Sheath

    2017-07-01

    to subsequently guide tissue regeneration , for example, by seeded tissue progenitor cells . To achieve this objective, the first step is to develop...AWARD NUMBER: W81XWH-15-1-0104 TITLE: Cell -Based Meniscal Repair Using an Aligned Bioactive Nanofibrous Sheath PRINCIPAL INVESTIGATOR...SUBTITLE 5a. CONTRACT NUMBER Cell -Based Meniscal Repair Using an Aligned Bioactive Nanofibrous Sheath 5b. GRANT NUMBER W81XWH-15-1-0104 5c. PROGRAM

  14. Fluorescent S-layer fusion proteins

    Kainz, B.

    2010-01-01

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

  15. Fluorescence imaging of soybean flavonol isolines

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

    1998-07-01

    region of the spectrum when excited with radiation in the blue region of the spectrum. Thus, green fluorescence emission due to kaempferol glycosides excited by the blue fluorescent compounds with UV excitation (resonance energy excitation) could become a factor in the fluorescence studies of in vivo plants.

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

    Yuan, Shizhuang; Su, Wei; Wang, Enju

    2017-09-01

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

  17. Optimization of fluorescent proteins

    Bindels, D.S.; Goedhart, J.; Hink, M.A.; van Weeren, L.; Joosen, L.; Gadella (jr.), T.W.J.; Engelborghs, Y.; Visser, A.J.W.G.

    2014-01-01

    Nowadays, fluorescent protein (FP) variants have been engineered to fluoresce in all different colors; to display photoswitchable, or photochromic, behavior; or to show yet other beneficial properties that enable or enhance a still growing set of new fluorescence spectroscopy and microcopy

  18. Problems of fluorescent imaging and its solution using nanofluorophores. Part I: Advantages of fluorescent nanoparticles over conventional organic fluorophores

    Zhelev, Z.; Hadjidekov, G.; Zlateva, G.; Spasov, L.; Bakalova, R.

    2011-01-01

    The application of fluorescence in deep-tissue imaging is rapidly expanding in fast several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecules in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With development of novel bright fluorophores based on nano-technologies and fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. This review outlines the current status and future trends of fluorescent nanoparticles - quantum dots (QDs), as a new generation of fluorophores in experimental and pre-clinical fluorescent imaging diagnostic. Part 1 focuses on the advantages of quantum dots over conventional organic fluorophores and defines the major requirements to the 'perfect' fluorophore for fluorescent deep-tissue imaging diagnostic. The analysis is based on the limitations of fluorescent imaging in vivo and overcome by using quantum dots

  19. Nonlinear resonances

    Rajasekar, Shanmuganathan

    2016-01-01

    This introductory text presents the basic aspects and most important features of various types of resonances and anti-resonances in dynamical systems. In particular, for each resonance, it covers the theoretical concepts, illustrates them with case studies, and reviews the available information on mechanisms, characterization, numerical simulations, experimental realizations, possible quantum analogues, applications and significant advances made over the years. Resonances are one of the most fundamental phenomena exhibited by nonlinear systems and refer to specific realizations of maximum response of a system due to the ability of that system to store and transfer energy received from an external forcing source. Resonances are of particular importance in physical, engineering and biological systems - they can prove to be advantageous in many applications, while leading to instability and even disasters in others. The book is self-contained, providing the details of mathematical derivations and techniques invo...

  20. Recent developments in multimodality fluorescence imaging probes

    Jianhong Zhao

    2018-05-01

    Full Text Available Multimodality optical imaging probes have emerged as powerful tools that improve detection sensitivity and accuracy, important in disease diagnosis and treatment. In this review, we focus on recent developments of optical fluorescence imaging (OFI probe integration with other imaging modalities such as X-ray computed tomography (CT, magnetic resonance imaging (MRI, positron emission tomography (PET, single-photon emission computed tomography (SPECT, and photoacoustic imaging (PAI. The imaging technologies are briefly described in order to introduce the strengths and limitations of each techniques and the need for further multimodality optical imaging probe development. The emphasis of this account is placed on how design strategies are currently implemented to afford physicochemically and biologically compatible multimodality optical fluorescence imaging probes. We also present studies that overcame intrinsic disadvantages of each imaging technique by multimodality approach with improved detection sensitivity and accuracy. KEY WORDS: Optical imaging, Fluorescence, Multimodality, Near-infrared fluorescence, Nanoprobe, Computed tomography, Magnetic resonance imaging, Positron emission tomography, Single-photon emission computed tomography, Photoacoustic imaging

  1. Atomic-fluorescence spectrophotometry

    Bakhturova, N.F.; Yudelevich, I.G.

    1975-01-01

    Atomic-fluorescence spectrophotometry, a comparatively new method for the analysis of trace quantities, has developed rapidly in the past ten years. Theoretical and experimental studies by many workers have shown that atomic-fluorescence spectrophotometry (AFS) is capable of achieving a better limit than atomic absorption for a large number of elements. The present review examines briefly the principles of atomic-fluorescence spectrophotometry and the types of fluorescent transition. The excitation sources, flame and nonflame atomizers, used in AFS are described. The limits of detection achieved up to the present, using flame and nonflame methods of atomization are given

  2. Fluorescence of irradiated hydrocarbons

    Gulis, I.G.; Evdokimenko, V.M.; Lapkovskij, M.P.; Petrov, P.T.; Gulis, I.M.; Markevich, S.V.

    1977-01-01

    A visible fluorescence has been found out in γ-irradiated aqueous of carbohydrates. Two bands have been distinguished in fluorescence spectra of the irradiated solution of dextran: a short-wave band lambdasub(max)=140 nm (where lambda is a wave length) at lambdasub(β)=380 nm and a long-wave band with lambdasub(max)=540 nm at lambdasub(β)=430 nm. A similar form of the spectrum has been obtained for irradiated solutions of starch, amylopectin, lowmolecular glucose. It has been concluded that a macromolecule of polysaccharides includes fluorescent centres. A relation between fluorescence and α-oxiketon groups formed under irradiation has been pointed out

  3. Fluorescent Nanoparticle Uptake for Brain Tumor Visualization

    Rachel Tréhin

    2006-04-01

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

  4. Recent Progress on Plasmon-Enhanced Fluorescence

    Dong Jun

    2015-12-01

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

  5. Cell-based therapies for chronic kidney disease

    van Koppen, A.N.

    2013-01-01

    Chronic kidney disease (CKD) may lead to end-stage renal failure, requiring renal replacement strategies. Development of new therapies to reduce progression of CKD is therefore a major global public health target. The aim of this thesis was to investigate whether cell-based therapies have the

  6. Global Distribution of Businesses Marketing Stem Cell-Based Interventions.

    Berger, Israel; Ahmad, Amina; Bansal, Akhil; Kapoor, Tanvir; Sipp, Douglas; Rasko, John E J

    2016-08-04

    A structured search reveals that online marketing of stem-cell-based interventions is skewed toward developed economies including the United States, Ireland, Australia, and Germany. Websites made broad, imprecise therapeutic claims and frequently failed to detail procedures. Widespread marketing poses challenges to regulators, bioethicists, and those seeking realistic hope from therapies. Copyright © 2016 Elsevier Inc. All rights reserved.

  7. Line broadening in multiphoton processes with a resonant intermediate transition

    Wang, C.C.; James, J.V.; Xia, J.

    1983-01-01

    The linewidth of the excitation spectrum for multiphoton ionization is found to be broadened much more severely than the cascade fluorescence originating from the resonant intermediate level. These results are due to the mutual effects of the ionizing and resonating transitions, which are not properly accounted for in perturbative treatments

  8. Resonance Raman study of benzyl radical

    Langkilde, F.W.; Bajdor, K.; Wilbrandt, R.

    1992-01-01

    Time-resolved resonance Raman spectra are obtained of benzyl radicals created by laser flash photolysis of benzylchloride and diphenylacetone in solution. The spectra are obtained in resonance with the intense 2 2A2-1 B-2(2) transition of benzyl. The strong Raman bands are assigned to totally...... symmetric a1 modes. The remaining observed bands are tentatively assigned to fundamental modes of b1, a2, and b2 symmetry, and to overtones and combinations. The resonance Raman spectra are found to be quite different from previous fluorescence spectra of benzyl, and the origins of these differences...

  9. Membranes and Fluorescence microscopy

    Bagatolli, Luis

    2009-01-01

    Fluorescence spectroscopy-based techniques using conventional fluorimeters have been extensively applied since the late 1960s to study different aspects of membrane-related phenomena, i.e., mainly relating to lipid-lipid and lipid-protein (peptide) interactions. Even though fluorescence...

  10. Multimodal fluorescence imaging spectroscopy

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

    2014-01-01

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

  11. The Response of HeLa Cells to Fluorescent NanoDiamond Uptake

    Hemelaar, Simon R; Saspaanithy, Babujhi; L'Hommelet, Severin R M; Perona Martinez, Felipe P; van der Laan, Kiran J; Schirhagl, Romana

    2018-01-01

    Fluorescent nanodiamonds are promising probes for nanoscale magnetic resonance measurements. Their physical properties predict them to have particularly useful applications in intracellular analysis. Before using them in intracellular experiments however, it should be clear whether diamond particles

  12. Multiquark Resonances

    Esposito, A.; Polosa, A.D.

    2016-01-01

    Multiquark resonances are undoubtedly experimentally observed. The number of states and the amount of details on their properties has been growing over the years. It is very recent the discovery of two pentaquarks and the confirmation of four tetraquarks, two of which had not been observed before. We mainly review the theoretical understanding of this sector of particle physics phenomenology and present some considerations attempting a coherent description of the so called X and Z resonances. The prominent problems plaguing theoretical models, like the absence of selection rules limiting the number of states predicted, motivate new directions in model building. Data are reviewed going through all of the observed resonances with particular attention to their common features and the purpose of providing a starting point to further research.

  13. Neuroaesthetic Resonance

    Brooks, Anthony Lewis

    2013-01-01

    Neuroaesthetic Resonance emerged from a mature body of patient- centered gesture-control research investigating non-formal rehabilitation via ICT-enhanced-Art to question ‘Aesthetic Resonance’. Motivating participation, ludic engagement, and augmenting physical motion in non-formal (fun) treatment...... sessions are achieved via adaptive action-analyzed activities. These interactive virtual environments are designed to empower patients’ creative and/or playful expressions via digital feedback stimuli. Unconscious self- pushing of limits result from innate distractive mechanisms offered by the alternative...... the unencumbered motion-to-computer-generated activities - ‘Music Making’, ‘Painting’, ‘Robotic’ and ‘Video Game’ control. A focus of this position paper is to highlight how Aesthetic Resonance, in this context, relates to the growing body of research on Neuroaesthetics to evolve Neuroaesthetic Resonance....

  14. Baryon Resonances

    Oset, E.; Sarkar, S.; Sun Baoxi; Vicente Vacas, M.J.; Ramos, A.; Gonzalez, P.; Vijande, J.; Martinez Torres, A.; Khemchandani, K.

    2010-01-01

    In this talk I show recent results on how many excited baryon resonances appear as systems of one meson and one baryon, or two mesons and one baryon, with the mesons being either pseudoscalar or vectors. Connection with experiment is made including a discussion on old predictions and recent results for the photoproduction of the Λ(1405) resonance, as well as the prediction of one 1/2 + baryon state around 1920 MeV which might have been seen in the γp→K + Λ reaction.

  15. Fluorescent genetic barcoding in mammalian cells for enhanced multiplexing capabilities in flow cytometry.

    Smurthwaite, Cameron A; Hilton, Brett J; O'Hanlon, Ryan; Stolp, Zachary D; Hancock, Bryan M; Abbadessa, Darin; Stotland, Aleksandr; Sklar, Larry A; Wolkowicz, Roland

    2014-01-01

    The discovery of the green fluorescent protein from Aequorea victoria has revolutionized the field of cell and molecular biology. Since its discovery a growing panel of fluorescent proteins, fluorophores and fluorescent-coupled staining methodologies, have expanded the analytical capabilities of flow cytometry. Here, we exploit the power of genetic engineering to barcode individual cells with genes encoding fluorescent proteins. For genetic engineering, we utilize retroviral technology, which allows for the expression of ectopic genetic information in a stable manner in mammalian cells. We have genetically barcoded both adherent and nonadherent cells with different fluorescent proteins. Multiplexing power was increased by combining both the number of distinct fluorescent proteins, and the fluorescence intensity in each channel. Moreover, retroviral expression has proven to be stable for at least a 6-month period, which is critical for applications such as biological screens. We have shown the applicability of fluorescent barcoded multiplexing to cell-based assays that rely themselves on genetic barcoding, or on classical staining protocols. Fluorescent genetic barcoding gives the cell an inherited characteristic that distinguishes it from its counterpart. Once cell lines are developed, no further manipulation or staining is required, decreasing time, nonspecific background associated with staining protocols, and cost. The increasing number of discovered and/or engineered fluorescent proteins with unique absorbance/emission spectra, combined with the growing number of detection devices and lasers, increases multiplexing versatility, making fluorescent genetic barcoding a powerful tool for flow cytometry-based analysis. © 2013 International Society for Advancement of Cytometry.

  16. Ultrafast electrical control of a resonantly driven single photon source

    Cao, Y.; Bennett, A. J.; Ellis, D. J. P.; Shields, A. J.; Farrer, I.; Ritchie, D. A.

    2014-01-01

    We demonstrate generation of a pulsed stream of electrically triggered single photons in resonance fluorescence, by applying high frequency electrical pulses to a single quantum dot in a p-i-n diode under resonant laser excitation. Single photon emission was verified, with the probability of multiple photon emission reduced to 2.8%. We show that despite the presence of charge noise in the emission spectrum of the dot, resonant excitation acts as a “filter” to generate narrow bandwidth photons

  17. 996 RESONANCE November 2013

    IAS Admin

    996. RESONANCE. November 2013. Page 2. 997. RESONANCE. November 2013. Page 3. 998. RESONANCE. November 2013. Page 4. 999. RESONANCE. November 2013. Page 5. 1000. RESONANCE. November 2013. Page 6. 1001. RESONANCE. November 2013. Page 7. 1002. RESONANCE. November 2013 ...

  18. 817 RESONANCE September 2013

    IAS Admin

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  19. 369 RESONANCE April 2016

    IAS Admin

    369. RESONANCE ⎜ April 2016. Page 2. 370. RESONANCE ⎜ April 2016. Page 3. 371. RESONANCE ⎜ April 2016. Page 4. 372. RESONANCE ⎜ April 2016. Page 5. 373. RESONANCE ⎜ April 2016. Page 6. 374. RESONANCE ⎜ April 2016. Page 7. 375. RESONANCE ⎜ April 2016.

  20. Fluorescence spectral studies on interaction of fluorescent probes with Bovine Serum Albumin (BSA)

    Ghosh, Kaushik, E-mail: ghoshfcy@iitr.ac.in; Rathi, Sweety; Arora, Deepshikha

    2016-07-15

    Interaction of 2-(1-(naphthale-1-ylimino)ethyl)phenol (1), 2-methoxy-4-(((4-methoxyphenyl)imino)methyl)phenol (2) and 2-methoxy-4-((naphthalene-1-ylimino)methyl)phenol (3) with Bovine Serum Albumin (BSA) was examined. Fluorescence spectral data were obtained from the probes by varying the concentration of BSA as well as from BSA by varying the concentration of probes. Synchronous fluorescence measurements were performed and binding constants of the probes were calculated. To understand mode of quenching, Stern–Volmer plot, absorption spectral studies and life time measurements were performed. Förster resonance energy transfer (FRET) was also scrutinized. - Highlights: • Schiff bases with pendant phenolato function and interaction with BSA. • Synchronous fluorescence studies and a preferred interaction with tryptophan. • Probable interaction of probes with Trp-213 residue in the hydrophobic cavity. • 1:1 binding stoichiometry of probes and BSA in Benesi–Hildebrand graph.

  1. Synchrobetatron resonances

    Anon.

    1977-01-01

    At the 1975 Particle Accelerator Conference it was reported that a class of resonances were observed in SPEAR II that had not appeared before in SPEAR I. These resonances occur when the betatron oscillation wave numbers ν/sub x/ or ν/sub y/ and the synchrotron wave number ν/sub s/ satisfy the relation (ν/sub x,y/ - mν/sub s/) = 5, with m an integer denoting the m/sup th/ satellite. The main difference between SPEAR II and SPEAR I is the value of ν/sub s/, which in SPEAR II is approximately 0.04, an order of magnitude larger than in SPEAR I. An ad hoc meeting was held at the 1975 Particle Accelerator Conference, where details of the SPEAR II results were presented and various possible mechanisms for producing these resonances were discussed. Later, experiments were performed at SPEAR to identify the mechanism believed to be the most likely explanation. Some of the current experimental knowledge and theoretical views on the source of these resonances are presented

  2. Autostereogram resonators

    Leavey, Sean; Rae, Katherine; Murray, Adam; Courtial, Johannes

    2012-09-01

    Autostereograms, or "Magic Eye" pictures, are repeating patterns designed to give the illusion of depth. Here we discuss optical resonators that create light patterns which, when viewed from a suitable position by a monocular observer, are autostereograms of the three-dimensional shape of one of the mirror surfaces.

  3. Non-blinking quantum dot with a plasmonic nanoshell resonator

    Ji, Botao; Giovanelli, Emerson; Habert, Benjamin; Spinicelli, Piernicola; Nasilowski, Michel; Xu, Xiangzhen; Lequeux, Nicolas; Hugonin, Jean-Paul; Marquier, Francois; Greffet, Jean-Jacques; Dubertret, Benoit

    2015-02-01

    Colloidal semiconductor quantum dots are fluorescent nanocrystals exhibiting exceptional optical properties, but their emission intensity strongly depends on their charging state and local environment. This leads to blinking at the single-particle level or even complete fluorescence quenching, and limits the applications of quantum dots as fluorescent particles. Here, we show that a single quantum dot encapsulated in a silica shell coated with a continuous gold nanoshell provides a system with a stable and Poissonian emission at room temperature that is preserved regardless of drastic changes in the local environment. This novel hybrid quantum dot/silica/gold structure behaves as a plasmonic resonator with a strong Purcell factor, in very good agreement with simulations. The gold nanoshell also acts as a shield that protects the quantum dot fluorescence and enhances its resistance to high-power photoexcitation or high-energy electron beams. This plasmonic fluorescent resonator opens the way to a new family of plasmonic nanoemitters with robust optical properties.

  4. Fluorescence and Spectral Imaging

    Ralph S. DaCosta

    2007-01-01

    Full Text Available Early identification of dysplasia remains a critical goal for diagnostic endoscopy since early discovery directly improves patient survival because it allows endoscopic or surgical intervention with disease localized without lymph node involvement. Clinical studies have successfully used tissue autofluorescence with conventional white light endoscopy and biopsy for detecting adenomatous colonic polyps, differentiating benign hyperplastic from adenomas with acceptable sensitivity and specificity. In Barrett's esophagus, the detection of dysplasia remains problematic because of background inflammation, whereas in the squamous esophagus, autofluorescence imaging appears to be more dependable. Point fluorescence spectroscopy, although playing a crucial role in the pioneering mechanistic development of fluorescence endoscopic imaging, does not seem to have a current function in endoscopy because of its nontargeted sampling and suboptimal sensitivity and specificity. Other point spectroscopic modalities, such as Raman spectroscopy and elastic light scattering, continue to be evaluated in clinical studies, but still suffer the significant disadvantages of being random and nonimaging. A recent addition to the fluorescence endoscopic imaging arsenal is the use of confocal fluorescence endomicroscopy, which provides real-time optical biopsy for the first time. To improve detection of dysplasia in the gastrointestinal tract, a new and exciting development has been the use of exogenous fluorescence contrast probes that specifically target a variety of disease-related cellular biomarkers using conventional fluorescent dyes and novel potent fluorescent nanocrystals (i.e., quantum dots. This is an area of great promise, but still in its infancy, and preclinical studies are currently under way.

  5. Studies of fluorescence and Auger decay following inner-shell photoionization

    Levin, J.C.; Armen, G.B.

    2004-01-01

    Near inner-shell absorption edges, Auger and fluorescence spectra which characterize the first step of a complex cascade process exhibit properties which are well described by radiationless and radiative resonant Raman scattering theory. We present comparisons of our recent data and theory for Auger decay of argon K vacancies, xenon L vacancies, and of fluorescence decay of xenon L vacancies. A theoretical unification of Auger decay and fluorescence decay is presented which clarifies the similarities and differences between the two processes

  6. Fluorescent discharge lamp

    Mukai, E.; Otsuka, H.; Nomi, K.; Honmo, I.

    1982-01-01

    A rapidly illuminating fluorescent lamp 1,200 mm long and 32.5 mm in diameter with an interior conducting strip which is compatible with conventional fixtures and ballasts is described. The fluorescent lamp is composed of a linear glass tube, electrodes sealed at both ends, mercury and raregas sealed in the glass tube, a fluorescent substance clad on the inner walls of the glass tube, and a clad conducting strip extending the entire length of the glass tube in the axial direction on the inner surface of the tube.

  7. Highly thermostable fluorescent proteins

    Bradbury, Andrew M [Santa Fe, NM; Waldo, Geoffrey S [Santa Fe, NM; Kiss, Csaba [Los Alamos, NM

    2011-03-22

    Thermostable fluorescent proteins (TSFPs), methods for generating these and other stability-enhanced proteins, polynucleotides encoding such proteins, and assays and method for using the TSFPs and TSFP-encoding nucleic acid molecules are provided. The TSFPs of the invention show extremely enhanced levels of stability and thermotolerance. In one case, for example, a TSFP of the invention is so stable it can be heated to 99.degree. C. for short periods of time without denaturing, and retains 85% of its fluorescence when heated to 80.degree. C. for several minutes. The invention also provides a method for generating stability-enhanced variants of a protein, including but not limited to fluorescent proteins.

  8. Stem Cell-Based Therapies for Ischemic Stroke

    Lei Hao

    2014-01-01

    Full Text Available In recent years, stem cell-based approaches have attracted more attention from scientists and clinicians due to their possible therapeutical effect on stroke. Animal studies have demonstrated that the beneficial effects of stem cells including embryonic stem cells (ESCs, inducible pluripotent stem cells (iPSCs, neural stem cells (NSCs, and mesenchymal stem cell (MSCs might be due to cell replacement, neuroprotection, endogenous neurogenesis, angiogenesis, and modulation on inflammation and immune response. Although several clinical studies have shown the high efficiency and safety of stem cell in stroke management, mainly MSCs, some issues regarding to cell homing, survival, tracking, safety, and optimal cell transplantation protocol, such as cell dose and time window, should be addressed. Undoubtably, stem cell-based gene therapy represents a novel potential therapeutic strategy for stroke in future.

  9. Global Regulatory Differences for Gene- and Cell-Based Therapies

    Coppens, Delphi G M; De Bruin, Marie L; Leufkens, Hubert G M

    2017-01-01

    Gene- and cell-based therapies (GCTs) offer potential new treatment options for unmet medical needs. However, the use of conventional regulatory requirements for medicinal products to approve GCTs may impede patient access and therapeutic innovation. Furthermore, requirements differ between...... jurisdictions, complicating the global regulatory landscape. We provide a comparative overview of regulatory requirements for GCT approval in five jurisdictions and hypothesize on the consequences of the observed global differences on patient access and therapeutic innovation....

  10. Recent Advances in Stem Cell-Based Therapeutics for Stroke

    Napoli, Eleonora; Borlongan, Cesar V.

    2016-01-01

    Regenerative medicine for central nervous system disorders, including stroke, has challenged the non-regenerative capacity of the brain. Among the many treatment strategies tailored towards repairing the injured brain, stem cell-based therapeutics have been demonstrated as safe and effective in animal models of stroke, and are being tested in limited clinical trials. We address here key lab-to-clinic translational research that relate to efficacy, safety, and mechanism of action underlying st...

  11. Mesenchymal Stem Cell Based Therapy for Prostate Cancer

    2015-11-01

    Montero-Menei, C.; Menei, P. Mesenchymal Stem Cells as Cellular Vehicles for Delivery of Nanoparticles to Brain Tumors. Biomaterials 2010, 31, 8393... Stem Cells : Considerations for Regenerative Medicine Approaches. Tissue Eng. Part B. Rev. 2010, 16, 159–168. 55. Ellem, S. J.; Taylor, R. a.; Furic, L...Award Number: W81XWH-13-1-0304 TITLE: Mesenchymal Stem Cell -Based Therapy for Prostate Cancer PRINCIPAL INVESTIGATOR: John Isaacs CONTRACTING

  12. Cell-based therapeutic strategies for multiple sclerosis

    Scolding, Neil J; Pasquini, Marcelo; Reingold, Stephen C

    2017-01-01

    and none directly promotes repair. Cell-based therapies, including immunoablation followed by autologous haematopoietic stem cell transplantation, mesenchymal and related stem cell transplantation, pharmacologic manipulation of endogenous stem cells to enhance their reparative capabilities......, and transplantation of oligodendrocyte progenitor cells, have generated substantial interest as novel therapeutic strategies for immune modulation, neuroprotection, or repair of the damaged central nervous system in multiple sclerosis. Each approach has potential advantages but also safety concerns and unresolved...

  13. Theory of analytical curves in atomic fluorescence flame spectrometry

    Hooymayers, H.P.

    An explicit expression for the intensity of atomic resonance fluorescence as a function of atomic concentration in a flame is derived under certain idealized conditions. The expression is generally valid for a pure Doppler absorption line profile as well as for a combined Doppler and collisional

  14. Enhancement of cell-based therapeutic angiogenesis using a novel type of injectable scaffolds of hydroxyapatite-polymer nanocomposite microspheres.

    Yohei Mima

    Full Text Available BACKGROUND: Clinical trials demonstrate the effectiveness of cell-based therapeutic angiogenesis in patients with severe ischemic diseases; however, their success remains limited. Maintaining transplanted cells in place are expected to augment the cell-based therapeutic angiogenesis. We have reported that nano-hydroxyapatite (HAp coating on medical devices shows marked cell adhesiveness. Using this nanotechnology, HAp-coated poly(l-lactic acid (PLLA microspheres, named nano-scaffold (NS, were generated as a non-biological, biodegradable and injectable cell scaffold. We investigate the effectiveness of NS on cell-based therapeutic angiogenesis. METHODS AND RESULTS: Bone marrow mononuclear cells (BMNC and NS or control PLLA microspheres (LA were intramuscularly co-implanted into mice ischemic hindlimbs. When BMNC derived from enhanced green fluorescent protein (EGFP-transgenic mice were injected into ischemic muscle, the muscle GFP level in NS+BMNC group was approximate fivefold higher than that in BMNC or LA+BMNC groups seven days after operation. Kaplan-Meier analysis demonstrated that NS+BMNC markedly prevented hindlimb necrosis (P<0.05 vs. BMNC or LA+BMNC. NS+BMNC revealed much higher induction of angiogenesis in ischemic tissues and collateral blood flow confirmed by three-dimensional computed tomography angiography than those of BMNC or LA+BMNC groups. NS-enhanced therapeutic angiogenesis and arteriogenesis showed good correlations with increased intramuscular levels of vascular endothelial growth factor and fibroblast growth factor-2. NS co-implantation also prevented apoptotic cell death of transplanted cells, resulting in prolonged cell retention. CONCLUSION: A novel and feasible injectable cell scaffold potentiates cell-based therapeutic angiogenesis, which could be extremely useful for the treatment of severe ischemic disorders.

  15. Resonance Energy Transfer Molecular Imaging Application in Biomedicine

    NIE Da-hong1,2;TANG Gang-hua1,3

    2016-11-01

    Full Text Available Resonance energy transfer molecular imaging (RETI can markedly improve signal intensity and tissue penetrating capacity of optical imaging, and have huge potential application in the deep-tissue optical imaging in vivo. Resonance energy transfer (RET is an energy transition from the donor to an acceptor that is in close proximity, including non-radiative resonance energy transfer and radiative resonance energy transfer. RETI is an optical imaging technology that is based on RET. RETI mainly contains fluorescence resonance energy transfer imaging (FRETI, bioluminescence resonance energy transfer imaging (BRETI, chemiluminescence resonance energy transfer imaging (CRETI, and radiative resonance energy transfer imaging (RRETI. RETI is the hot field of molecular imaging research and has been widely used in the fields of biology and medicine. This review mainly focuses on RETI principle and application in biomedicine.

  16. Cell-based therapeutic strategies for multiple sclerosis.

    Scolding, Neil J; Pasquini, Marcelo; Reingold, Stephen C; Cohen, Jeffrey A

    2017-11-01

    The availability of multiple disease-modifying medications with regulatory approval to treat multiple sclerosis illustrates the substantial progress made in therapy of the disease. However, all are only partially effective in preventing inflammatory tissue damage in the central nervous system and none directly promotes repair. Cell-based therapies, including immunoablation followed by autologous haematopoietic stem cell transplantation, mesenchymal and related stem cell transplantation, pharmacologic manipulation of endogenous stem cells to enhance their reparative capabilities, and transplantation of oligodendrocyte progenitor cells, have generated substantial interest as novel therapeutic strategies for immune modulation, neuroprotection, or repair of the damaged central nervous system in multiple sclerosis. Each approach has potential advantages but also safety concerns and unresolved questions. Moreover, clinical trials of cell-based therapies present several unique methodological and ethical issues. We summarize here the status of cell-based therapies to treat multiple sclerosis and make consensus recommendations for future research and clinical trials. © The Author (2017). Published by Oxford University Press on behalf of the Guarantors of Brain.

  17. Reviews in fluorescence 2007

    Lakowicz, Joseph R; Geddes, Chris D

    2009-01-01

    This fourth volume in the Springer series summarizes the year's progress in fluorescence, with authoritative analytical reviews specialized enough for professional researchers, yet also appealing to a wider audience of scientists in related fields.

  18. Introduction to fluorescence

    Jameson, David M

    2014-01-01

    "An essential contribution to educating scientists in the principles of fluorescence. It will also be an important addition to the libraries of practitioners applying the principles of molecular fluorescence."-Ken Jacobson, Kenan Distinguished Professor of Cell Biology and Physiology, University of North Carolina at Chapel Hill"An exquisite compendium of fluorescence and its applications in biochemistry enriched by a very exciting historical perspective. This book will become a standard text for graduate students and other scientists."-Drs. Zygmunt (Karol) Gryczynski and Ignacy Gryczynski, University of North Texas Health Science Center"… truly a masterwork, combining clarity, precision, and good humor. The reader, novice or expert, will be pleased with the text and will not stop reading. It is a formidable account of the fluorescence field, which has impacted the life sciences so considerably in the last 60 years."-Jerson L. Silva, M.D., Ph.D., Professor and Director, National Institute of Science and Tech...

  19. Fluorescent magnetic hybrid nanoprobe for multimodal bioimaging

    Koktysh, Dmitry [Department of Chemistry, Vanderbilt University, Station B 351822, Nashville, TN 37235 (United States); Bright, Vanessa; Pham, Wellington, E-mail: dmitry.koktysh@vanderbilt.edu, E-mail: wellington.pham@vanderbilt.edu [Institute of Imaging Science, Vanderbilt University, 1161 21st Avenue South AA, 1105 MCN, Nashville, TN 37232 (United States)

    2011-07-08

    A fluorescent magnetic hybrid imaging nanoprobe (HINP) was fabricated by the conjugation of superparamagnetic Fe{sub 3}O{sub 4} nanoparticles and visible light emitting ({approx}600 nm) fluorescent CdTe/CdS quantum dots (QDs). The assembly strategy used the covalent linking of the oxidized dextran shell of magnetic particles to the glutathione ligands of QDs. The synthesized HINP formed stable water-soluble colloidal dispersions. The structure and properties of the particles were characterized by transmission electron and atomic force microscopy, energy dispersive x-ray analysis and inductively coupled plasma optical emission spectroscopy, dynamic light scattering analysis, optical absorption and photoluminescence spectroscopy, and fluorescent imaging. The luminescence imaging region of the nanoprobe was extended to the near-infrared (NIR) ({approx}800 nm) by conjugation of the superparamagnetic nanoparticles with synthesized CdHgTe/CdS QDs. Cadmium, mercury based QDs in HINP can be easily replaced by novel water-soluble glutathione stabilized AgInS{sub 2}/ZnS QDs to present a new class of cadmium-free multimodal imaging agents. The observed NIR photoluminescence of fluorescent magnetic nanocomposites supports their use for bioimaging. The developed HINP provides dual-imaging channels for simultaneous optical and magnetic resonance imaging.

  20. Fluorescence (Multiwave) Confocal Microscopy.

    Welzel, J; Kästle, Raphaela; Sattler, Elke C

    2016-10-01

    In addition to reflectance confocal microscopy, multiwave confocal microscopes with different laser wavelengths in combination with exogenous fluorophores allow fluorescence mode confocal microscopy in vivo and ex vivo. Fluorescence mode confocal microscopy improves the contrast between the epithelium and the surrounding soft tissue and allows the depiction of certain structures, like epithelial tumors, nerves, and glands. Copyright © 2016 Elsevier Inc. All rights reserved.

  1. Resonating Statements

    Hjelholt, Morten; Jensen, Tina Blegind

    2015-01-01

    IT projects are often complex arrangements of technological components, social actions, and organizational transformation that are difficult to manage in practice. This paper takes an analytical discourse perspective to explore the process of legitimizing IT projects. We introduce the concept...... of resonating statements to highlight how central actors navigate in various discourses over time. Particularly, the statements and actions of an IT project manager are portrayed to show how individuals can legitimize actions by connecting statements to historically produced discourses. The case study...... as part of a feedback loop to re-attach the localized IT project to the broader national discourse. The paper concludes with reflections on how to actively build on resonating statements as a strategic resource for legitimizing IT projects...

  2. Gravitoelectromagnetic resonances

    Tsagas, Christos G.

    2011-01-01

    The interaction between gravitational and electromagnetic radiation has a rather long research history. It is well known, in particular, that gravity-wave distortions can drive propagating electromagnetic signals. Since forced oscillations provide the natural stage for resonances to occur, gravitoelectromagnetic resonances have been investigated as a means of more efficient gravity-wave detection methods. In this report, we consider the coupling between the Weyl and the Maxwell fields on a Minkowski background, which also applies to astrophysical environments where gravity is weak, at the second perturbative level. We use covariant methods that describe gravitational waves via the transverse component of the shear, instead of pure-tensor metric perturbations. The aim is to calculate the properties of the electromagnetic signal, which emerges from the interaction of its linear counterpart with an incoming gravitational wave. Our analysis shows how the wavelength and the amplitude of the gravitationally driven electromagnetic wave vary with the initial conditions. More specifically, for certain initial data, the amplitude of the induced electromagnetic signal is found to diverge. Analogous, diverging, gravitoelectromagnetic resonances were also reported in cosmology. Given that, we extend our Minkowski space study to cosmology and discuss analogies and differences in the physics and in the phenomenology of the Weyl-Maxwell coupling between the aforementioned two physical environments.

  3. Magnetic resonance annual 1986

    Kressel, H.Y.

    1986-01-01

    This book contains papers written on magnetic resonance during 1986. Topics include: musculosketetal magnetic resonance imaging; imaging of the spine; magnetic resonance chemical shift imaging; magnetic resonance imaging in the central nervous system; comparison to computed tomography; high resolution magnetic resonance imaging using surface coils; magnetic resonance imaging of the chest; magnetic resonance imaging of the breast; magnetic resonance imaging of the liver; magnetic resonance spectroscopy of neoplasms; blood flow effects in magnetic resonance imaging; and current and potential applications of clinical sodium magnetic resonance imaging

  4. Fluorescence Image Segmentation by using Digitally Reconstructed Fluorescence Images

    Blumer, Clemens; Vivien, Cyprien; Oertner, Thomas G; Vetter, Thomas

    2011-01-01

    In biological experiments fluorescence imaging is used to image living and stimulated neurons. But the analysis of fluorescence images is a difficult task. It is not possible to conclude the shape of an object from fluorescence images alone. Therefore, it is not feasible to get good manual segmented nor ground truth data from fluorescence images. Supervised learning approaches are not possible without training data. To overcome this issues we propose to synthesize fluorescence images and call...

  5. 1004 RESONANCE November 2013

    IAS Admin

    1004. RESONANCE │ November 2013. Page 2. 1005. RESONANCE │ November 2013. Page 3. 1006. RESONANCE │ November 2013. Page 4. 1007. RESONANCE │ November 2013. Page 5. 1008. RESONANCE │ November 2013. Page 6. 1009. RESONANCE │ November 2013. Page 7. 1010. RESONANCE ...

  6. Even order snake resonances

    Lee, S.Y.

    1993-01-01

    We found that the perturbed spin tune due to the imperfection resonance plays an important role in beam depolarization at snake resonances. We also found that even order snake resonances exist in the overlapping intrinsic and imperfection resonances. Due to the perturbed spin tune shift of imperfection resonances, each snake resonance splits into two

  7. Detecting aromatic compounds on planetary surfaces using ultraviolet time-resolved fluorescence spectroscopy

    Eshelman, E.; Daly, M. G.; Slater, G.; Cloutis, E.

    2018-02-01

    Many aromatic organic molecules exhibit strong and characteristic fluorescence when excited with ultraviolet radiation. As laser excitation in the ultraviolet generates both fluorescence and resonantly enhanced Raman scattering of aromatic vibrational modes, combined Raman and fluorescence instruments have been proposed to search for organic compounds on Mars. In this work the time-resolved fluorescence of a suite of 24 compounds composed of 2-5 ringed alternant, non-alternant, and heterocyclic PAHs was measured. Fluorescence instrumentation with similar specifications to a putative flight instrument was capable of observing the fluorescence decay of these compounds with a sub-ns resolution. Incorporating time-resolved capabilities was also found to increase the ability to discriminate between individual PAHs. Incorporating time-resolved fluorescence capabilities into an ultraviolet gated Raman system intended for a rover or lander can increase the ability to detect and characterize PAHs on planetary surfaces.

  8. Nine New Fluorescent Probes

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

    1989-06-01

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

  9. Sorting live stem cells based on Sox2 mRNA expression.

    Hans M Larsson

    Full Text Available While cell sorting usually relies on cell-surface protein markers, molecular beacons (MBs offer the potential to sort cells based on the presence of any expressed mRNA and in principle could be extremely useful to sort rare cell populations from primary isolates. We show here how stem cells can be purified from mixed cell populations by sorting based on MBs. Specifically, we designed molecular beacons targeting Sox2, a well-known stem cell marker for murine embryonic (mES and neural stem cells (NSC. One of our designed molecular beacons displayed an increase in fluorescence compared to a nonspecific molecular beacon both in vitro and in vivo when tested in mES and NSCs. We sorted Sox2-MB(+SSEA1(+ cells from a mixed population of 4-day retinoic acid-treated mES cells and effectively isolated live undifferentiated stem cells. Additionally, Sox2-MB(+ cells isolated from primary mouse brains were sorted and generated neurospheres with higher efficiency than Sox2-MB(- cells. These results demonstrate the utility of MBs for stem cell sorting in an mRNA-specific manner.

  10. Elastin overexpression by cell-based gene therapy preserves matrix and prevents cardiac dilation

    Li, Shu-Hong; Sun, Zhuo; Guo, Lily; Han, Mihan; Wood, Michael F G; Ghosh, Nirmalya; Alex Vitkin, I; Weisel, Richard D; Li, Ren-Ke

    2012-01-01

    After a myocardial infarction, thinning and expansion of the fibrotic scar contribute to progressive heart failure. The loss of elastin is a major contributor to adverse extracellular matrix remodelling of the infarcted heart, and restoration of the elastic properties of the infarct region can prevent ventricular dysfunction. We implanted cells genetically modified to overexpress elastin to re-establish the elastic properties of the infarcted myocardium and prevent cardiac failure. A full-length human elastin cDNA was cloned, subcloned into an adenoviral vector and then transduced into rat bone marrow stromal cells (BMSCs). In vitro studies showed that BMSCs expressed the elastin protein, which was deposited into the extracellular matrix. Transduced BMSCs were injected into the infarcted myocardium of adult rats. Control groups received either BMSCs transduced with the green fluorescent protein gene or medium alone. Elastin deposition in the infarcted myocardium was associated with preservation of myocardial tissue structural integrity (by birefringence of polarized light; P elastin showed the greatest functional improvement (P elastin in the infarcted heart preserved the elastic structure of the extracellular matrix, which, in turn, preserved diastolic function, prevented ventricular dilation and preserved cardiac function. This cell-based gene therapy provides a new approach to cardiac regeneration. PMID:22435995

  11. Improvement of Bioactive Compound Classification through Integration of Orthogonal Cell-Based Biosensing Methods

    Goran N. Jovanovic

    2007-01-01

    Full Text Available Lack of specificity for different classes of chemical and biological agents, and false positives and negatives, can limit the range of applications for cell-based biosensors. This study suggests that the integration of results from algal cells (Mesotaenium caldariorum and fish chromatophores (Betta splendens improves classification efficiency and detection reliability. Cells were challenged with paraquat, mercuric chloride, sodium arsenite and clonidine. The two detection systems were independently investigated for classification of the toxin set by performing discriminant analysis. The algal system correctly classified 72% of the bioactive compounds, whereas the fish chromatophore system correctly classified 68%. The combined classification efficiency was 95%. The algal sensor readout is based on fluorescence measurements of changes in the energy producing pathways of photosynthetic cells, whereas the response from fish chromatophores was quantified using optical density. Change in optical density reflects interference with the functioning of cellular signal transduction networks. Thus, algal cells and fish chromatophores respond to the challenge agents through sufficiently different mechanisms of action to be considered orthogonal.

  12. A Cell-Based Screen Reveals that the Albendazole Metabolite, Albendazole Sulfone, Targets Wolbachia

    Bray, Walter M.; White, Pamela M.; Ruybal, Jordan; Lokey, R. Scott; Debec, Alain; Sullivan, William

    2012-01-01

    Wolbachia endosymbionts carried by filarial nematodes give rise to the neglected diseases African river blindness and lymphatic filariasis afflicting millions worldwide. Here we identify new Wolbachia-disrupting compounds by conducting high-throughput cell-based chemical screens using a Wolbachia-infected, fluorescently labeled Drosophila cell line. This screen yielded several Wolbachia-disrupting compounds including three that resembled Albendazole, a widely used anthelmintic drug that targets nematode microtubules. Follow-up studies demonstrate that a common Albendazole metabolite, Albendazole sulfone, reduces intracellular Wolbachia titer both in Drosophila melanogaster and Brugia malayi, the nematode responsible for lymphatic filariasis. Significantly, Albendazole sulfone does not disrupt Drosophila microtubule organization, suggesting that this compound reduces titer through direct targeting of Wolbachia. Accordingly, both DNA staining and FtsZ immunofluorescence demonstrates that Albendazole sulfone treatment induces Wolbachia elongation, a phenotype indicative of binary fission defects. This suggests that the efficacy of Albendazole in treating filarial nematode-based diseases is attributable to dual targeting of nematode microtubules and their Wolbachia endosymbionts. PMID:23028321

  13. Genetic barcoding with fluorescent proteins for multiplexed applications.

    Smurthwaite, Cameron A; Williams, Wesley; Fetsko, Alexandra; Abbadessa, Darin; Stolp, Zachary D; Reed, Connor W; Dharmawan, Andre; Wolkowicz, Roland

    2015-04-14

    Fluorescent proteins, fluorescent dyes and fluorophores in general have revolutionized the field of molecular cell biology. In particular, the discovery of fluorescent proteins and their genes have enabled the engineering of protein fusions for localization, the analysis of transcriptional activation and translation of proteins of interest, or the general tracking of individual cells and cell populations. The use of fluorescent protein genes in combination with retroviral technology has further allowed the expression of these proteins in mammalian cells in a stable and reliable manner. Shown here is how one can utilize these genes to give cells within a population of cells their own biosignature. As the biosignature is achieved with retroviral technology, cells are barcoded 'indefinitely'. As such, they can be individually tracked within a mixture of barcoded cells and utilized in more complex biological applications. The tracking of distinct populations in a mixture of cells is ideal for multiplexed applications such as discovery of drugs against a multitude of targets or the activation profile of different promoters. The protocol describes how to elegantly develop and amplify barcoded mammalian cells with distinct genetic fluorescent markers, and how to use several markers at once or one marker at different intensities. Finally, the protocol describes how the cells can be further utilized in combination with cell-based assays to increase the power of analysis through multiplexing.

  14. A Brief Introduction to Single-Molecule Fluorescence Methods.

    van den Wildenberg, Siet M J L; Prevo, Bram; Peterman, Erwin J G

    2018-01-01

    One of the more popular single-molecule approaches in biological science is single-molecule fluorescence microscopy, which will be the subject of the following section of this volume. Fluorescence methods provide the sensitivity required to study biology on the single-molecule level, but they also allow access to useful measurable parameters on time and length scales relevant for the biomolecular world. Before several detailed experimental approaches will be addressed, we will first give a general overview of single-molecule fluorescence microscopy. We start with discussing the phenomenon of fluorescence in general and the history of single-molecule fluorescence microscopy. Next, we will review fluorescent probes in more detail and the equipment required to visualize them on the single-molecule level. We will end with a description of parameters measurable with such approaches, ranging from protein counting and tracking, single-molecule localization super-resolution microscopy, to distance measurements with Förster Resonance Energy Transfer and orientation measurements with fluorescence polarization.

  15. Fluorescence of Alexa fluor dye tracks protein folding.

    Simon Lindhoud

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

  16. Cell-Based Strategies for Meniscus Tissue Engineering

    Niu, Wei; Guo, Weimin; Han, Shufeng; Zhu, Yun; Liu, Shuyun; Guo, Quanyi

    2016-01-01

    Meniscus injuries remain a significant challenge due to the poor healing potential of the inner avascular zone. Following a series of studies and clinical trials, tissue engineering is considered a promising prospect for meniscus repair and regeneration. As one of the key factors in tissue engineering, cells are believed to be highly beneficial in generating bionic meniscus structures to replace injured ones in patients. Therefore, cell-based strategies for meniscus tissue engineering play a fundamental role in meniscal regeneration. According to current studies, the main cell-based strategies for meniscus tissue engineering are single cell type strategies; cell coculture strategies also were applied to meniscus tissue engineering. Likewise, on the one side, the zonal recapitulation strategies based on mimicking meniscal differing cells and internal architectures have received wide attentions. On the other side, cell self-assembling strategies without any scaffolds may be a better way to build a bionic meniscus. In this review, we primarily discuss cell seeds for meniscus tissue engineering and their application strategies. We also discuss recent advances and achievements in meniscus repair experiments that further improve our understanding of meniscus tissue engineering. PMID:27274735

  17. The long-term fate of mesenchymal stem cells labeled with magnetic resonance imaging-visible polymersomes in cerebral ischemia

    Duan X

    2017-09-01

    Full Text Available Xiaohui Duan,1,* Liejing Lu,1,* Yong Wang,2 Fang Zhang,1 Jiaji Mao,1 Minghui Cao,1 Bingling Lin,1 Xiang Zhang,1 Xintao Shuai,2,3 Jun Shen1 1Department of Radiology, Sun Yat-Sen Memorial Hospital, 2PCFM Lab of Ministry of Education, School of Materials Science and Engineering, 3BME Center, Zhongshan School of Medicine, Sun Yat-Sen University, Guangzhou, People’s Republic of China *These authors contributed equally to this work Abstract: Understanding the long-term fate and potential mechanisms of mesenchymal stem cells (MSCs after transplantation is essential for improving functional benefits of stem cell-based stroke treatment. Magnetic resonance imaging (MRI is considered an attractive and clinically translatable tool for longitudinal tracking of stem cells, but certain controversies have arisen in this regard. In this study, we used SPION-loaded cationic polymersomes to label green fluorescent protein (GFP-expressing MSCs to determine whether MRI can accurately reflect survival, long-term fate, and potential mechanisms of MSCs in ischemic stroke therapy. Our results showed that MSCs could improve the functional outcome and reduce the infarct volume of stroke in the brain. In vivo MRI can verify the biodistribution and migration of grafted cells when pre-labeled with SPION-loaded polymersome. The dynamic change of low signal volume on MRI can reflect the tendency of cell survival and apoptosis, but may overestimate long-term survival owing to the presence of iron-laden macrophages around cell graft. Only a small fraction of grafted cells survived up to 8 weeks after transplantation. A minority of these surviving cells were differentiated into astrocytes, but not into neurons. MSCs might exert their therapeutic effect via secreting paracrine factors rather than directing cell replacement through differentiation into neuronal and/or glial phenotypes. Keywords: mesenchymal stem cells, magnetic resonance imaging, superparamagnetic iron oxide

  18. Nanosecond fluorescence spectroscopy

    Leskovar, B.

    1985-03-01

    This article is a summary of a short course lecture given in conjunction with the 1984 Nuclear Science Symposium. Measuring systems for nanosecond fluorescence spectroscopy using single-photon counting techniques are presented. These involve systems based on relaxation-type spark gap light pulser and synchronously pumped mode-locked dye lasers. Furthermore, typical characteristics and optimization of operating conditions of the critical components responsible for the system time resolution are discussed. A short comparison of the most important deconvolution methods for numerical analysis of experimental data is given particularly with respect to the signal-to-noise ratio of the fluorescence signal. 22 refs., 8 figs

  19. Fluorescence uranium determination

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

    1960-01-01

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

  20. Enhancement of single-molecule fluorescence signals by colloidal silver nanoparticles in studies of protein translation.

    Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S; Goldman, Yale E

    2011-01-25

    Metal-enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold, respectively. Fluorescence intensity fluctuations above shot noise, at 0.1-5 Hz, were greater on silver particles. Overall signal-to-noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G.

  1. Enhancement of Single Molecule Fluorescence Signals by Colloidal Silver Nanoparticles in Studies of Protein Translation

    Bharill, Shashank; Chen, Chunlai; Stevens, Benjamin; Kaur, Jaskiran; Smilansky, Zeev; Mandecki, Wlodek; Gryczynski, Ignacy; Gryczynski, Zygmunt; Cooperman, Barry S.; Goldman, Yale E.

    2011-01-01

    Metal enhanced fluorescence (MEF) increased total photon emission of Cy3- and Cy5-labeled ribosomal initiation complexes near 50 nm silver particles 4- and 5.5-fold respectively. Fluorescence intensity fluctuations above shot noise, at 0.1 – 5 Hz, were greater on silver particles. Overall signal to noise ratio was similar or slightly improved near the particles. Proximity to silver particles did not compromise ribosome function, as measured by codon-dependent binding of fluorescent tRNA, dynamics of fluorescence resonance energy transfer between adjacent tRNAs in the ribosome, and tRNA translocation induced by elongation factor G. PMID:21158483

  2. Resonance Fluorescence of Many Interacting Adatoms at a Metal Surface.

    1983-06-01

    we must know the complex function f(d , which can be determined by the Sommerfeld-Hertz vector procedure,2 M 2 1 24 ,+ 2 sp (W p W2 CA) 4 62 {-L...Chem. Phys. 37: 1 (1978). 6. J. H. Eberly, Atomic Relaxation in the Presence of Intense Partially Coherent Radiation Feilds , Phys. Rev. Lett. 37

  3. Potential National Security Applications of Nuclear Resonance Fluorescence Methods

    Warren, Glen A.; Peplowski, Patrick N.; Caggiano, Joseph A.

    2009-01-01

    The objective of this report is to document the initial investigation into the possible research issues related to the development of NRF-based national security applications. The report discusses several potential applications ranging from measuring uranium enrichment in UF6 canisters to characterization of gas samples. While these applications are varied, there are only a few research issues that need to be addressed to understand the limitation of NRF in solving these problems. These research issues range from source and detector development to measuring small samples. The next effort is to determine how best to answer the research issues, followed by a prioritization of those questions to ensure that the most important are addressed. These issues will be addressed through either analytical calculations, computer simulations, analysis of previous data or collection of new measurements. It will also be beneficial to conduct a thorough examination of a couple of the more promising applications in order to develop concrete examples of how NRF may be applied in specific situations. The goals are to develop an understanding of whether the application of NRF is limited by technology or physics in addressing national security applications, to gain a motivation to explore those possible applications, and to develop a research roadmap so that those possibilities may be made reality.

  4. Fluorescence resonance energy transfer (FRET) in chemistry and ...

    Förster distance dependence of the FRET rate. SANGEETA SAINI,1 HARJINDER SINGH2 and BIMAN BAGCHI1,*. 1Solid State and Structural Chemistry Unit, Indian Institute of Science, Bangalore 560 012. 2Permanent address: Department of ...

  5. Distance dependence of fluorescence resonance energy transfer ∑

    Administrator

    eq) is just the electrostatic interaction between the transition ... understanding the conformational dynamics of biological molecules like proteins, RNA, etc. When the donor and ...... where Au is the surface area of the unit cell of the cylindrical ...

  6. Fluorescence imaging as a diagnostic of M-band x-ray drive condition in hohlraum with fluorescent Si targets

    Li, Qi; Hu, Zhimin; Yao, Li; Huang, Chengwu; Yuan, Zheng; Zhao, Yang; Xiong, Gang; Qing, Bo; Lv, Min; Zhu, Tuo; Deng, Bo; Li, Jin; Wei, Minxi; Zhan, Xiayu; Li, Jun; Yang, Yimeng; Su, Chunxiao; Yang, Guohong; Zhang, Jiyan; Li, Sanwei

    2017-01-01

    Fluorescence imaging of surrogate Si-doped CH targets has been used to provide a measurement for drive condition of high-energy x-ray (i.e. M-band x-ray) drive symmetry upon the capsule in hohlraum on Shenguang-II laser facility. A series of experiments dedicated to the study of photo-pumping and fluorescence effect in Si-plasma are presented. To investigate the feasibility of fluorescence imaging in Si-plasma, an silicon plasma in Si-foil target is pre-formed at ground state by the soft x-ray from a half-hohlraum, which is then photo-pumped by the K-shell lines from a spatially distinct laser-produced Si-plasma. The resonant Si photon pump is used to improve the fluorescence signal and cause visible image in the Si-foil. Preliminary fluorescence imaging of Si-ball target is performed in both Si-doped and pure Au hohlraum. The usual capsule at the center of the hohlraum is replaced with a solid Si-doped CH-ball (Si-ball). Since the fluorescence is proportional to the photon pump upon the Si-plasma, high-energy x-ray drive symmetry is equal to the fluorescence distribution of the Si-ball. (paper)

  7. A cell-based fluorescent assay to detect the activity of AB toxins that inhibit protein synthesis

    AB-type protein toxins, produced by numerous bacterial pathogens and some plants, elicit a cytotoxic effect involving the inhibition of protein synthesis. To develop an improved method to detect the inhibition of protein synthesis by AB-type toxins, the present study characterized a Vero cell line t...

  8. An Experimental Study of the Fluorescence Spectrum of Cesium Atoms in the Presence of a Buffer Gas

    Davydov, V. G.; Kulyasov, V. N.

    2018-01-01

    A direct experiment is performed to determine the quantum efficiency of a cesium fluorescence filter. The fluorescence spectra of cesium atoms are recorded under excitation of the upper states of the second resonance doublet with a Bell-Bloom cesium lamp. Introduction of different noble gases into the cell with cesium leads to the appearance of additional fluorescence photons. It is found that a fluorescence filter based on atomic cesium vapor with addition of helium in the working cell has the highest efficiency and response rate of all known fluorescence filters based on alkali-metal atomic vapors.

  9. Monitoring by fluorescence measurements

    Malcolme-Lawes, D.J.; Gifford, L.A.

    1981-01-01

    A fluorimetric detector is described in which the fluorescence excitation source may be 3 H, 14 C, 35 S, 147 Pm or 63 Ni. Such a detector can be adapted for use with flowing liquid systems especially liquid chromatography systems. (U.K.)

  10. Fluorescence lifetime based bioassays

    Meyer-Almes, Franz-Josef

    2017-12-01

    Fluorescence lifetime (FLT) is a robust intrinsic property and material constant of fluorescent matter. Measuring this important physical indicator has evolved from a laboratory curiosity to a powerful and established technique for a variety of applications in drug discovery, medical diagnostics and basic biological research. This distinct trend was mainly driven by improved and meanwhile affordable laser and detection instrumentation on the one hand, and the development of suitable FLT probes and biological assays on the other. In this process two essential working approaches emerged. The first one is primarily focused on high throughput applications employing biochemical in vitro assays with no requirement for high spatial resolution. The second even more dynamic trend is the significant expansion of assay methods combining highly time and spatially resolved fluorescence data by fluorescence lifetime imaging. The latter approach is currently pursued to enable not only the investigation of immortal tumor cell lines, but also specific tissues or even organs in living animals. This review tries to give an actual overview about the current status of FLT based bioassays and the wide range of application opportunities in biomedical and life science areas. In addition, future trends of FLT technologies will be discussed.

  11. Fluorescent Lamp Replacement Study

    2017-07-01

    not be cited for purposes of advertisement. DISPOSITION INSTRUCTIONS: Destroy this document when no longer needed. Do not return to the... recycling , and can be disposed safely in a landfill. (2) LEDs offer reduced maintenance costs and fewer bulb replacements, significantly reducing... recycling . Several fixtures, ballasts and energy efficient fluorescent bulbs that were determined to be in pristine condition were returned to ATC

  12. Applied neutron resonance theory

    Froehner, F.H.

    1980-01-01

    Utilisation of resonance theory in basic and applications-oriented neutron cross section work is reviewed. The technically important resonance formalisms, principal concepts and methods as well as representative computer programs for resonance parameter extraction from measured data, evaluation of resonance data, calculation of Doppler-broadened cross sections and estimation of level-statistical quantities from resonance parameters are described. (author)

  13. Applied neutron resonance theory

    Froehner, F.H.

    1978-07-01

    Utilisation of resonance theory in basic and applications-oriented neutron cross section work is reviewed. The technically important resonance formalisms, principal concepts and methods as well as representative computer programs for resonance parameter extraction from measured data, evaluation of resonance data, calculation of Doppler-broadened cross sections and estimation of level-statistical quantities from resonance parameters are described. (orig.) [de

  14. Statistical filtering in fluorescence microscopy and fluorescence correlation spectroscopy

    Macháň, Radek; Kapusta, Peter; Hof, Martin

    Roč. 406 , č. 20 (2014), s. 4797-4813 ISSN 1618-2642 R&D Projects: GA ČR GBP208/12/G016 Institutional support: RVO:61388955 Keywords : Filtered fluorescence correlation spectroscopy * Fluorescence lifetime correlation spectroscopy * Fluorescence spectral correlation spectroscopy Subject RIV: CF - Physical ; Theoretical Chemistry Impact factor: 3.436, year: 2014

  15. Cooperative fluorescence from a strongly driven dilute cloud of atoms

    Ott, Johan Raunkjær; Wubs, Martijn; Lodahl, Peter

    2013-01-01

    We investigate cooperative fluorescence in a dilute cloud of strongly driven two-level emitters. Starting from the Heisenberg equations of motion, we compute the first-order scattering corrections to the saturation of the excited-state population and to the resonance-fluorescence spectrum, which...... both require going beyond the state-of-the-art linear-optics approach to describe collective phenomena. A dipole blockade is observed due to long-range dipole-dipole coupling that vanishes at stronger driving fields. Furthermore, we compute the inelastic component of the light scattered by a cloud...

  16. New fluorescent probes for detection and characterization of amyloid fibrils

    Gorbenko, Galyna; Trusova, Valeriya; Kirilova, Elena; Kirilov, Georgiy; Kalnina, Inta; Vasilev, Aleksey; Kaloyanova, Stefka; Deligeorgiev, Todor

    2010-08-01

    The applicability of the novel fluorescent probes, aminoderivative of benzanthrone ABM, squaraine dye SQ-1 and polymethine dye V2 to identification and structural analysis of amyloid fibrils has been evaluated using the lysozyme model system in which fibrillar aggregates have been formed in concentrated ethanol solution. The association constant, binding stoichiometry and molar fluorescence of the bound dye have been determined. ABM was found to surpass classical amyloid marker ThT in the sensitivity to the presence of fibrillar aggregates. Resonance energy transfer measurements involving ABM-SQ-1 and SQ-1-V2 donor-acceptor pairs yielded the limits for fractal-like dimension of lysozyme fibrils.

  17. Development of a novel cell-based assay system EPISSAY for screening epigenetic drugs and liposome formulated decitabine

    Lim, Sue Ping; Callen, David F; Kumar, Raman; Akkamsetty, Yamini; Wang, Wen; Ho, Kristen; Neilsen, Paul M; Walther, Diego J; Suetani, Rachel J; Prestidge, Clive

    2013-01-01

    Despite the potential of improving the delivery of epigenetic drugs, the subsequent assessment of changes in their epigenetic activity is largely dependent on the availability of a suitable and rapid screening bioassay. Here, we describe a cell-based assay system for screening gene reactivation. A cell-based assay system (EPISSAY) was designed based on a silenced triple-mutated bacterial nitroreductase TMnfsB fused with Red-Fluorescent Protein (RFP) expressed in the non-malignant human breast cell line MCF10A. EPISSAY was validated using the target gene TXNIP, which has previously been shown to respond to epigenetic drugs. The potency of a epigenetic drug model, decitabine, formulated with PEGylated liposomes was also validated using this assay system. Following treatment with DNA methyltransferase (DNMT) and histone deacetylase (HDAC) inhibitors such as decitabine and vorinostat, increases in RFP expression were observed, indicating expression of RFP-TMnfsB. The EPISSAY system was then used to test the potency of decitabine, before and after PEGylated liposomal encapsulation. We observed a 50% higher potency of decitabine when encapsulated in PEGylated liposomes, which is likely to be due to its protection from rapid degradation. The EPISSAY bioassay system provides a novel and rapid system to compare the efficiencies of existing and newly formulated drugs that reactivate gene expression

  18. Laser induced fluorescence spectroscopy for FTU

    Hughes, T.P.

    1995-07-01

    Laser induced fluorescence spectroscopy (LIFS) is based on the absorption of a short pulse of tuned laser light by a group of atoms and the observation of the resulting fluorescence radiation from the excited state. Because the excitation is resonant it is very efficient, and the fluorescence can be many times brighter than the normal spontaneous emission, so low number densities of the selected atoms can be detected and measured. Good spatial resolution can be achieved by using a narrow laser beam. If the laser is sufficiently monochromatic, and it can be tuned over the absorption line profile of the selected atoms, information can also be obtained about the velocities of the atoms from the Doppler effect which can broaden and shift the line. In this report two topics are examined in detail. The first is the effect of high laser irradiance, which can cause 'power broadening' of the apparent absorption line profile. The second is the effect of the high magnetic field in FTU. Detailed calculations are given for LIFS of neutral iron and molybdenum atoms, including the Zeeman effect, and the implementation of LIFS for these atoms on FTU is discussed

  19. High frequency electromechanical memory cells based on telescoping carbon nanotubes.

    Popov, A M; Lozovik, Y E; Kulish, A S; Bichoutskaia, E

    2010-07-01

    A new method to increase the operational frequency of electromechanical memory cells based on the telescoping motion of multi-walled carbon nanotubes through the selection of the form of the switching voltage pulse is proposed. The relative motion of the walls of carbon nanotubes can be controlled through the shape of the interwall interaction energy surface. This allows the use of the memory cells in nonvolatile or volatile regime, depending on the structure of carbon nanotube. Simulations based on ab initio and semi-empirical calculations of the interwall interaction energies are used to estimate the switching voltage and the operational frequency of volatile cells with the electrodes made of carbon nanotubes. The lifetime of nonvolatile memory cells is also predicted.

  20. Methods and practices to diversify cell-based products.

    Vertès, Alain A

    2017-12-15

    Medicinal signaling cell (MSC)-based products represent emerging treatments in various therapeutic areas including cardiometabolic, inflammation, autoimmunity, orthopedics, wound healing and oncology. Exploring innovation beyond minimally manipulated plastic-adherent ex vivo expanded allogeneic MSCs enables product delineation. Product delineation is on the critical path to maximize clinical benefits and market access. An innovation framework is presented here along various innovation dimensions comprising composition-of-matter by means of positive cell surface markers, formulation varying for example the cell dose or the preservation mode and medium, manufacturing to adapt the secretome of MSCs to the condition of interest, the mode of delivery and corresponding delivery devices, as well as molecular engineering and biomarkers. The rationale of the innovation space thus described applies generally to all cell-based therapies.

  1. Glial progenitor cell-based treatment of the childhood leukodystrophies

    Osório, M. Joana; Goldman, Steven A.

    2016-01-01

    stem cell-derived human neural or glial progenitor cells may comprise a promising strategy for both structural remyelination and metabolic rescue. A broad variety of pediatric white matter disorders, including the primary hypomyelinating disorders, the lysosomal storage disorders, and the broader group...... genetic editing of pluripotent stem cells. Yet these challenges notwithstanding, the promise of glial progenitor cell-based treatment of the childhood myelin disorders offers hope to the many victims of this otherwise largely untreatable class of disease....... and astrocytes are the major affected cell populations, and are either structurally impaired or metabolically compromised through cell-intrinsic pathology, or are the victims of mis-accumulated toxic byproducts of metabolic derangement. In either case, glial cell replacement using implanted tissue or pluripotent...

  2. Cell-Based Microarrays for In Vitro Toxicology

    Wegener, Joachim

    2015-07-01

    DNA/RNA and protein microarrays have proven their outstanding bioanalytical performance throughout the past decades, given the unprecedented level of parallelization by which molecular recognition assays can be performed and analyzed. Cell microarrays (CMAs) make use of similar construction principles. They are applied to profile a given cell population with respect to the expression of specific molecular markers and also to measure functional cell responses to drugs and chemicals. This review focuses on the use of cell-based microarrays for assessing the cytotoxicity of drugs, toxins, or chemicals in general. It also summarizes CMA construction principles with respect to the cell types that are used for such microarrays, the readout parameters to assess toxicity, and the various formats that have been established and applied. The review ends with a critical comparison of CMAs and well-established microtiter plate (MTP) approaches.

  3. Characterisation of a fuel cell based uninteruptable power supply

    Aklil, D.; Gazey, R.; McGrath, D.

    2004-07-01

    This report presents the findings of tests carried out to determine if a fuel cell (FC) could be used instead of external batteries in UPS systems. Details are given of the configuration of the 1kW fuel cell based test UPS system (FC-UPS), fuel cell suitability for UPS, the start-up conditions, the on-load dynamic response, comparative weight/space savings of FC-UPS, lifetime costs compared to battery installations, and market readiness of FC systems for UPS deployment. The importance of the collaboration between the FC manufacturers and system integrator for the implementation of the project and of the testing and characterisation of FC products is stressed.

  4. [Proangiogenic cell-based therapy for treatment of ischemic diseases].

    Silvestre, Jean-Sébastien

    2009-11-01

    The application of endothelial progenitor cells (EPC) cell-based therapy for regenerative medicine constitutes a promising therapeutic avenue for the treatment of cardiovascular diseases. Based on experimental studies demonstrating that bone marrow-, blood- or tissue-derived stem/progenitor cells improve the functional recovery after ischemia, clinical trials were initiated to address this new therapeutic concept. Although autolougous cell therapy was shown to improve perfusion and function of ischemic tissues, a number of issues remain to be adressed. The nature of the mobilizing, migratory and homing signals, and the mechanisms of action need to be identified and further defined. In addition, strategies to enhance homing, survival and therapeutic potential of EPC need to be developped to improve therapeutic effect and counteract EPC dysfunction in aged patients with cardiovascular risk factors. The present review article will discuss the mechanisms of action of different types of adult stem cells and several approaches to improve their therapeutic efficiency.

  5. Printing technologies for biomolecule and cell-based applications.

    Ihalainen, Petri; Määttänen, Anni; Sandler, Niklas

    2015-10-30

    Biomolecules, such as enzymes, proteins and other biomacromolecules (polynucleotides, polypeptides, polysaccharides and DNA) that are immobilized on solid surfaces are relevant to many areas of science and technology. These functionalized surfaces have applications in biosensors, chromatography, diagnostic immunoassays, cell culturing, DNA microarrays and other analytical techniques. Printing technologies offer opportunities in this context. The main interests in printing biomolecules are in immobilizing them on surfaces for sensors and catalysts or for controlled delivery of protein-based drugs. Recently, there have been significant developments in the use of inkjet printing for dispensing of proteins, biomacromolecules and cells. This review discusses the use of roll-to-roll and inkjet printing technologies in manufacturing of biomolecule and cell-based applications. Copyright © 2015. Published by Elsevier B.V.

  6. Superthin Solar Cells Based on AIIIBV/Ge Heterostructures

    Pakhanov, N. A.; Pchelyakov, O. P.; Vladimirov, V. M.

    2017-11-01

    A comparative analysis of the prospects of creating superthin, light-weight, and highly efficient solar cells based on AIIIBV/InGaAs and AIIIBV/Ge heterostructures is performed. Technological problems and prospects of each variant are discussed. A method of thinning of AIIIBV/Ge heterostructures with the use of an effective temporary carrier is proposed. The method allows the process to be performed almost with no risk of heterostructure fracture, thinning of the Ge junction down to several tens of micrometers (or even several micrometers), significant enhancement of the yield of good structures, and also convenient and reliable transfer of thinned solar cells to an arbitrary light and flexible substrate. Such a technology offers a possibility of creating high-efficiency thin and light solar cells for space vehicles on the basis of mass-produced AIIIBV/Ge heterostructures.

  7. Microencapsulating and Banking Living Cells for Cell-Based Medicine

    Wujie Zhang

    2011-01-01

    Full Text Available A major challenge to the eventual success of the emerging cell-based medicine such as tissue engineering, regenerative medicine, and cell transplantation is the limited availability of the desired cell sources. This challenge can be addressed by cell microencapsulation to overcome the undesired immune response (i.e., to achieve immunoisolation so that non-autologous cells can be used to treat human diseases, and by cell/tissue preservation to bank living cells for wide distribution to end users so that they are readily available when needed in the future. This review summarizes the status quo of research in both cell microencapsulation and banking the microencapsulated cells. It is concluded with a brief outlook of future research directions in this important field.

  8. Synthetic biology in cell-based cancer immunotherapy.

    Chakravarti, Deboki; Wong, Wilson W

    2015-08-01

    The adoptive transfer of genetically engineered T cells with cancer-targeting receptors has shown tremendous promise for eradicating tumors in clinical trials. This form of cellular immunotherapy presents a unique opportunity to incorporate advanced systems and synthetic biology approaches to create cancer therapeutics with novel functions. We first review the development of synthetic receptors, switches, and circuits to control the location, duration, and strength of T cell activity against tumors. In addition, we discuss the cellular engineering and genome editing of host cells (or the chassis) to improve the efficacy of cell-based cancer therapeutics, and to reduce the time and cost of manufacturing. Copyright © 2015 Elsevier Ltd. All rights reserved.

  9. Cell based assays for anti-Plasmodium activity evaluation.

    Mokgethi-Morule, Thabang; N'Da, David D

    2016-03-10

    Malaria remains one of the most common and deadly infectious diseases worldwide. The severity of this global public health challenge is reflected by the approximately 198 million people, who were reportedly infected in 2013 and by the more than 584,000 related deaths in that same year. The rising emergence of drug resistance towards the once effective artemisinin combination therapies (ACTs) has become a serious concern and warrants more robust drug development strategies, with the objective of eradicating malaria infections. The intricate biology and life cycle of Plasmodium parasites complicate the understanding of the disease in such a way that would enhance the development of more effective chemotherapies that would achieve radical clinical cure and that would prevent disease relapse. Phenotypic cell based assays have for long been a valuable approach and involve the screening and analysis of diverse compounds with regards to their activities towards whole Plasmodium parasites in vitro. To achieve the Millennium Development Goal (MDG) of malaria eradication by 2020, new generation drugs that are active against all parasite stages (erythrocytic (blood), exo-erythrocytic (liver stages and gametocytes)) are needed. Significant advances are being made in assay development to overcome some of the practical challenges of assessing drug efficacy, particularly in the liver and transmission stage Plasmodium models. This review discusses primary screening models and the fundamental progress being made in whole cell based efficacy screens of anti-malarial activity. Ongoing challenges and some opportunities for improvements in assay development that would assist in the discovery of effective, safe and affordable drugs for malaria treatments are also discussed. Copyright © 2016 Elsevier B.V. All rights reserved.

  10. Who's who in fluorescence 2008

    Geddes, Chris D

    2008-01-01

    The Journal of Fluorescence's sixth Who's Who directory publishes the names, contact details, specialty keywords, and a brief description of scientists employing fluorescence methodology and instrumentation in their working lives. This is a unique reference.

  11. Quantum dots versus organic fluorophores in fluorescent deep-tissue imaging--merits and demerits.

    Bakalova, Rumiana; Zhelev, Zhivko; Gadjeva, Veselina

    2008-12-01

    The use of fluorescence in deep-tissue imaging is rapidly expanding in last several years. The progress in fluorescent molecular probes and fluorescent imaging techniques gives an opportunity to detect single cells and even molecular targets in live organisms. The highly sensitive and high-speed fluorescent molecular sensors and detection devices allow the application of fluorescence in functional imaging. With the development of novel bright fluorophores based on nanotechnologies and 3D fluorescence scanners with high spatial and temporal resolution, the fluorescent imaging has a potential to become an alternative of the other non-invasive imaging techniques as magnetic resonance imaging, positron-emission tomography, X-ray, computing tomography. The fluorescent imaging has also a potential to give a real map of human anatomy and physiology. The current review outlines the advantages of fluorescent nanoparticles over conventional organic dyes in deep-tissue imaging in vivo and defines the major requirements to the "perfect fluorophore". The analysis proceeds from the basic principles of fluorescence and major characteristics of fluorophores, light-tissue interactions, and major limitations of fluorescent deep-tissue imaging. The article is addressed to a broad readership - from specialists in this field to university students.

  12. Narrow dibaryon resonances

    Kajdalov, A.B.

    1986-01-01

    Experimental data on np interactions indicating to existence of narrow resonances in pp-system are discussed. Possible theoretical interpretations of these resonances are given. Experimental characteristics of the dibaryon resonances with isospin I=2 are considered

  13. MRI (Magnetic Resonance Imaging)

    ... Procedures Medical Imaging MRI (Magnetic Resonance Imaging) MRI (Magnetic Resonance Imaging) Share Tweet Linkedin Pin it More sharing options Linkedin Pin it Email Print Magnetic Resonance Imaging (MRI) is a medical imaging procedure for ...

  14. Synthesis and Fluorescence Spectra of Triazolylcoumarin Fluorescent Dyes

    PENG Xian-fu; LI Hong-qi

    2009-01-01

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

  15. Fluorescence spectroscopy of dental calculus

    Bakhmutov, D; Gonchukov, S; Sukhinina, A

    2010-01-01

    The aim of the present study was to investigate the fluorescence properties of dental calculus in comparison with the properties of adjacent unaffected tooth structure using both lasers and LEDs in the UV-visible range for fluorescence excitation. The influence of calculus color on the informative signal is demonstrated. The optimal spectral bands of excitation and registration of the fluorescence are determined

  16. Fluorescence spectroscopy of dental calculus

    Bakhmutov, D.; Gonchukov, S.; Sukhinina, A.

    2010-05-01

    The aim of the present study was to investigate the fluorescence properties of dental calculus in comparison with the properties of adjacent unaffected tooth structure using both lasers and LEDs in the UV-visible range for fluorescence excitation. The influence of calculus color on the informative signal is demonstrated. The optimal spectral bands of excitation and registration of the fluorescence are determined.

  17. Fluorescence Imaging Reveals Surface Contamination

    Schirato, Richard; Polichar, Raulf

    1992-01-01

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

  18. Who's who in fluorescence 2005

    Geddes, Chris D

    2006-01-01

    The Journal of Fluorescence's third Who's Who directory publishes the names, contact details, specialty keywords, photographs, and a brief description of scientists employing fluorescence methodology and instrumentation in their working livesThe directory provides company contact details with a brief list of fluorescence-related products.

  19. Regenerative feedback resonant circuit

    Jones, A. Mark; Kelly, James F.; McCloy, John S.; McMakin, Douglas L.

    2014-09-02

    A regenerative feedback resonant circuit for measuring a transient response in a loop is disclosed. The circuit includes an amplifier for generating a signal in the loop. The circuit further includes a resonator having a resonant cavity and a material located within the cavity. The signal sent into the resonator produces a resonant frequency. A variation of the resonant frequency due to perturbations in electromagnetic properties of the material is measured.

  20. Characterization of a spectrally diverse set of fluorescent proteins as FRET acceptors for mTurquoise2

    Mastop, M.; Bindels, D.S.; Shaner, N.C.; Postma, M.; Gadella, T.W.J.; Goedhart, J.

    2017-01-01

    The performance of Förster Resonance Energy Transfer (FRET) biosensors depends on brightness and photostability, which are dependent on the characteristics of the fluorescent proteins that are employed. Yellow fluorescent protein (YFP) is often used as an acceptor but YFP is prone to photobleaching

  1. Imaging molecular interactions in cells by dynamic and static fluorescence anisotropy (rFLIM and emFRET)

    Lidke, D.S.; Nagy, P.; Barisas, B.G.; Heintzmann, R.; Post, Janine Nicole; Lidke, K.A.; Clayton, A.H.A.; Arndt-jovin, D.J.; Jovin, T.M.

    2003-01-01

    We report the implementation and exploitation of fluorescence polarization measurements, in the form of anisotropy fluorescence lifetime imaging microscopy (rFLIM) and energy migration Förster resonance energy transfer (emFRET) modalities, for wide-field, confocal laser-scanning microscopy and flow

  2. Synthesis and bio-applications of targeted magnetic-fluorescent composite nanoparticles

    Xia, Hui; Tong, Ruijie; Song, Yanling; Xiong, Fang; Li, Jiman; Wang, Shichao; Fu, Huihui; Wen, Jirui; Li, Dongze; Zeng, Ye; Zhao, Zhiwei; Wu, Jiang

    2017-01-01

    Magnetic-fluorescent nanoparticles have a tremendous potential in biology. As the benefits of these materials gained recognition, increasing attention has been given to the conjugation of magnetic-fluorescent nanoparticles with targeting ligands. The magnetic and fluorescent properties of nanoparticles offer several functionalities, including imaging, separation, and visualization, while the presence of a targeting ligand allows for selective cell and tissue targeting. In this review, methods for the synthesis of targeted magnetic-fluorescent nanoparticles are explored, and recent applications of these nanocomposites to the detection and separation of biomolecules, fluorescent and magnetic resonance imaging, and cancer diagnosis and treatment will be summarized. As these materials are further optimized, targeted magnetic-fluorescent nanoparticles hold great promise for the diagnosis and treatment of some diseases.

  3. Synthesis and bio-applications of targeted magnetic-fluorescent composite nanoparticles

    Xia, Hui; Tong, Ruijie; Song, Yanling; Xiong, Fang; Li, Jiman; Wang, Shichao; Fu, Huihui; Wen, Jirui; Li, Dongze; Zeng, Ye; Zhao, Zhiwei; Wu, Jiang

    2017-04-01

    Magnetic-fluorescent nanoparticles have a tremendous potential in biology. As the benefits of these materials gained recognition, increasing attention has been given to the conjugation of magnetic-fluorescent nanoparticles with targeting ligands. The magnetic and fluorescent properties of nanoparticles offer several functionalities, including imaging, separation, and visualization, while the presence of a targeting ligand allows for selective cell and tissue targeting. In this review, methods for the synthesis of targeted magnetic-fluorescent nanoparticles are explored, and recent applications of these nanocomposites to the detection and separation of biomolecules, fluorescent and magnetic resonance imaging, and cancer diagnosis and treatment will be summarized. As these materials are further optimized, targeted magnetic-fluorescent nanoparticles hold great promise for the diagnosis and treatment of some diseases.

  4. Synthesis and bio-applications of targeted magnetic-fluorescent composite nanoparticles

    Xia, Hui; Tong, Ruijie [Sichuan University, West China Medical Center (China); Song, Yanling [Shenyang University of Chemical Technology, College of Pharmaceutical and Biological Engineering (China); Xiong, Fang [Sichuan University, West China College of Stomatology (China); Li, Jiman [Sichuan Cancer Hospital, Pathology Department (China); Wang, Shichao; Fu, Huihui; Wen, Jirui; Li, Dongze; Zeng, Ye; Zhao, Zhiwei, E-mail: zzw2002400@126.com; Wu, Jiang, E-mail: jw@scu.edu.cn [Sichuan University, West China Medical Center (China)

    2017-04-15

    Magnetic-fluorescent nanoparticles have a tremendous potential in biology. As the benefits of these materials gained recognition, increasing attention has been given to the conjugation of magnetic-fluorescent nanoparticles with targeting ligands. The magnetic and fluorescent properties of nanoparticles offer several functionalities, including imaging, separation, and visualization, while the presence of a targeting ligand allows for selective cell and tissue targeting. In this review, methods for the synthesis of targeted magnetic-fluorescent nanoparticles are explored, and recent applications of these nanocomposites to the detection and separation of biomolecules, fluorescent and magnetic resonance imaging, and cancer diagnosis and treatment will be summarized. As these materials are further optimized, targeted magnetic-fluorescent nanoparticles hold great promise for the diagnosis and treatment of some diseases.

  5. Bright and photostable nitrogen-vacancy fluorescence from unprocessed detonation nanodiamond.

    Reineck, P; Capelli, M; Lau, D W M; Jeske, J; Field, M R; Ohshima, T; Greentree, A D; Gibson, B C

    2017-01-05

    Bright and photostable fluorescence from nitrogen-vacancy (NV) centers is demonstrated in unprocessed detonation nanodiamond particle aggregates. The optical properties of these particles is analyzed using confocal fluorescence microscopy and spectroscopy, time resolved fluorescence decay measurements, and optically detected magnetic resonance experiments. Two particle populations with distinct optical properties are identified and compared to high-pressure high-temperature (HPHT) fluorescent nanodiamonds. We find that the brightness of one detonation nanodiamond particle population is on the same order as that of highly processed fluorescent 100 nm HPHT nanodiamonds. Our results may open the path to a simple and up-scalable route for the production of fluorescent NV nanodiamonds for use in bioimaging applications.

  6. Resonances, resonance functions and spectral deformations

    Balslev, E.

    1984-01-01

    The present paper is aimed at an analysis of resonances and resonance states from a mathematical point of view. Resonances are characterized as singular points of the analytically continued Lippman-Schwinger equation, as complex eigenvalues of the Hamiltonian with a purely outgoing, exponentially growing eigenfunction, and as poles of the S-matrix. (orig./HSI)

  7. The controversial origin of pericytes during angiogenesis - Implications for cell-based therapeutic angiogenesis and cell-based therapies.

    Blocki, Anna; Beyer, Sebastian; Jung, Friedrich; Raghunath, Michael

    2018-01-01

    Pericytes reside within the basement membrane of small vessels and are often in direct cellular contact with endothelial cells, fulfilling important functions during blood vessel formation and homeostasis. Recently, these pericytes have been also identified as mesenchymal stem cells. Mesenchymal stem cells, and especially their specialized subpopulation of pericytes, represent promising candidates for therapeutic angiogenesis applications, and have already been widely applied in pre-clinical and clinical trials. However, cell-based therapies of ischemic diseases (especially of myocardial infarction) have not resulted in significant long-term improvement. Interestingly, pericytes from a hematopoietic origin were observed in embryonic skin and a pericyte sub-population expressing leukocyte and monocyte markers was described during adult angiogenesis in vivo. Since mesenchymal stem cells do not express hematopoietic markers, the latter cell type might represent an alternative pericyte population relevant to angiogenesis. Therefore, we sourced blood-derived angiogenic cells (BDACs) from monocytes that closely resembled hematopoietic pericytes, which had only been observed in vivo thus far. BDACs displayed many pericytic features and exhibited enhanced revascularization and functional tissue regeneration in a pre-clinical model of critical limb ischemia. Comparison between BDACs and mesenchymal pericytes indicated that BDACs (while resembling hematopoietic pericytes) enhanced early stages of angiogenesis, such as endothelial cell sprouting. In contrast, mesenchymal pericytes were responsible for blood vessel maturation and homeostasis, while reducing endothelial sprouting.Since the formation of new blood vessels is crucial during therapeutic angiogenesis or during integration of implants into the host tissue, hematopoietic pericytes (and therefore BDACs) might offer an advantageous addition or even an alternative for cell-based therapies.

  8. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    Gartia, Manas Ranjan; Hsiao, Austin; Logan Liu, G; Sivaguru, Mayandi; Chen Yi

    2011-01-01

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  9. Enhanced 3D fluorescence live cell imaging on nanoplasmonic substrate

    Gartia, Manas Ranjan [Department of Nuclear, Plasma and Radiological Engineering, University of Illinois, Urbana, IL 61801 (United States); Hsiao, Austin; Logan Liu, G [Department of Bioengineering, University of Illinois, Urbana, IL 61801 (United States); Sivaguru, Mayandi [Institute for Genomic Biology, University of Illinois, Urbana, IL 61801 (United States); Chen Yi, E-mail: loganliu@illinois.edu [Department of Electrical and Computer Engineering, University of Illinois, Urbana, IL 61801 (United States)

    2011-09-07

    We have created a randomly distributed nanocone substrate on silicon coated with silver for surface-plasmon-enhanced fluorescence detection and 3D cell imaging. Optical characterization of the nanocone substrate showed it can support several plasmonic modes (in the 300-800 nm wavelength range) that can be coupled to a fluorophore on the surface of the substrate, which gives rise to the enhanced fluorescence. Spectral analysis suggests that a nanocone substrate can create more excitons and shorter lifetime in the model fluorophore Rhodamine 6G (R6G) due to plasmon resonance energy transfer from the nanocone substrate to the nearby fluorophore. We observed three-dimensional fluorescence enhancement on our substrate shown from the confocal fluorescence imaging of chinese hamster ovary (CHO) cells grown on the substrate. The fluorescence intensity from the fluorophores bound on the cell membrane was amplified more than 100-fold as compared to that on a glass substrate. We believe that strong scattering within the nanostructured area coupled with random scattering inside the cell resulted in the observed three-dimensional enhancement in fluorescence with higher photostability on the substrate surface.

  10. Fluorescent quantification of melanin.

    Fernandes, Bruno; Matamá, Teresa; Guimarães, Diana; Gomes, Andreia; Cavaco-Paulo, Artur

    2016-11-01

    Melanin quantification is reportedly performed by absorption spectroscopy, commonly at 405 nm. Here, we propose the implementation of fluorescence spectroscopy for melanin assessment. In a typical in vitro assay to assess melanin production in response to an external stimulus, absorption spectroscopy clearly overvalues melanin content. This method is also incapable of distinguishing non-melanotic/amelanotic control cells from those that are actually capable of performing melanogenesis. Therefore, fluorescence spectroscopy is the best method for melanin quantification as it proved to be highly specific and accurate, detecting even small variations in the synthesis of melanin. This method can also be applied to the quantification of melanin in more complex biological matrices like zebrafish embryos and human hair. © 2016 John Wiley & Sons A/S. Published by John Wiley & Sons Ltd.

  11. Fluorescent nanodiamond for biomedicine

    Milos Nesladek

    2014-01-01

    NV centers in diamond have gained strong interest as a novel tool for quantum information processing, quantum computing and quantum photonics. These applications are based on fluorescent and spin properties of NV-centres. However, in some conditions NV- can lose an electron and turn to NV0. The occupation of NV0 and NV- charge states depend on the position of their ground states with respect to the Fermi level and the mechanism of the charge transfer. Interestingly, that the charge switch has important implications on applications of fluorescent nanodiamond (fND) to nano-biology and nano-medicine. fND can be used for bio-marking and bio-tracking but also for the monitoring of targeted delivery to the cells. In this presentation we review the current state-of-the art for using fND particles for fluorescent bio imaging in cells and discuss the charge transfer and its luminescence stability by using ultra high sensitive spectroscopy methods to study the NV0 and NV- state occupation. (author)

  12. Stochastic resonance

    Wellens, Thomas; Shatokhin, Vyacheslav; Buchleitner, Andreas

    2004-01-01

    We are taught by conventional wisdom that the transmission and detection of signals is hindered by noise. However, during the last two decades, the paradigm of stochastic resonance (SR) proved this assertion wrong: indeed, addition of the appropriate amount of noise can boost a signal and hence facilitate its detection in a noisy environment. Due to its simplicity and robustness, SR has been implemented by mother nature on almost every scale, thus attracting interdisciplinary interest from physicists, geologists, engineers, biologists and medical doctors, who nowadays use it as an instrument for their specific purposes. At the present time, there exist a lot of diversified models of SR. Taking into account the progress achieved in both theoretical understanding and practical application of this phenomenon, we put the focus of the present review not on discussing in depth technical details of different models and approaches but rather on presenting a general and clear physical picture of SR on a pedagogical level. Particular emphasis will be given to the implementation of SR in generic quantum systems-an issue that has received limited attention in earlier review papers on the topic. The major part of our presentation relies on the two-state model of SR (or on simple variants thereof), which is general enough to exhibit the main features of SR and, in fact, covers many (if not most) of the examples of SR published so far. In order to highlight the diversity of the two-state model, we shall discuss several examples from such different fields as condensed matter, nonlinear and quantum optics and biophysics. Finally, we also discuss some situations that go beyond the generic SR scenario but are still characterized by a constructive role of noise

  13. State-dependent fluorescence of neutral atoms in optical potentials

    Martinez-Dorantes, M.; Alt, W.; Gallego, J.; Ghosh, S.; Ratschbacher, L.; Meschede, D.

    2018-02-01

    Recently we have demonstrated scalable, nondestructive, and high-fidelity detection of the internal state of 87Rb neutral atoms in optical dipole traps using state-dependent fluorescence imaging [M. Martinez-Dorantes, W. Alt, J. Gallego, S. Ghosh, L. Ratschbacher, Y. Völzke, and D. Meschede, Phys. Rev. Lett. 119, 180503 (2017), 10.1103/PhysRevLett.119.180503]. In this paper we provide experimental procedures and interpretations to overcome the detrimental effects of heating-induced trap losses and state leakage. We present models for the dynamics of optically trapped atoms during state-dependent fluorescence imaging and verify our results by comparing Monte Carlo simulations with experimental data. Our systematic study of dipole force fluctuations heating in optical traps during near-resonant illumination shows that off-resonant light is preferable for state detection in tightly confining optical potentials.

  14. Targeting pancreatic cancer with magneto-fluorescent theranostic gold nanoshells.

    Chen, Wenxue; Ayala-Orozco, Ciceron; Biswal, Nrusingh C; Perez-Torres, Carlos; Bartels, Marc; Bardhan, Rizia; Stinnet, Gary; Liu, Xian-De; Ji, Baoan; Deorukhkar, Amit; Brown, Lisa V; Guha, Sushovan; Pautler, Robia G; Krishnan, Sunil; Halas, Naomi J; Joshi, Amit

    2014-01-01

    We report a magneto-fluorescent theranostic nanocomplex targeted to neutrophil gelatinase-associated lipocalin (NGAL) for imaging and therapy of pancreatic cancer. Gold nanoshells resonant at 810 nm were encapsulated in silica epilayers doped with iron oxide and the near-infrared (NIR) dye indocyanine green, resulting in theranostic gold nanoshells (TGNS), which were subsequently conjugated with antibodies targeting NGAL in AsPC-1-derived xenografts in nude mice. Anti-NGAL-conjugated TGNS specifically targeted pancreatic cancer cells in vitro and in vivo providing contrast for both NIR fluorescence and T2-weighted MRI with higher tumor contrast than can be obtained using long-circulating, but nontargeted, PEGylated nanoparticles. The nanocomplexes also enabled highly specific cancer cell death via NIR photothermal therapy in vitro. TGNS with embedded NIR and magnetic resonance contrasts can be specifically targeted to pancreatic cancer cells with expression of early disease marker NGAL, and enable molecularly targeted imaging and photothermal therapy.

  15. Stem Cell-Based Neuroprotective and Neurorestorative Strategies

    Chia-Wei Hung

    2010-05-01

    Full Text Available Stem cells, a special subset of cells derived from embryo or adult tissues, are known to present the characteristics of self-renewal, multiple lineages of differentiation, high plastic capability, and long-term maintenance. Recent reports have further suggested that neural stem cells (NSCs derived from the adult hippocampal and subventricular regions possess the utilizing potential to develop the transplantation strategies and to screen the candidate agents for neurogenesis, neuroprotection, and neuroplasticity in neurodegenerative diseases. In this article, we review the roles of NSCs and other stem cells in neuroprotective and neurorestorative therapies for neurological and psychiatric diseases. We show the evidences that NSCs play the key roles involved in the pathogenesis of several neurodegenerative disorders, including depression, stroke and Parkinson’s disease. Moreover, the potential and possible utilities of induced pluripotent stem cells (iPS, reprogramming from adult fibroblasts with ectopic expression of four embryonic genes, are also reviewed and further discussed. An understanding of the biophysiology of stem cells could help us elucidate the pathogenicity and develop new treatments for neurodegenerative disorders. In contrast to cell transplantation therapies, the application of stem cells can further provide a platform for drug discovery and small molecular testing, including Chinese herbal medicines. In addition, the high-throughput stem cell-based systems can be used to elucidate the mechanisms of neuroprotective candidates in translation medical research for neurodegenerative diseases.

  16. Stem cell-based therapies for acute radiation syndrome

    Guha, Chandan

    2014-01-01

    Exposure to high doses of ionizing radiation in the event of accidental or intentional incident such as nuclear/radiological terrorism can lead to debilitating injuries to multiple organs resulting in death within days depending on the amount of radiation dose and the quality of radiation. Unfortunately, there is not a single FDA-licensed drug approved against acute radiation injury. The RadStem Center for Medical Countermeasures against Radiation (RadStem CMGR) program at Einstein is developing stem cell-based therapies to treat acute radiation syndrome (ARS). We have demonstrated that intravenous transplantation of bone marrow-derived and adipose-derived stromal cells, consisting of a mixture of mesenchymal, endothelial and myeloid progenitors can mitigate mice exposed to whole body irradiation of 12 Gy or whole abdominal irradiation of up to 20 Gy. We identified a variety of growth and differentiation factors that individually is unable to improve survival of animals exposed to lethal irradiation, but when administered sequentially mitigates radiation injury and improves survival. We termed this phenomenon as synthetic survival and describe a new paradigm whereby the 'synthetic survival' of irradiated tissues can be promoted by systemic administration of growth factors to amplify residual stem cell clonogens post-radiation exposure, followed by a differentiation factor that favors tissue stem cell differentiation. Synthetic survival can be applied to mitigate lethal radiation injury in multiple organs following radiation-induced hematopoeitic, gastrointestinal and pulmonary syndromes. (author)

  17. Potential of Stem Cell-Based Therapy for Ischemic Stroke

    Hany E. Marei

    2018-02-01

    Full Text Available Ischemic stroke is one of the major health problems worldwide. The only FDA approved anti-thrombotic drug for acute ischemic stroke is the tissue plasminogen activator. Several studies have been devoted to assessing the therapeutic potential of different types of stem cells such as neural stem cells (NSCs, mesenchymal stem cells, embryonic stem cells, and human induced pluripotent stem cell-derived NSCs as treatments for ischemic stroke. The results of these studies are intriguing but many of them have presented conflicting results. Additionally, the mechanism(s by which engrafted stem/progenitor cells exert their actions are to a large extent unknown. In this review, we will provide a synopsis of different preclinical and clinical studies related to the use of stem cell-based stroke therapy, and explore possible beneficial/detrimental outcomes associated with the use of different types of stem cells. Due to limited/short time window implemented in most of the recorded clinical trials about the use of stem cells as potential therapeutic intervention for stroke, further clinical trials evaluating the efficacy of the intervention in a longer time window after cellular engraftments are still needed.

  18. Microfluidic systems for stem cell-based neural tissue engineering.

    Karimi, Mahdi; Bahrami, Sajad; Mirshekari, Hamed; Basri, Seyed Masoud Moosavi; Nik, Amirala Bakhshian; Aref, Amir R; Akbari, Mohsen; Hamblin, Michael R

    2016-07-05

    Neural tissue engineering aims at developing novel approaches for the treatment of diseases of the nervous system, by providing a permissive environment for the growth and differentiation of neural cells. Three-dimensional (3D) cell culture systems provide a closer biomimetic environment, and promote better cell differentiation and improved cell function, than could be achieved by conventional two-dimensional (2D) culture systems. With the recent advances in the discovery and introduction of different types of stem cells for tissue engineering, microfluidic platforms have provided an improved microenvironment for the 3D-culture of stem cells. Microfluidic systems can provide more precise control over the spatiotemporal distribution of chemical and physical cues at the cellular level compared to traditional systems. Various microsystems have been designed and fabricated for the purpose of neural tissue engineering. Enhanced neural migration and differentiation, and monitoring of these processes, as well as understanding the behavior of stem cells and their microenvironment have been obtained through application of different microfluidic-based stem cell culture and tissue engineering techniques. As the technology advances it may be possible to construct a "brain-on-a-chip". In this review, we describe the basics of stem cells and tissue engineering as well as microfluidics-based tissue engineering approaches. We review recent testing of various microfluidic approaches for stem cell-based neural tissue engineering.

  19. Cryopreservation of GABAergic Neuronal Precursors for Cell-Based Therapy.

    Daniel Rodríguez-Martínez

    Full Text Available Cryopreservation protocols are essential for stem cells storage in order to apply them in the clinic. Here we describe a new standardized cryopreservation protocol for GABAergic neural precursors derived from the medial glanglionic eminence (MGE, a promising source of GABAergic neuronal progenitors for cell therapy against interneuron-related pathologies. We used 10% Me2SO as cryoprotectant and assessed the effects of cell culture amplification and cellular organization, as in toto explants, neurospheres, or individualized cells, on post-thaw cell viability and retrieval. We confirmed that in toto cryopreservation of MGE explants is an optimal preservation system to keep intact the interneuron precursor properties for cell transplantation, together with a high cell viability (>80% and yield (>70%. Post-thaw proliferation and self-renewal of the cryopreserved precursors were tested in vitro. In addition, their migration capacity, acquisition of mature neuronal morphology, and potency to differentiate into multiple interneuron subtypes were also confirmed in vivo after transplantation. The results show that the cryopreserved precursor features remained intact and were similar to those immediately transplanted after their dissection from the MGE. We hope this protocol will facilitate the generation of biobanks to obtain a permanent and reliable source of GABAergic precursors for clinical application in cell-based therapies against interneuronopathies.

  20. Microbial fuel cell based on electroactive sulfate-reducing biofilm

    Angelov, Anatoliy; Bratkova, Svetlana; Loukanov, Alexandre

    2013-01-01

    Highlights: ► Regulation and management of electricity generation by variation of residence time. ► Design of microbial fuel cell based on electroactive biofilm on zeolite. ► Engineering solution for removing of the obtained elemental sulfur. - abstract: A two chambered laboratory scale microbial fuel cell (MFC) has been developed, based on natural sulfate-reducing bacterium consortium in electroactive biofilm on zeolite. The MFC utilizes potassium ferricyanide in the cathode chamber as an electron acceptor that derives electrons from the obtained in anode chamber H 2 S. The molecular oxygen is finally used as a terminal electron acceptor at cathode compartment. The generated power density was 0.68 W m −2 with current density of 3.2 A m −2 at 150 Ω electrode resistivity. The hydrogen sulfide itself is produced by microbial dissimilative sulfate reduction process by utilizing various organic substrates. Finally, elemental sulfur was identified as the predominant final oxidation product in the anode chamber. It was removed from MFC through medium circulation and gathering in an external tank. This report reveals dependence relationship between the progress of general electrochemical parameters and bacterial sulfate-reduction rate. The presented MFC design can be used for simultaneous sulfate purification of mining drainage wastewater and generation of renewable electricity

  1. [Safety monitoring of cell-based medicinal products (CBMPs)].

    Funk, Markus B; Frech, Marion; Spranger, Robert; Keller-Stanislawski, Brigitte

    2015-11-01

    Cell-based medicinal products (CBMPs), a category of advanced-therapy medicinal products (ATMPs), are authorised for the European market by the European Commission by means of the centralized marketing authorisation. By conforming to the German Medicinal Products Act (Sec. 4b AMG), national authorisation can be granted by the Paul-Ehrlich-Institut in Germany exclusively for ATMPs not based on a routine manufacturing procedure. In both procedures, quality, efficacy, and safety are evaluated and the risk-benefit balance is assessed. For the centralised procedure, mainly controlled clinical trial data must be submitted, whereas the requirements for national procedures could be modified corresponding to the stage of development of the ATMP. After marketing authorization, the marketing authorization/license holder is obligated to report all serious adverse reactions to the competent authority and to provide periodic safety update reports. If necessary, post-authorization safety studies could be imposed. On the basis of these regulatory measures, the safety of advanced therapies can be monitored and improved.

  2. Stem Cell-Based Therapies for Polyglutamine Diseases.

    Mendonça, Liliana S; Onofre, Isabel; Miranda, Catarina Oliveira; Perfeito, Rita; Nóbrega, Clévio; de Almeida, Luís Pereira

    2018-01-01

    Polyglutamine (polyQ) diseases are a family of neurodegenerative disorders with very heterogeneous clinical presentations, although with common features such as progressive neuronal death. Thus, at the time of diagnosis patients might present an extensive and irreversible neuronal death demanding cell replacement or support provided by cell-based therapies. For this purpose stem cells, which include diverse populations ranging from embryonic stem cells (ESCs), to fetal stem cells, mesenchymal stromal cells (MSCs) or induced pluripotent stem cells (iPSCs) have remarkable potential to promote extensive brain regeneration and recovery in neurodegenerative disorders. This regenerative potential has been demonstrated in exciting pre and clinical assays. However, despite these promising results, several drawbacks are hampering their successful clinical implementation. Problems related to ethical issues, quality control of the cells used and the lack of reliable models for the efficacy assessment of human stem cells. In this chapter the main advantages and disadvantages of the available sources of stem cells as well as their efficacy and potential to improve disease outcomes are discussed.

  3. Coumarin amide derivatives as fluorescence chemosensors for cyanide anions

    Wu, Qianqian [School of Material Science and Engineering, Shandong Provincial Key Laboratory of Preparation and Measurement of Building Materials, University of Jinan, Jinan 250022, Shandong (China); Liu, Zhiqiang [State Key Laboratory of Crystal Materials, Shandong University, Jinan 250100, Shandong (China); Cao, Duxia, E-mail: duxiacao@ujn.edu.cn [School of Material Science and Engineering, Shandong Provincial Key Laboratory of Preparation and Measurement of Building Materials, University of Jinan, Jinan 250022, Shandong (China); Guan, Ruifang, E-mail: mse_guanrf@ujn.edu.cn [School of Material Science and Engineering, Shandong Provincial Key Laboratory of Preparation and Measurement of Building Materials, University of Jinan, Jinan 250022, Shandong (China); Wang, Kangnan; Shan, Yanyan; Xu, Yongxiao; Ma, Lin [School of Material Science and Engineering, Shandong Provincial Key Laboratory of Preparation and Measurement of Building Materials, University of Jinan, Jinan 250022, Shandong (China)

    2015-07-01

    Four coumarin amide derivatives with 4-methyl coumarin or pyrene as terminal group have been synthesized. Their photophysical properties and recognition properties for cyanide anions have been examined. The results indicate that the compounds can recognize cyanide anions with obvious absorption and fluorescence spectra change, at the same time, obvious color and fluorescence change can be observed by naked eye. The in situ hydrogen nuclear magnetic resonance spectra and photophysical properties change confirm that Michael additions between the chemosensors and cyanide anions take place at the 4-position of coumarin. - Highlights: • Four coumarin amide derivatives with 4-methyl coumarin or pyrene as terminal group were synthesized. • The compounds can recognize cyanide anions with obvious absorption and fluorescence spectra change. • Michael additions between the chemosensors and cyanide anions take place at the 4-position of coumarin.

  4. A molecular-sized optical logic circuit for digital modulation of a fluorescence signal

    Nishimura, Takahiro; Tsuchida, Karin; Ogura, Yusuke; Tanida, Jun

    2018-03-01

    Fluorescence measurement allows simultaneous detection of multiple molecular species by using spectrally distinct fluorescence probes. However, due to the broad spectra of fluorescence emission, the multiplicity of fluorescence measurement is generally limited. To overcome this limitation, we propose a method to digitally modulate fluorescence output signals with a molecular-sized optical logic circuit by using optical control of fluorescence resonance energy transfer (FRET). The circuit receives a set of optical inputs represented with different light wavelengths, and then it switches high and low fluorescence intensity from a reporting molecule according to the result of the logic operation. By using combinational optical inputs in readout of fluorescence signals, the number of biomolecular species that can be identified is increased. To implement the FRET-based circuits, we designed two types of basic elements, YES and NOT switches. An YES switch produces a high-level output intensity when receiving a designated light wavelength input and a low-level intensity without the light irradiation. A NOT switch operates inversely to the YES switch. In experiments, we investigated the operation of the YES and NOT switches that receive a 532-nm light input and modulate the fluorescence intensity of Alexa Fluor 488. The experimental result demonstrates that the switches can modulate fluorescence signals according to the optical input.

  5. Fluorescent microthermographic imaging

    Barton, D.L.

    1993-09-01

    In the early days of microelectronics, design rules and feature sizes were large enough that sub-micron spatial resolution was not needed. Infrared or IR thermal techniques were available that calculated the object`s temperature from infrared emission. There is a fundamental spatial resolution limitation dependent on the wavelengths of light being used in the image formation process. As the integrated circuit feature sizes began to shrink toward the one micron level, the limitations imposed on IR thermal systems became more pronounced. Something else was needed to overcome this limitation. Liquid crystals have been used with great success, but they lack the temperature measurement capabilities of other techniques. The fluorescent microthermographic imaging technique (FMI) was developed to meet this need. This technique offers better than 0.01{degrees}C temperature resolution and is diffraction limited to 0.3 {mu}m spatial resolution. While the temperature resolution is comparable to that available on IR systems, the spatial resolution is much better. The FMI technique provides better spatial resolution by using a temperature dependent fluorescent film that emits light at 612 nm instead of the 1.5 {mu}m to 12 {mu}m range used by IR techniques. This tutorial starts with a review of blackbody radiation physics, the process by which all heated objects emit radiation to their surroundings, in order to understand the sources of information that are available to characterize an object`s surface temperature. The processes used in infrared thermal imaging are then detailed to point out the limitations of the technique but also to contrast it with the FMI process. The FMI technique is then described in detail, starting with the fluorescent film physics and ending with a series of examples of past applications of FMI.

  6. Study of small-cell lung cancer cell-based sensor and its applications in chemotherapy effects rapid evaluation for anticancer drugs.

    Guohua, Hui; Hongyang, Lu; Zhiming, Jiang; Danhua, Zhu; Haifang, Wan

    2017-11-15

    Small cell lung cancer (SCLC) is a smoking-related cancer disease. Despite improvement in clinical survival, SCLC outcome remains extremely poor. Cisplatin (DDP) is the first-line chemotherapy drug for SCLC, but the choice of second-line chemotherapy drugs is not clear. In this paper, a SCLC cell-based sensor was proposed, and its applications in chemotherapy effects rapid evaluation for anticancer drugs were investigated. SCLC cell lines lung adenocarcinoma cell (LTEP-P) and DDP-resistant lung adenocarcinoma cell (LTEP-P/DDP-1.0) are cultured on carbon screen-printed electrode (CSPE) to fabricate integrated cell-based sensor. Several chemotherapy anticancer drugs, including cisplatin, ifosmamide, gemcitabine, paclitaxel, docetaxel, vinorelbine, etoposide, camptothecin, and topotecan, are selected as experimental chemicals. 3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) tests are conducted to evaluate chemotherapy drug effects on LTEP-P and LTEP-P/DDP-1.0 cell lines. Electrical cell-substrate impedance sensing (ECIS) responses to anti-tumor chemicals are measured and processed by double-layered cascaded stochastic resonance (DCSR). Cisplatin solutions in different concentrations measurement results demonstrate that LTEP-P cell-based sensor presents quantitative analysis abilities for cisplatin and topotecan. Cisplatin and its mixtures can also be discriminated. Results demonstrate that LTEP-P cell-based sensor sensitively evaluates chemotherapy drugs' apoptosis function to SCLC cells. LTEP-P/DDP-1.0 cell-based sensor responses demonstrate that gemcitabine, vinorelbine, and camptothecin are ideal second-line drugs for clinical post-cisplatin therapy than other drugs according to MTT test results. This work provides a novel way for SCLC second-line clinical chemotherapy drug screening. Copyright © 2017 Elsevier B.V. All rights reserved.

  7. Ethical and Safety Issues of Stem Cell-Based Therapy.

    Volarevic, Vladislav; Markovic, Bojana Simovic; Gazdic, Marina; Volarevic, Ana; Jovicic, Nemanja; Arsenijevic, Nebojsa; Armstrong, Lyle; Djonov, Valentin; Lako, Majlinda; Stojkovic, Miodrag

    2018-01-01

    Results obtained from completed and on-going clinical studies indicate huge therapeutic potential of stem cell-based therapy in the treatment of degenerative, autoimmune and genetic disorders. However, clinical application of stem cells raises numerous ethical and safety concerns. In this review, we provide an overview of the most important ethical issues in stem cell therapy, as a contribution to the controversial debate about their clinical usage in regenerative and transplantation medicine. We describe ethical challenges regarding human embryonic stem cell (hESC) research, emphasizing that ethical dilemma involving the destruction of a human embryo is a major factor that may have limited the development of hESC-based clinical therapies. With previous derivation of induced pluripotent stem cells (iPSCs) this problem has been overcome, however current perspectives regarding clinical translation of iPSCs still remain. Unlimited differentiation potential of iPSCs which can be used in human reproductive cloning, as a risk for generation of genetically engineered human embryos and human-animal chimeras, is major ethical issue, while undesired differentiation and malignant transformation are major safety issues. Although clinical application of mesenchymal stem cells (MSCs) has shown beneficial effects in the therapy of autoimmune and chronic inflammatory diseases, the ability to promote tumor growth and metastasis and overestimated therapeutic potential of MSCs still provide concerns for the field of regenerative medicine. This review offers stem cell scientists, clinicians and patient's useful information and could be used as a starting point for more in-depth analysis of ethical and safety issues related to clinical application of stem cells.

  8. High content cell-based assay for the inflammatory pathway

    Mukherjee, Abhishek; Song, Joon Myong

    2015-07-01

    Cellular inflammation is a non-specific immune response to tissue injury that takes place via cytokine network orchestration to maintain normal tissue homeostasis. However chronic inflammation that lasts for a longer period, plays the key role in human diseases like neurodegenerative disorders and cancer development. Understanding the cellular and molecular mechanisms underlying the inflammatory pathways may be effective in targeting and modulating their outcome. Tumor necrosis factor alpha (TNF-α) is a pro-inflammatory cytokine that effectively combines the pro-inflammatory features with the pro-apoptotic potential. Increased levels of TNF-α observed during acute and chronic inflammatory conditions are believed to induce adverse phenotypes like glucose intolerance and abnormal lipid profile. Natural products e. g., amygdalin, cinnamic acid, jasmonic acid and aspirin have proven efficacy in minimizing the TNF-α induced inflammation in vitro and in vivo. Cell lysis-free quantum dot (QDot) imaging is an emerging technique to identify the cellular mediators of a signaling cascade with a single assay in one run. In comparison to organic fluorophores, the inorganic QDots are bright, resistant to photobleaching and possess tunable optical properties that make them suitable for long term and multicolor imaging of various components in a cellular crosstalk. Hence we tested some components of the mitogen activated protein kinase (MAPK) pathway during TNF-α induced inflammation and the effects of aspirin in HepG2 cells by QDot multicolor imaging technique. Results demonstrated that aspirin showed significant protective effects against TNF-α induced cellular inflammation. The developed cell based assay paves the platform for the analysis of cellular components in a smooth and reliable way.

  9. A fluorescence scanning electron microscope

    Kanemaru, Takaaki; Hirata, Kazuho; Takasu, Shin-ichi; Isobe, Shin-ichiro; Mizuki, Keiji; Mataka, Shuntaro; Nakamura, Kei-ichiro

    2009-01-01

    Fluorescence techniques are widely used in biological research to examine molecular localization, while electron microscopy can provide unique ultrastructural information. To date, correlative images from both fluorescence and electron microscopy have been obtained separately using two different instruments, i.e. a fluorescence microscope (FM) and an electron microscope (EM). In the current study, a scanning electron microscope (SEM) (JEOL JXA8600 M) was combined with a fluorescence digital camera microscope unit and this hybrid instrument was named a fluorescence SEM (FL-SEM). In the labeling of FL-SEM samples, both Fluolid, which is an organic EL dye, and Alexa Fluor, were employed. We successfully demonstrated that the FL-SEM is a simple and practical tool for correlative fluorescence and electron microscopy.

  10. Resonance Raman and optical dephasing study of tricarbocyanine dyes

    Ashworth, SH; Kummrow, A; Lenz, K

    Fluorescence lineshape analysis based on resonance Raman spectra of the dye HITCI was used to determine the details and magnitude of the vibrational part of the line broadening function, Forced light scattering (FLS) was applied to measure optical dephasing of HITCI in ethylene glycol, pumping at

  11. Imaging in cell-based therapy for neurodegenerative diseases

    Kirik, Deniz; Breysse, Nathalie; Bjoerklund, Tomas; Besret, Laurent; Hantraye, Philippe

    2005-01-01

    Fetal cell transplantation for the treatment of Parkinson's and Huntington's diseases has been developed over the past two decades and is now in early clinical testing phase. Direct assessment of the graft's survival, integration into the host brain and impact on neuronal functions requires advanced in vivo neuroimaging techniques. Owing to its high sensitivity, positron emission tomography is today the most widely used tool to evaluate the viability and function of the transplanted tissue in the brain. Nuclear magnetic resonance techniques are opening new possibilities for imaging neurochemical events in the brain. The ultimate goal will be to use the combination of multiple imaging modalities for complete functional monitoring of the repair processes in the central nervous system. (orig.)

  12. Development of a fluorescent cryocooler

    Edwards, B.C.; Buchwald, M.I.; Epstein, R.I.; Gosnell, T.R.; Mungan, C.E.

    1995-01-01

    Recent work at Los Alamos National Laboratory has demonstrated the physical principles for a new type of solid-state cryocooler based on anti-Stokes fluorescence. Design studies indicate that a vibration-free, low-mass ''fluorescent cryocooler'' could operate for years with efficiencies and cooling powers comparable to current commercial systems. This paper presents concepts for a fluorescent cryocooler, design considerations and expected performance

  13. Fluorescence of ceramic color standards

    Koo, Annette; Clare, John F.; Nield, Kathryn M.; Deadman, Andrew; Usadi, Eric

    2010-01-01

    Fluorescence has been found in color standards available for use in calibration and verification of color measuring instruments. The fluorescence is excited at wavelengths below about 600 nm and emitted above 700 nm, within the response range of silicon photodiodes, but at the edge of the response of most photomultipliers and outside the range commonly scanned in commercial colorimeters. The degree of fluorescence on two of a set of 12 glossy ceramic tiles is enough to introduce significant error when those tiles have been calibrated in one mode of measurement and are used in another. We report the nature of the fluorescence and the implications for color measurement.

  14. Single-Cell Based Quantitative Assay of Chromosome Transmission Fidelity.

    Zhu, Jin; Heinecke, Dominic; Mulla, Wahid A; Bradford, William D; Rubinstein, Boris; Box, Andrew; Haug, Jeffrey S; Li, Rong

    2015-03-30

    Errors in mitosis are a primary cause of chromosome instability (CIN), generating aneuploid progeny cells. Whereas a variety of factors can influence CIN, under most conditions mitotic errors are rare events that have been difficult to measure accurately. Here we report a green fluorescent protein-based quantitative chromosome transmission fidelity (qCTF) assay in budding yeast that allows sensitive and quantitative detection of CIN and can be easily adapted to high-throughput analysis. Using the qCTF assay, we performed genome-wide quantitative profiling of genes that affect CIN in a dosage-dependent manner and identified genes that elevate CIN when either increased (icCIN) or decreased in copy number (dcCIN). Unexpectedly, qCTF screening also revealed genes whose change in copy number quantitatively suppress CIN, suggesting that the basal error rate of the wild-type genome is not minimized, but rather, may have evolved toward an optimal level that balances both stability and low-level karyotype variation for evolutionary adaptation. Copyright © 2015 Zhu et al.

  15. Concentration Dependence of Gold Nanoparticles for Fluorescence Enhancement

    Solomon, Joel; Wittmershaus, Bruce

    Noble metal nanoparticles possess a unique property known as surface plasmon resonance in which the conduction electrons oscillate due to incoming light, dramatically increasing their absorption and scattering of light. The oscillating electrons create a varying electric field that can affect nearby molecules. The fluorescence and photostability of fluorophores can be enhanced significantly when they are near plasmonic nanoparticles. This effect is called metal enhanced fluorescence (MEF). MEF from two fluorescence organic dyes, Lucifer Yellow CH and Riboflavin, was measured with different concentrations of 50-nm colloidal gold nanoparticles (Au-NP). The concentration range of Au-NP was varied from 2.5 to 250 pM. To maximize the interaction, the dyes were chosen so their emission spectra had considerable overlap with the absorption spectra of the Au-NP, which is common in MEF studies. If the dye molecules are too close to the surface of Au-NP, fluorescence quenching can occur instead of MEF. To try to observe this difference, silica-coated Au-NP were compared to citrate-based Au-NP; however, fluorescence quenching was observed with both Au-NP. This material is based upon work supported by the National Science Foundation under Grant Number NSF-ECCS-1306157.

  16. Efficient organic tandem solar cells based on small molecules

    Riede, Moritz; Widmer, Johannes; Timmreck, Ronny; Wynands, David; Leo, Karl [Institut fuer Angewandte Photophysik, Technische Universitaet Dresden, George-Baehr-Str. 1, 01069 Dresden (Germany); Uhrich, Christian; Schwartz, Gregor; Gnehr, Wolf-Michael; Hildebrandt, Dirk; Weiss, Andre; Pfeiffer, Martin [Heliatek GmbH, Treidlerstr. 3, 01139 Dresden (Germany); Hwang, Jaehyung; Sundarraj, Sudhakar; Erk, Peter [BASF SE, GVC/E-J542, 67056 Ludwigshafen (Germany)

    2011-08-23

    In this paper, two vacuum processed single heterojunction organic solar cells with complementary absorption are described and the construction and optimization of tandem solar cells based on the combination of these heterojunctions demonstrated. The red-absorbing heterojunction consists of C{sub 60} and a fluorinated zinc phthalocyanine derivative (F4-ZnPc) that leads to a 0.1-0.15 V higher open circuit voltage V{sub oc} than the commonly used ZnPc. The second heterojunction incorporates C{sub 60} and a dicyanovinyl-capped sexithiophene derivative (DCV6T) that mainly absorbs in the green. The combination of both heterojunctions into one tandem solar cell leads to an absorption over the whole visible range of the sun spectrum. Thickness variations of the transparent p-doped optical spacer between both subcells in the tandem solar cell is shown to lead to a significant change in short circuit current density j{sub sc} due to optical interference effects, whereas V{sub oc} and fill factor are hardly affected. The maximum efficiency {eta} of about 5.6% is found for a spacer thickness of 150-165 nm. Based on the optimized 165nm thick spacer, effects of intensity and angle of illumination, and temperature on a tandem device are investigated. Variations in illumination intensity lead to a linear change in j{sub sc} over three orders of magnitude and a nearly constant {eta} in the range of 30 to 310 mW cm{sup -2}. Despite the stacked heterojunctions, the performance of the tandem device is robust against different illumination angles: j{sub sc} and {eta} closely follow a cosine behavior between 0 and 70 . Investigations of the temperature behavior of the tandem device show an increase in {eta} of 0.016 percentage points per Kelvin between -20 C and 25 C followed by a plateau up to 50 C. Finally, further optimization of the tandem stack results in a certified {eta} of (6.07 {+-} 0.24)% on (1.9893 {+-} 0.0060)cm{sup 2} (Fraunhofer ISE), i.e., areas large enough to be of

  17. A dansyl-rhodamine ratiometric fluorescent probe for Hg2+ based on FRET mechanism.

    Xie, Puhui; Guo, Fengqi; Wang, Lingyu; Yang, Sen; Yao, Denghui; Yang, Guoyu

    2015-03-01

    Based on resonance energy transfer (FRET) from dansyl to rhodamine 101, a new fluorescent probe (compound 1) containing rhodamine 101 and a dansyl unit was synthesized for detecting Hg(2+) through ratiometric sensing in DMSO aqueous solutions. This probe shows a fast, reversible and selective response toward Hg(2+) in a wide pH range. Hg(2+) induced ring-opening reactions of the spirolactam rhodamine moiety of 1, leading to the formation of fluorescent derivatives that can serve as the FRET acceptors. Very large stokes shift (220 nm) was observed in this case. About 97-fold increase in fluorescence intensity ratio was observed upon its binding with Hg(2+).

  18. Calibrating the photo-thermal response of magneto-fluorescent gold nanoshells.

    Biswal, Nrusingh C; Ayala-Orzoco, Ciceron; Halas, Naomi J; Joshi, Amit

    2011-01-01

    We report the photothermal response and Near Infrared (NIR) imaging sensitivities of magneto-fluorescent silica core gold nanocomplexes designed for molecular image guided thermal therapy of cancer. Approximately 160 nm Silica core gold nanoshells were designed to provide NIR fluorescent and Magnetic Resonance (MR) contrast by incorporating FDA approved dye indocyanine green (ICG) and iron-oxide within an outer silica epilayer. The imaging and therapeutic sensitivity, and the stability of fluorescence contrast for 12 microliters of suspension (containing approximately 7.9 × 10(8) or 1.3 femtoMole nanoshells) buried at depths of 2-8 mm in tissue mimicking scattering media is reported.

  19. Fluorescing macerals from wood precursors

    Stout, S A; Bensley, D F

    1987-01-01

    A preliminary investigation into the origin of wood-derived macerals has established the existence of autofluorescent maceral precursors in the secondary xylem of swamp-inhabiting plant species. The optical character and fluorescent properties of microtomed thin-sections of modern woods from the Florida Everglades and Okefenokee Swamp, Georgia are compared to the character and properties of their peatified equivalents from various Everglades and Okefenokee peat horizons and their lignitic equivalents from the Brandon lignite of Vermont and the Trail Ridge lignitic peat from northern Florida. The inherent fluorescence of woody cell walls is believed to be caused by lignin though other cell wall components may contribute. The fluorescence spectra for several wood and cell types had a ..gamma../sub m//sub a//sub x/ of 452 nm and Q value of 0.00. The color as observed in blue light and the spectral geometry as measured in UV light of peatified and lignitic woody cell walls (potential textinites) may change progressively during early coalification. Cell wall-derived maceral material is shown to maintain its fluorescing properties after being converted to a structureless material, perhaps a corpohuminite or humodetrinite precursor. Fluorescing xylem cell contents, such as condensed tannins or essential oils, can maintain the fluorescent character through early coalification. Xylem cell walls and xylem cell contents are shown to provide fluorescing progenitor materials which would not require subsequent infusion with 'lipid' materials to account for their fluorescence as phytoclast material or as macerals in coal. 35 references.

  20. Assessing Photosynthesis by Fluorescence Imaging

    Saura, Pedro; Quiles, Maria Jose

    2011-01-01

    This practical paper describes a novel fluorescence imaging experiment to study the three processes of photochemistry, fluorescence and thermal energy dissipation, which compete during the dissipation of excitation energy in photosynthesis. The technique represents a non-invasive tool for revealing and understanding the spatial heterogeneity in…

  1. Crossing simple resonances

    Collins, T.

    1985-08-01

    A simple criterion governs the beam distortion and/or loss of protons on a fast resonance crossing. Results from numerical integrations are illustrated for simple sextupole, octupole, and 10-pole resonances

  2. Acoustic Fano resonators

    Amin, Muhammad; Farhat, Mohamed; Bagci, Hakan

    2014-01-01

    The resonances with asymmetric Fano line-shapes were originally discovered in the context of quantum mechanics (U. Fano, Phys. Rev., 124, 1866-1878, 1961). Quantum Fano resonances were generated from destructive interference of a discrete state

  3. Neutron resonance averaging

    Chrien, R.E.

    1986-10-01

    The principles of resonance averaging as applied to neutron capture reactions are described. Several illustrations of resonance averaging to problems of nuclear structure and the distribution of radiative strength in nuclei are provided. 30 refs., 12 figs

  4. Crossing simple resonances

    Collins, T.

    1985-08-01

    A simple criterion governs the beam distortion and/or loss of protons on a fast resonance crossing. Results from numerical integrations are illustrated for simple sextupole, octupole, and 10-pole resonances.

  5. Time-resolved fluorescence spectroscopy

    Gustavsson, Thomas; Mialocq, Jean-Claude

    2007-01-01

    This article addresses the evolution in time of light emitted by a molecular system after a brief photo-excitation. The authors first describe fluorescence from a photo-physical point of view and discuss the characterization of the excited state. Then, they explain some basic notions related to fluorescence characterization (lifetime and decays, quantum efficiency, so on). They present the different experimental methods and techniques currently used to study time-resolved fluorescence. They discuss basic notions of time resolution and spectral reconstruction. They briefly present some conventional methods: intensified Ccd cameras, photo-multipliers and photodiodes associated with a fast oscilloscope, and phase modulation. Other methods and techniques are more precisely presented: time-correlated single photon counting (principle, examples, and fluorescence lifetime imagery), streak camera (principle, examples), and optical methods like the Kerr optical effect (principle and examples) and fluorescence up-conversion (principle and theoretical considerations, examples of application)

  6. Pediatric magnetic resonance imaging

    Cohen, M.D.

    1986-01-01

    This book defines the current clinical potential of magnetic resonance imaging and focuses on direct clinical work with pediatric patients. A section dealing with the physics of magnetic resonance imaging provides an introduction to enable clinicians to utilize the machine and interpret the images. Magnetic resonance imaging is presented as an appropriate imaging modality for pediatric patients utilizing no radiation

  7. Resonant thermonuclear reaction rate

    Haubold, H.J.; Mathai, A.M.

    1986-01-01

    Basic physical principles for the resonant and nonresonant thermonuclear reaction rates are applied to find their standard representations for nuclear astrophysics. Closed-form representations for the resonant reaction rate are derived in terms of Meijer's G-function. Analytic representations of the resonant and nonresonant nuclear reaction rates are compared and the appearance of Meijer's G-function is discussed in physical terms

  8. Single Molecule Spectroscopy of Fluorescent Proteins

    Blum, Christian; Subramaniam, Vinod

    2009-01-01

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

  9. Fluorescent multiplex cell flow systems and methods

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

    2017-01-01

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

  10. Quantum mechanical resonances

    Cisneros S, A.; McIntosh, H.V.

    1982-01-01

    A discussion of the nature of quantum mechanical resonances is presented from the point of view of the spectral theory of operators. In the case of Bohr-Feshbach resonances, graphs are presented to illustrate the theory showing the decay of a doubly excited metastable state and the excitation of the resonance by an incident particle with proper energy. A characterization of resonances is given as well as a procedure to determine widths using the spectral density function. A sufficient condition is given for the validity of the Breit-Wigner formula for Bohr-Feshbach resonances. (author)

  11. Generally Applicable Transformation Protocols for Fluorescent Nanodiamond Internalization into Cells.

    Hemelaar, Simon R; van der Laan, Kiran J; Hinterding, Sophie R; Koot, Manon V; Ellermann, Else; Perona-Martinez, Felipe P; Roig, David; Hommelet, Severin; Novarina, Daniele; Takahashi, Hiroki; Chang, Michael; Schirhagl, Romana

    2017-07-19

    Fluorescent nanodiamonds (FNDs) are promising nanoprobes, owing to their stable and magnetosensitive fluorescence. Therefore they can probe properties as magnetic resonances, pressure, temperature or strain. The unprecedented sensitivity of diamond defects can detect the faint magnetic resonance of a single electron or even a few nuclear spins. However, these sensitivities are only achieved if the diamond probe is close to the molecules that need to be detected. In order to utilize its full potential for biological applications, the diamond particle has to enter the cell. Some model systems, like HeLa cells, readily ingest particles. However, most cells do not show this behavior. In this article we show for the first time generally applicable methods, which are able to transport fluorescent nanodiamonds into cells with a thick cell wall. Yeast cells, in particular Saccharomyces cerevisiae, are a favored model organism to study intracellular processes including aging on a cellular level. In order to introduce FNDs in these cells, we evaluated electrical transformation and conditions of chemical permeabilization for uptake efficiency and viability. 5% DMSO (dimethyl sulfoxide) in combination with optimized chemical transformation mix leads to high uptake efficiency in combination with low impact on cell biology. We have evaluated all steps in the procedure.

  12. Fluorescent standards for photodynamic therapy

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

    2016-08-01

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

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

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

    2015-01-01

    the vps10p domain receptor sortilin and the neurotrophin receptor p75(NTR). However, proNGF-induced receptor complex formation has been difficult to directly assess other than by western blotting. We here describe a fluorescence resonance energy transfer (FRET) based fluorescence plate reader assay...

  14. Optically trapped atomic resonant devices for narrow linewidth spectral imaging

    Qian, Lipeng

    This thesis focuses on the development of atomic resonant devices for spectroscopic applications. The primary emphasis is on the imaging properties of optically thick atomic resonant fluorescent filters and their applications. In addition, this thesis presents a new concept for producing very narrow linewidth light as from an atomic vapor lamp pumped by a nanosecond pulse system. This research was motivated by application for missile warning system, and presents an innovative approach to a wide angle, ultra narrow linewidth imaging filter using a potassium vapor cell. The approach is to image onto and collect the fluorescent photons emitted from the surface of an optically thick potassium vapor cell, generating a 2 GHz pass-band imaging filter. This linewidth is narrow enough to fall within a Fraunhefer dark zone in the solar spectrum, thus make the detection solar blind. Experiments are conducted to measure the absorption line shape of the potassium resonant filter, the quantum efficiency of the fluorescent behavior, and the resolution of the fluorescent image. Fluorescent images with different spatial frequency components are analyzed by using a discrete Fourier transform, and the imaging capability of the fluorescent filter is described by its Modulation Transfer Function. For the detection of radiation that is spectrally broader than the linewidth of the potassium imaging filter, the fluorescent image is seen to be blurred by diffuse fluorescence from the slightly off resonant photons. To correct this, an ultra-thin potassium imaging filter is developed and characterized. The imaging property of the ultra-thin potassium imaging cell is tested with a potassium seeded flame, yielding a resolution image of ˜ 20 lines per mm. The physics behind the atomic resonant fluorescent filter is radiation trapping. The diffusion process of the resonant photons trapped in the atomic vapor is theoretically described in this thesis. A Monte Carlo method is used to simulate the

  15. Fluorescence molecular tomography in the presence of background fluorescence

    Soubret, Antoine; Ntziachristos, Vasilis

    2006-01-01

    Fluorescence molecular tomography is an emerging imaging technique that resolves the bio-distribution of engineered fluorescent probes developed for in vivo reporting of specific cellular and sub-cellular targets. The method can detect fluorochromes in picomole amounts or less, imaged through entire animals, but the detection sensitivity and imaging performance drop in the presence of background, non-specific fluorescence. In this study, we carried out a theoretical and an experimental investigation on the effect of background fluorescence on the measured signal and on the tomographic reconstruction. We further examined the performance of three subtraction methods based on physical models of photon propagation, using experimental data on phantoms and small animals. We show that the data pre-processing with subtraction schemes can improve image quality and quantification when non-specific background florescence is present

  16. Alloy catalysts for fuel cell-based alcohol sensors

    Ghavidel, Mohammadreza Zamanzad

    Direct ethanol fuel cells (DEFCs) are attractive from both economic and environmental standpoints for generating renewable energy and powering vehicles and portable electronic devices. There is a great interest recently in developing DEFC systems. The cost and performance of the DEFCs are mainly controlled by the Pt-base catalysts used at each electrode. In addition to energy conversion, DEFC technology is commonly employed in the fuel-cell based breath alcohol sensors (BrAS). BrAS is a device commonly used to measure blood alcohol concentration (BAC) and enforce drinking and driving laws. The BrAS is non-invasive and has a fast respond time. However, one of the most important drawback of the commercially available BrAS is the very high loading of Pt employed. One well-known and cost effective method to reduce the Pt loading is developing Pt-alloy catalysts. Recent studies have shown that Pt-transition metal alloy catalysts enhanced the electroactivity while decreasing the required loadings of the Pt catalysts. In this thesis, carbon supported Pt-Mn and Pt-Cu electrocatalysts were synthesized by different methods and the effects of heat treatment and structural modification on the ethanol oxidation reaction (EOR) activity, oxygen reduction reaction (ORR) activity and durability of these samples were thoroughly studied. Finally, the selected Pt-Mn and Pt-Cu samples with the highest EOR activity were examined in a prototype BrAS system and compared to the Pt/C and Pt 3Sn/C commercial electrocatalysts. Studies on the Pt-Mn catalysts produced with and without additives indicate that adding trisodium citrate (SC) to the impregnation solution improved the particle dispersion, decreased particle sizes and reduced the time required for heat treatment. Further studies show that the optimum weight ratio of SC to the metal loading in the impregnation solution was 2:1 and optimum results achieved at pH lower than 4. In addition, powder X-ray diffraction (XRD) analyses indicate

  17. Resonance energy transfer: Dye to metal nanoparticles

    Wari, M. N.; Pujar, G. H.; Inamdar, S. R., E-mail: him-lax3@yahoo.com [Laser Spectroscopy Programme, Department of Physics, Karnatak University, Dharwad-580003 (India)

    2015-06-24

    In the present study, surface energy transfer (SET) from Coumarin 540A (C540 A) to Gold nanoparticle (Au) is demonstrated. The observed results show pronounced effect on the photoluminescence intensity and shortening of the lifetime of Coumarin 540A upon interaction with the spherical gold nanoparticle, also there are measured effects on radiative rate of the dye. Experimental results are analyzed with fluorescence resonance energy transfer (FRET) and SET theories. The results obtained from distance-dependent quenching provide experimental evidence that the efficiency curve slope and distance of quenching is best modeled by surface energy transfer process.

  18. Thousand-fold enhancement of single-molecule fluorescence near a single gold nanorod

    Yuan, H.; Khatua, S.; Zijlstra, P.; Yorulmaz, M.; Orrit, M.

    2013-01-01

    Single molecules: Large enhancements of single-molecule fluorescence up to 1100 times by using synthesized gold nanorods are reported (see picture). This high enhancement is achieved by selecting a dye with its adsorption and emission close to the surface plasmon resonance of the gold nanorods

  19. Microstrip resonators for electron paramagnetic resonance experiments

    Torrezan, A. C.; Mayer Alegre, T. P.; Medeiros-Ribeiro, G.

    2009-07-01

    In this article we evaluate the performance of an electron paramagnetic resonance (EPR) setup using a microstrip resonator (MR). The design and characterization of the resonator are described and parameters of importance to EPR and spin manipulation are examined, including cavity quality factor, filling factor, and microwave magnetic field in the sample region. Simulated microwave electric and magnetic field distributions in the resonator are also presented and compared with qualitative measurements of the field distribution obtained by a perturbation technique. Based on EPR experiments carried out with a standard marker at room temperature and a MR resonating at 8.17 GHz, the minimum detectable number of spins was found to be 5×1010 spins/GHz1/2 despite the low MR unloaded quality factor Q0=60. The functionality of the EPR setup was further evaluated at low temperature, where the spin resonance of Cr dopants present in a GaAs wafer was detected at 2.3 K. The design and characterization of a more versatile MR targeting an improved EPR sensitivity and featuring an integrated biasing circuit for the study of samples that require an electrical contact are also discussed.

  20. Microstrip resonators for electron paramagnetic resonance experiments.

    Torrezan, A C; Mayer Alegre, T P; Medeiros-Ribeiro, G

    2009-07-01

    In this article we evaluate the performance of an electron paramagnetic resonance (EPR) setup using a microstrip resonator (MR). The design and characterization of the resonator are described and parameters of importance to EPR and spin manipulation are examined, including cavity quality factor, filling factor, and microwave magnetic field in the sample region. Simulated microwave electric and magnetic field distributions in the resonator are also presented and compared with qualitative measurements of the field distribution obtained by a perturbation technique. Based on EPR experiments carried out with a standard marker at room temperature and a MR resonating at 8.17 GHz, the minimum detectable number of spins was found to be 5 x 10(10) spins/GHz(1/2) despite the low MR unloaded quality factor Q0=60. The functionality of the EPR setup was further evaluated at low temperature, where the spin resonance of Cr dopants present in a GaAs wafer was detected at 2.3 K. The design and characterization of a more versatile MR targeting an improved EPR sensitivity and featuring an integrated biasing circuit for the study of samples that require an electrical contact are also discussed.

  1. A Starting Point for Fluorescence-Based Single-Molecule Measurements in Biomolecular Research

    Alexander Gust

    2014-09-01

    Full Text Available Single-molecule fluorescence techniques are ideally suited to provide information about the structure-function-dynamics relationship of a biomolecule as static and dynamic heterogeneity can be easily detected. However, what type of single-molecule fluorescence technique is suited for which kind of biological question and what are the obstacles on the way to a successful single-molecule microscopy experiment? In this review, we provide practical insights into fluorescence-based single-molecule experiments aiming for scientists who wish to take their experiments to the single-molecule level. We especially focus on fluorescence resonance energy transfer (FRET experiments as these are a widely employed tool for the investigation of biomolecular mechanisms. We will guide the reader through the most critical steps that determine the success and quality of diffusion-based confocal and immobilization-based total internal reflection fluorescence microscopy. We discuss the specific chemical and photophysical requirements that make fluorescent dyes suitable for single-molecule fluorescence experiments. Most importantly, we review recently emerged photoprotection systems as well as passivation and immobilization strategies that enable the observation of fluorescently labeled molecules under biocompatible conditions. Moreover, we discuss how the optical single-molecule toolkit has been extended in recent years to capture the physiological complexity of a cell making it even more relevant for biological research.

  2. Time-resolved UV-excited microarray reader for fluorescence energy transfer (FRET) measurements

    Orellana, Adelina; Hokkanen, Ari P.; Pastinen, Tomi; Takkinen, Kristina; Soderlund, Hans

    2001-05-01

    Analytical systems based on immunochemistry are largely used in medical diagnostics and in biotechnology. There is a significant pressure to develop the present assay formats to become easier to use, faster, and less reagent consuming. Further developments towards high density array--like multianalyte measurement systems would be valuable. To this aim we have studied the applicability of fluorescence resonance energy transfer and time-resolved fluorescence resonance energy transfer in immunoassays on microspots and in microwells. We have used engineered recombinant antibodies detecting the pentameric protein CRP as a model analyte system, and tested different assay formats. We describe also the construction of a time-resolved scanning epifluorometer with which we could measure the FRET interaction between the slow fluorescence decay from europium chelates and its energy transfer to the rapidly decaying fluorophore Cy5.

  3. Chromophore Structure of Photochromic Fluorescent Protein Dronpa: Acid-Base Equilibrium of Two Cis Configurations.

    Higashino, Asuka; Mizuno, Misao; Mizutani, Yasuhisa

    2016-04-07

    Dronpa is a novel photochromic fluorescent protein that exhibits fast response to light. The present article is the first report of the resonance and preresonance Raman spectra of Dronpa. We used the intensity and frequency of Raman bands to determine the structure of the Dronpa chromophore in two thermally stable photochromic states. The acid-base equilibrium in one photochromic state was observed by spectroscopic pH titration. The Raman spectra revealed that the chromophore in this state shows a protonation/deprotonation transition with a pKa of 5.2 ± 0.3 and maintains the cis configuration. The observed resonance Raman bands showed that the other photochromic state of the chromophore is in a trans configuration. The results demonstrate that Raman bands selectively enhanced for the chromophore yield valuable information on the molecular structure of the chromophore in photochromic fluorescent proteins after careful elimination of the fluorescence background.

  4. Instructive for disposal of fluorescent

    Salazar Vargas, Gerlin

    2014-01-01

    An instructive is established for the management system of waste fluorescent lamps, ensuring the storage, collection, transportation, and final disposal. The lamp is changed by an official of the Seccion de Matenimiento Construccion of the Oficina de Servicios Generales or is produced with the support of an official of the unit. The fluorescent should be deposited in stock of materials of the building maintenance section or unit specified with the help of a staff and in appropriate conditions. The fluorescent lamp is transported according to the guidelines in the manual. A responsible company is contracted by la Vicerrectoria de Administracion of the Universidad de Costa Rica dedicated to the transport and proper handling of fluorescent lamps [es

  5. ANTAGONISTIC POTENTIAL OF FLUORESCENT Pseudomonas ...

    Prof. Adipala Ekwamu

    GROWTH OF TOMATO CHALLENGED WITH PHTOPATHOGENS ... This study focused on the antagonistic potential of fluorescent Pseudomonas in vitro, and its inoculation effect on growth .... the 5 days old culture in starch agar with Lugol's.

  6. Low Sensitivity of T-Cell Based Detection of Tuberculosis among ...

    Low Sensitivity of T-Cell Based Detection of Tuberculosis among HIV Co-Infected Tanzanian In-Patients. ... with and without HIV infection. Design: Cross-sectional study. ... like Tanzania. Larger studies in resource-poor settings are required.

  7. X-ray fluorescence holography

    Hayashi, K; Takahashi, Y

    2003-01-01

    X-ray fluorescence holography (XFH) is a new structural analysis method of determining a 3D atomic arrangement around fluorescing atoms. We developed an XFH apparatus using advanced X-ray techniques and succeeded in obtaining high-quality hologram data. Furthermore, we introduced applications to the structural analysis of a thin film and the environment around dopants and, discussed the quantitative analysis of local lattice distortion. (author)

  8. Cell Based GIS as Cellular Automata for Disaster Spreading Predictions and Required Data Systems

    Kohei Arai

    2013-03-01

    Full Text Available A method for prediction and simulation based on the Cell Based Geographic Information System(GIS as Cellular Automata (CA is proposed together with required data systems, in particular metasearch engine usage in an unified way. It is confirmed that the proposed cell based GIS as CA has flexible usage of the attribute information that is attached to the cell in concert with location information and does work for disaster spreading simulation and prediction.

  9. Fluorescent Nanodiamonds in Biomedical Applications.

    Mitura, Katarzyna Anna; Włodarczyk, Elżbieta

    2018-04-18

    Nanoparticles have an extended surface and a large surface area, which is the ratio of the size of the surfacearea to the volume. A functionalized surface can give rise to more modifications and therefore allows this nanomaterial to have new properties. Fluorescent molecules contain fluorophore, which is capable of being excited via the absorption of light energy at a specific wavelength and subsequently emitting radiation energy of a longer wavelength. A chemically modified surface of nanodiamond (ND; by carboxylation) demonstrated biocompatibility with DNA, cytochrome C, and antigens. In turn, fluorescent nanodiamonds (FNDs) belong to a group of new nanomaterials. Their surface can be modified by joining functional groups such as carboxyl, hydroxyl, or amino, after which they can be employed as a fluorescence agent. Their fluorescent properties result from defects in the crystal lattice. FNDs reach dimensions of 4-100 nm, have attributes such as photostability, long fluorescence lifetimes (10 ns), and fluorescence emission between 600 and 700 nm. They are also nontoxic, chemically inert, biocompatible, and environmentally harmless. The main purpose of this article was to present the medical applications of various types of modified NDs.

  10. Fluorescence detection of esophageal neoplasia

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

    2008-06-01

    White-light endoscopy is well-established and wide used modality. However, despite the many technological advances that have been occurred, conventional endoscopy is suboptimal and usually detects advanced stage lesions. The limitations of standard endoscopy initiate development of spectroscopic techniques, additional to standard endoscopic equipment. One of the most sensitive approaches is fluorescence spectroscopy of gastrointestinal mucosa for neoplasia detection. In the recent study delta-aminolevulinic acid/Protoporphyrin IX (5-ALA/PpIX) is used as fluorescent marker for dysplasia and tumor detection in esophagus. The 5-ALA is administered per os six hours before measurements at dose 20 mg/kg weight. Excitation source has max of emission at 405 nm and light is delivered by the standard light guide of the endoscopic equipment. Through endoscopic instrumental channel a fiber is applied to return information about fluorescence to microspectrometer. Spectral features observed during endoscopic investigations could be distinct as the next regions: 450-630 nm region, where tissue autofluorescence is observed; 630-710 nm region, where fluorescence of PpIX is clearly pronounced; 530-580 nm region, where minima in the autofluorescence signal are observed, related to reabsorption of blood. The lack of fluorescence peaks in the red spectral area for normal mucosa is an indication for selective accumulation of 5-ALA/PpIX only in abnormal sites Very good correlation between fluorescence signals and histology examination of the lesions investigated is achieved.

  11. Spectroscopic Analysis of Red Fluorescent Proteins and Development of a Microfluidic Cell Sorter for the Generation of Improved Variants

    Lubbeck, Jennifer L.

    The discovery of the green fluorescent protein (GFP) launched the development of a wide variety of fluorescent protein (FP) mutants whose spectral and photophysical diversity revolutionized in vivo imaging. The excitation and emission spectra of red fluorescent proteins (RFPs), in particular, have been ideally tuned to a window optically favorable for in vivo work. However, their quantum yields, photostabilities and fluorescence intermittency properties require improvement if they are to be broadly employed for low-copy or single-molecule measurements. Attempts to engineer improved RFPs often result in optimization of one photophysical property at the expense of others. We developed a microfluidic-based cytometer for screening HeLa cell-based genetic RFP-libraries simultaneously on the basis of fluorescence lifetime (a proxy for quantum yield), photostability, and brightness. Ten 532 nm excitation beams interrogate each cell in flow. The first is electro-optically modulated (30 MHz) to enable lifetime measurement with phase fluorimetry. The remaining beams act as a pulse sequence for isolating the irreversible photobleaching time constant. Optical-force switching is employed to sort cells based on any combination of the photophysical parameters. Screening with this instrument enables identification of regions of the structure that synergistically affect quantum yield and photostability and the sorting capability provides a new tool for accelerating the development of next generation RFPs.

  12. Atlas of neutron resonances

    Mughabghab, Said

    2018-01-01

    Atlas of Neutron Resonances: Resonance Properties and Thermal Cross Sections Z= 1-60, Sixth Edition, contains an extensive list of detailed individual neutron resonance parameters for Z=1-60, as well as thermal cross sections, capture resonance integrals, average resonance parameters and a short survey of the physics of thermal and resonance neutrons. The long introduction contains: nuclear physics formulas aimed at neutron physicists; topics of special interest such as valence neutron capture, nuclear level density parameters, and s-, p-, and d-wave neutron strength functions; and various comparisons of measured quantities with the predictions of nuclear models, such as the optical model. As in the last edition, additional features have been added to appeal to a wider spectrum of users. These include: spin-dependent scattering lengths that are of interest to solid-state physicists, nuclear physicists and neutron evaluators; calculated and measured Maxwellian average 5-keV and 30-keV capture cross sections o...

  13. FLUORESCENCE DIAGNOSIS FOR RECURRENT BLADDER CANCER

    R. V. Ulyanov

    2017-01-01

    Full Text Available The clinical case of successful use of local fluorescence spectroscopy combined with fluorescence imaging during cystoscopy for diagnosis of recurrent bladder cancer is represented in the article. Histological study of fluorescent foci confirmed tumor growth (urothelial carcinoma in all areas with high levels of diagnostic parameter. In the fluorescent focus with low diagnostic parameter inflammation was detected.

  14. Magnetic resonance imaging apparatus

    Ehnholm, G.J.

    1991-01-01

    This patent describes an electron spin resonance enhanced magnetic resonance (MR) imaging (ESREMRI) apparatus able to generate a primary magnetic field during periods of nuclear spin transition excitation and magnetic resonance signal detection. This allows the generation of ESREMRI images of a subject. A primary magnetic field of a second and higher value generated during periods of nuclear spin transition excitation and magnetic resonance signal detection can be used to generate conventional MR images of a subject. The ESREMRI and native MR images so generated may be combined, (or superimposed). (author)

  15. Electron paramagnetic resonance

    Al'tshuler, S A

    2013-01-01

    Electron Paramagnetic Resonance is a comprehensive text on the field of electron paramagnetic resonance, covering both the theoretical background and the results of experiment. This book is composed of eight chapters that cover theoretical materials and experimental data on ionic crystals, since these are the materials that have been most extensively studied by the methods of paramagnetic resonance. The opening chapters provide an introduction to the basic principles of electron paramagnetic resonance and the methods of its measurement. The next chapters are devoted to the theory of spectra an

  16. Ramifide resonators for cyclotrons

    Smirnov, Yu.V.

    2000-01-01

    The resonators with the conductors ramified form for cyclotrons are systematized and separated into the self-contained class - the ramified resonators for cyclotrons (Carr). The ramified resonators are compared with the quarter-wave and half-wave nonramified resonators, accomplished from the transmitting lines fragments. The CRR are classified into two types: ones with the additional structural element, switched in parallel and in series. The CRR may include several additional structural elements. The CRR calculations may be concluded by analytical methods - the method of matrix calculation or the method of telegraph equations and numerical methods - by means of the ISFEL3D, MAFIA and other programs [ru

  17. Mechanotransduction in Endothelial Cells Studied with Fluorescence Imaging

    Chien Shu [Departments of Bioengineering and Medicine and Institute of Engineering in Medicine, University of California, San Diego, La Jolla, California 92093-0427 (United States)

    2011-01-01

    Mechanotransduction involves the conversion of mechanical stimuli to intracellular signaling to modulate gene and protein expressions and hence cellular functions in endothelial cells, thus playing importance roles in the regulation of homeostasis in health and disease. The aim of this paper is to investigate the dynamics of mechanotransduction in endothelial cells by the use of fluorescent resonance energy transfer (FRET) to study the temporal and spatial activation of Src kinase and focal adhesion kinase, both of which play critical roles in many cellular processes. The results have contributed to the elucidation of the roles of these two important signaling molecules and their interactions in mediating mechanotransduction.

  18. Deep UV Native Fluorescence Imaging of Antarctic Cryptoendolithic Communities

    Storrie-Lombardi, M. C.; Douglas, S.; Sun, H.; McDonald, G. D.; Bhartia, R.; Nealson, K. H.; Hug, W. F.

    2001-01-01

    An interdisciplinary team at the Jet Propulsion Laboratory Center for Life Detection has embarked on a project to provide in situ chemical and morphological characterization of Antarctic cryptoendolithic microbial communities. We present here in situ deep ultraviolet (UV) native fluorescence and environmental scanning electron microscopy images transiting 8.5 mm into a sandstone sample from the Antarctic Dry Valleys. The deep ultraviolet imaging system employs 224.3, 248.6, and 325 nm lasers to elicit differential fluorescence and resonance Raman responses from biomolecules and minerals. The 224.3 and 248.6 nm lasers elicit a fluorescence response from the aromatic amino and nucleic acids. Excitation at 325 nm may elicit activity from a variety of biomolecules, but is more likely to elicit mineral fluorescence. The resultant fluorescence images provide in situ chemical and morphological maps of microorganisms and the associated organic matrix. Visible broadband reflectance images provide orientation against the mineral background. Environmental scanning electron micrographs provided detailed morphological information. The technique has made possible the construction of detailed fluorescent maps extending from the surface of an Antarctic sandstone sample to a depth of 8.5 mm. The images detect no evidence of microbial life in the superficial 0.2 mm crustal layer. The black lichen component between 0.3 and 0.5 mm deep absorbs all wavelengths of both laser and broadband illumination. Filamentous deep ultraviolet native fluorescent activity dominates in the white layer between 0.6 mm and 5.0 mm from the surface. These filamentous forms are fungi that continue into the red (iron-rich) region of the sample extending from 5.0 to 8.5 mm. Using differential image subtraction techniques it is possible to identify fungal nuclei. The ultraviolet response is markedly attenuated in this region, apparently from the absorption of ultraviolet light by iron-rich particles coating

  19. Fluorescence quenching and photocatalytic degradation of textile dyeing waste water by silver nanoparticles

    Kavitha, S. R.; Umadevi, M.; Janani, S. R.; Balakrishnan, T.; Ramanibai, R.

    2014-06-01

    Silver nanoparticles (Ag NPs) of different sizes have been prepared by chemical reduction method and characterized using UV-vis spectroscopy and transmission electron microscopy (HRTEM). Fluorescence spectral analysis showed that the quenching of fluorescence of textile dyeing waste water (TDW) has been found to decrease with decrease in the size of the Ag NPs. Experimental results show that the silver nanoparticles can quench the fluorescence emission of adsorbed TDW effectively. The fluorescence interaction between Ag NPs (acceptor) and TDW (donor) confirms the Förster Resonance Energy Transfer (FRET) mechanism. Long range dipole-dipole interaction between the excited donor and ground state acceptor molecules is the dominant mechanism responsible for the energy transfer. Furthermore, photocatalytic degradation of TDW was measured spectrophotometrically by using silver as nanocatalyst under UV light illumination. The kinetic study revealed that synthesized Ag NPs was found to be effective in degrading TDW.

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

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

    2013-06-26

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

  1. In Vivo Deep Tissue Fluorescence and Magnetic Imaging Employing Hybrid Nanostructures.

    Ortgies, Dirk H; de la Cueva, Leonor; Del Rosal, Blanca; Sanz-Rodríguez, Francisco; Fernández, Nuria; Iglesias-de la Cruz, M Carmen; Salas, Gorka; Cabrera, David; Teran, Francisco J; Jaque, Daniel; Martín Rodríguez, Emma

    2016-01-20

    Breakthroughs in nanotechnology have made it possible to integrate different nanoparticles in one single hybrid nanostructure (HNS), constituting multifunctional nanosized sensors, carriers, and probes with great potential in the life sciences. In addition, such nanostructures could also offer therapeutic capabilities to achieve a wider variety of multifunctionalities. In this work, the encapsulation of both magnetic and infrared emitting nanoparticles into a polymeric matrix leads to a magnetic-fluorescent HNS with multimodal magnetic-fluorescent imaging abilities. The magnetic-fluorescent HNS are capable of simultaneous magnetic resonance imaging and deep tissue infrared fluorescence imaging, overcoming the tissue penetration limits of classical visible-light based optical imaging as reported here in living mice. Additionally, their applicability for magnetic heating in potential hyperthermia treatments is assessed.

  2. Distributed fluorescent optical fiber proximity sensor: Towards a proof of concept

    Gălătuș, Ramona; Faragó, Paul; Miluski, Piotr; Valles, Juan-Antonio

    2018-06-01

    Fluorescent fibers are optical fibers which emit light as a response to an incident phenomenon, usually an incident light. Operation depends on the doping dyes, which determine specific fluorescence and optical characteristics useful in the development of optical sensors. In this work we propose a low-cost distributed proximity sensor implemented using a red fluorescent fiber, to provide a security option for a surface plasmon resonance system. Operation of the proposed sensor relies on having the incident illumination intensity varied by the presence or absence of an obstacle in the vicinity of the sensing element. This will influence the radiated fluorescence accordingly. The proposed setup for the implementation of the optical proximity sensor assumes having a high brightness LED deployed for axial fiber illumination and a blue LED for side illumination. Electronic processing then accounts for gain and digitization. Measurement results of the prototype validate the proposed concept.

  3. Picocyanobacteria and deep-ocean fluorescent dissolved organic matter share similar optical properties

    Zhao, Zhao; Gonsior, Michael; Luek, Jenna; Timko, Stephen; Ianiri, Hope; Hertkorn, Norbert; Schmitt-Kopplin, Philippe; Fang, Xiaoting; Zeng, Qinglu; Jiao, Nianzhi; Chen, Feng

    2017-05-01

    Marine chromophoric dissolved organic matter (CDOM) and its related fluorescent components (FDOM), which are widely distributed but highly photobleached in the surface ocean, are critical in regulating light attenuation in the ocean. However, the origins of marine FDOM are still under investigation. Here we show that cultured picocyanobacteria, Synechococcus and Prochlorococcus, release FDOM that closely match the typical fluorescent signals found in oceanic environments. Picocyanobacterial FDOM also shows comparable apparent fluorescent quantum yields and undergoes similar photo-degradation behaviour when compared with deep-ocean FDOM, further strengthening the similarity between them. Ultrahigh-resolution mass spectrometry (MS) and nuclear magnetic resonance spectroscopy reveal abundant nitrogen-containing compounds in Synechococcus DOM, which may originate from degradation products of the fluorescent phycobilin pigments. Given the importance of picocyanobacteria in the global carbon cycle, our results indicate that picocyanobacteria are likely to be important sources of marine autochthonous FDOM, which may accumulate in the deep ocean.

  4. Comment on ’Single Pentacene Molecules Detected by Fluorescence Excitation in a P-Terphenyl Crystal’

    1990-12-10

    8217 NO 11 TITLE (include Security Classification) Comment on "Single Pentacene Molecules Detected by Fluorescence Excitation in a p-Terphenyl Crystal" 12...8217 {Continue on reverse it necessary and identify by block numboer) Using h--,Ihly efficient Fluorescence excitation spectroscov of individual pentacene ...molecular impurities in p-terphenvl crystals, we have observed that some pentacene defects exhibit spcntaneous spectral jumps in their resonance frequency at

  5. Cryo-imaging of fluorescently labeled single cells in a mouse

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

    2009-02-01

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

  6. Plasmonics Enhanced Smartphone Fluorescence Microscopy

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

    2017-01-01

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

  7. Fluorescence and phosphorescence of rutin

    Bondarev, Stanislav L., E-mail: bondarev@imaph.bas-net.by [Minsk State Higher Radioengineering College, 220005 Minsk (Belarus); Knyukshto, Valeri N. [B.I. Stepanov Institute of Physics, National Academy of Sciences of Belarus, 220072 Minsk (Belarus)

    2013-10-15

    Rutin is one of the most promising flavonoid from a pharmacological and biochemical point of view. Here we have explored its spectroscopic and photophysical properties at room temperature and 77 K using steady-state absorption-luminescence methods and pulse spectroscopy equipment. By excitation into the absorption band 1 of rutin in methanol at room temperature the normal Stokes' shifted fluorescence with a maximum at 415 nm and quantum yield of 2×10{sup −4} was revealed. However, by excitation into the bands 2 and 3 any emission wasn’t observed. At 77 K in ethanol glass we have observed fluorescence at 410 nm and phosphorescence at 540 nm for the first time. As a result the adequate energetic scheme including the lowest electronic excited singlet at 26000 cm{sup −1} and triplet at 19600 cm{sup −1} states was proposed. -- Highlights: • Rutin fluorescence and phosphorescence at 77 K were revealed for the first time. • Room temperature fluorescence is determined by maximum at 415 nm and yield of 2×10{sup −4}. • Violation of Vavilov–Kasha rule by excitation into the absorption bands 2 and 3. • Fluorescence and phosphorescence in rutin are caused by the allowed π, π{sup (⁎)} transitions.

  8. Plasmonics Enhanced Smartphone Fluorescence Microscopy

    Wei, Qingshan

    2017-05-12

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

  9. Controlling Parametric Resonance

    Galeazzi, Roberto; Pettersen, Kristin Ytterstad

    2012-01-01

    the authors review the conditions for the onset of parametric resonance, and propose a nonlinear control strategy in order to both induce the resonant oscillations and to stabilize the unstable motion. Lagrange’s theory is used to derive the dynamics of the system and input–output feedback linearization...

  10. Electron Paramagnetic Resonance Spectroscopy

    Home; Journals; Resonance – Journal of Science Education; Volume 20; Issue 11. Electron Paramagnetic Resonance Spectroscopy: Biological Applications. B G Hegde. General Article Volume 20 Issue 11 November 2015 pp 1017-1032. Fulltext. Click here to view fulltext PDF. Permanent link:

  11. Electromagnetic resonance waves

    Villaba, J.M.; Manjon, F.J.; Guirao, A.; Andres, M.V.

    1994-01-01

    We describe in this paper a set of experiments designed to make qualitative and quantitative measurements on electromagnetic resonances of several simple systems. The experiments are designed for the undergraduate laboratory of Electricity and Magnetism in Physics. These experiments can help the students understanding the concept of resonance, which appears in different fields of Physics. (Author) 8 refs

  12. Laser magnetic resonance spectroscopy

    Ferrari, C.A.

    1985-01-01

    The technique of laser resonance magnetic resonance allows one to study the high-resolution spectroscopy of transient paramagnetic species, viz, atoms, radicals, and molecular ions. This article is a brief exposition of the method, describing the principles, instrumentation and applicability of the IR and FIR-LMR and shows results of HF + . (Author) [pt

  13. Resonance and Fractal Geometry

    Broer, Henk W.

    The phenomenon of resonance will be dealt with from the viewpoint of dynamical systems depending on parameters and their bifurcations. Resonance phenomena are associated to open subsets in the parameter space, while their complement corresponds to quasi-periodicity and chaos. The latter phenomena

  14. Nuclear Magnetic Resonance Spectroscopy

    Home; Journals; Resonance – Journal of Science Education; Volume 9; Issue 1. Nuclear Magnetic Resonance Spectroscopy. Susanta Das. General Article Volume 9 Issue 1 January 2004 pp 34-49. Fulltext. Click here to view fulltext PDF. Permanent link: https://www.ias.ac.in/article/fulltext/reso/009/01/0034-0049. Keywords.

  15. Cardiac magnetic resonance imaging

    2011-03-06

    Mar 6, 2011 ... Cardiac magnetic resonance imaging. Cardiovascular magnetic resonance imaging is becoming a routine diagnostic technique. BRUCE s sPOTTiswOOdE, PhD. MRC/UCT Medical Imaging Research Unit, University of Cape Town, and Division of Radiology, Stellenbosch University. Bruce Spottiswoode ...

  16. Gold and silver nanoparticles based superquenching of fluorescence: A review

    Ghosh, Debanjana; Chattopadhyay, Nitin, E-mail: nitin.chattopadhyay@yahoo.com

    2015-04-15

    The short review highlights the recent advances on the gold and silver nanoparticles induced efficient quenching of fluorescence from various fluorophores looking at their promising use as optical rulers and chemo-/bio- sensors. The fluorescence quenching often leads to the increase in the Stern–Volmer constant (K{sub SV}~10{sup 7}–10{sup 10} mol{sup −1} dm{sup 3}) several orders of magnitude higher than the values observed for the normal photochemical quenching processes (~10{sup 2} mol{sup −1} dm{sup 3}). This amplified quenching has been termed as “super-quenching” or “hyper-quenching”. Energy transfer (ET) is established from the donor to the metal nanoparticles rationalizing these fast quenching processes. Considering the distance dependence of the ET process, Förster resonance energy transfer (FRET) and nanometal surface energy transfer (NSET) are ascribed to take place. These sensitive distance dependent phenomena serve as the spectroscopic ruler to measure the intra- or intermolecular distances between the interacting partners. In this account focus has been laid on the size dependent energy transfer and super- and hyper- quenching of the fluorescence of the donor moieties by the nanometals and their probable applications in sensing. Rationalization has been made for the nanoparticle induced huge enhancement in the quenching efficiency. The impact of this review lies in the possible application of these amplified quenching processes in designing high sensitive chemical and biological sensors. - Highlights: • Super efficient quenching of fluorescence of probes by gold and silver nanoparticles is highlighted. • The amplified fluorescence quenching of dyes and polymers is rationalized. • Energy transfer is assigned to be responsible for the efficient quenching process. • Amplified quenching has its potential use in designing sensitive chemical/biological sensors.

  17. Gold and silver nanoparticles based superquenching of fluorescence: A review

    Ghosh, Debanjana; Chattopadhyay, Nitin

    2015-01-01

    The short review highlights the recent advances on the gold and silver nanoparticles induced efficient quenching of fluorescence from various fluorophores looking at their promising use as optical rulers and chemo-/bio- sensors. The fluorescence quenching often leads to the increase in the Stern–Volmer constant (K SV ~10 7 –10 10 mol −1 dm 3 ) several orders of magnitude higher than the values observed for the normal photochemical quenching processes (~10 2 mol −1 dm 3 ). This amplified quenching has been termed as “super-quenching” or “hyper-quenching”. Energy transfer (ET) is established from the donor to the metal nanoparticles rationalizing these fast quenching processes. Considering the distance dependence of the ET process, Förster resonance energy transfer (FRET) and nanometal surface energy transfer (NSET) are ascribed to take place. These sensitive distance dependent phenomena serve as the spectroscopic ruler to measure the intra- or intermolecular distances between the interacting partners. In this account focus has been laid on the size dependent energy transfer and super- and hyper- quenching of the fluorescence of the donor moieties by the nanometals and their probable applications in sensing. Rationalization has been made for the nanoparticle induced huge enhancement in the quenching efficiency. The impact of this review lies in the possible application of these amplified quenching processes in designing high sensitive chemical and biological sensors. - Highlights: • Super efficient quenching of fluorescence of probes by gold and silver nanoparticles is highlighted. • The amplified fluorescence quenching of dyes and polymers is rationalized. • Energy transfer is assigned to be responsible for the efficient quenching process. • Amplified quenching has its potential use in designing sensitive chemical/biological sensors

  18. Fundamentals of nanomechanical resonators

    Schmid, Silvan; Roukes, Michael Lee

    2016-01-01

    This authoritative book introduces and summarizes the latest models and skills required to design and optimize nanomechanical resonators, taking a top-down approach that uses macroscopic formulas to model the devices. The authors cover the electrical and mechanical aspects of nano electromechanical system (NEMS) devices. The introduced mechanical models are also key to the understanding and optimization of nanomechanical resonators used e.g. in optomechanics. Five comprehensive chapters address: The eigenmodes derived for the most common continuum mechanical structures used as nanomechanical resonators; The main sources of energy loss in nanomechanical resonators; The responsiveness of micro and nanomechanical resonators to mass, forces, and temperature; The most common underlying physical transduction mechanisms; The measurement basics, including amplitude and frequency noise. The applied approach found in this book is appropriate for engineering students and researchers working with micro and nanomechanical...

  19. Resonant snubber inverter

    Lai, Jih-Sheng; Young, Sr., Robert W.; Chen, Daoshen; Scudiere, Matthew B.; Ott, Jr., George W.; White, Clifford P.; McKeever, John W.

    1997-01-01

    A resonant, snubber-based, soft switching, inverter circuit achieves lossless switching during dc-to-ac power conversion and power conditioning with minimum component count and size. Current is supplied to the resonant snubber branches solely by the main inverter switches. Component count and size are reduced by use of a single semiconductor switch in the resonant snubber branches. Component count is also reduced by maximizing the use of stray capacitances of the main switches as parallel resonant capacitors. Resonance charging and discharging of the parallel capacitances allows lossless, zero voltage switching. In one embodiment, circuit component size and count are minimized while achieving lossless, zero voltage switching within a three-phase inverter.

  20. The Tracking Resonance Frequency Method for Photoacoustic Measurements Based on the Phase Response

    Suchenek, Mariusz

    2017-04-01

    One of the major issues in the use of the resonant photoacoustic cell is the resonance frequency of the cell. The frequency is not stable, and its changes depend mostly on temperature and gas mixture. This paper presents a new method for tracking resonance frequency, where both the amplitude and phase are calculated from the input samples. The stimulating frequency can be adjusted to the resonance frequency of the cell based on the phase. This method was implemented using a digital measurement system with an analog to digital converter, field programmable gate array (FPGA) and a microcontroller. The resonance frequency was changed by the injection of carbon dioxide into the cell. A theoretical description and experimental results are also presented.

  1. Label free selective detection of estriol using graphene oxide-based fluorescence sensor

    Kushwaha, H. S.; Sao, Reshma; Vaish, Rahul

    2014-07-01

    Water-soluble and fluorescent Graphene oxide (GO) is biocompatible, easy, and economical to synthesize. Interestingly, GO is also capable of quenching fluorescence. On the basis of its fluorescence and quenching abilities, GO has been reported to serve as an energy acceptor in a fluorescence resonance energy transfer (FRET) sensor. GO-based FRET biosensors have been widely reported for sensing of proteins, nucleic acid, ATP (Adenosine triphosphate), etc. GO complexes with fluorescent dyes and enzymes have been used to sense metal ions. Graphene derivatives have been used for sensing endocrine-disrupting chemicals like bisphenols and chlorophenols with high sensitivity and good reproducibility. On this basis, a novel GO based fluorescent sensor has been successfully designed to detect estriol with remarkable selectivity and sensitivity. Estriol is one of the three estrogens in women and is considered to be medically important. Estriol content of maternal urine or plasma acts as an important screening marker for estimating foetal growth and development. In addition, estriol is also used as diagnostic marker for diseases like breast cancer, osteoporosis, neurodegenerative and cardiovascular diseases, insulin resistance, lupus erythematosus, endometriosis, etc. In this present study, we report for the first time a rapid, sensitive with detection limit of 1.3 nM, selective and highly biocompatible method for label free detection of estriol under physiological conditions using fluorescence assay.

  2. DNA-hosted copper nanoclusters/graphene oxide based fluorescent biosensor for protein kinase activity detection.

    Wang, Mengke; Lin, Zihan; Liu, Qing; Jiang, Shan; Liu, Hua; Su, Xingguang

    2018-07-05

    A novel fluorescent biosensor for protein kinase activity (PKA) detection was designed by applying double-strands DNA-hosted copper nanoclusters (dsDNA-CuNCs) and graphene oxide (GO). One DNA strand of the dsDNA consisted of two domains, one domain can hybridize with another complementary DNA strand to stabilize the fluorescent CuNCs and another domain was adenosine 5'-triphosphate (ATP) aptamer. ATP aptamer of the dsDNA-CuNCs would be spontaneously absorbed onto the GO surface through π-π stacking interactions. Thus GO can efficiently quench the fluorescence (FL) of dsDNA-CuNCs through fluorescence resonance energy transfer (FRET). In the present of ATP, ATP specifically combined with ATP aptamer to form ATP-ATP aptamer binding complexes, which had much less affinity to GO, resulting in the fluorescence recovery of the system. Nevertheless, in the presence of PKA, ATP could be translated into ADP and ADP could not combine with ATP aptamer resulting in the fluorescence quenching of dsDNA-CuNCs again. According to the change of the fluorescence signal, PKA activity could be successfully monitored in the range of 0.1-5.0 U mL -1 with a detection limit (LOD) of 0.039 U mL -1 . Besides, the inhibitory effect of H-89 on PKA activity was studied. The sensor was performed for PKA activity detection in cell lysates with satisfactory results. Copyright © 2018 Elsevier B.V. All rights reserved.

  3. Dual-Color Fluorescence Imaging of Magnetic Nanoparticles in Live Cancer Cells Using Conjugated Polymer Probes

    Sun, Minjie; Sun, Bin; Liu, Yun; Shen, Qun-Dong; Jiang, Shaojun

    2016-01-01

    Rapid growth in biological applications of nanomaterials brings about pressing needs for exploring nanomaterial-cell interactions. Cationic blue-emissive and anionic green-emissive conjugated polymers are applied as dual-color fluorescence probes to the surface of negatively charged magnetic nanoparticles through sequentially electrostatic adsorption. These conjugated polymers have large extinction coefficients and high fluorescence quantum yield (82% for PFN and 62% for ThPFS). Thereby, one can visualize trace amount (2.7 μg/mL) of fluorescence-labeled nanoparticles within cancer cells by confocal laser scanning microscopy. Fluorescence labeling by the conjugated polymers is also validated for quantitative determination of the internalized nanoparticles in each individual cell by flow cytometry analysis. Extensive overlap of blue and green fluorescence signals in the cytoplasm indicates that both conjugated polymer probes tightly bind to the surface of the nanoparticles during cellular internalization. The highly charged and fluorescence-labeled nanoparticles non-specifically bind to the cell membranes, followed by cellular uptake through endocytosis. The nanoparticles form aggregates inside endosomes, which yields a punctuated staining pattern. Cellular internalization of the nanoparticles is dependent on the dosage and time. Uptake efficiency can be enhanced three-fold by application of an external magnetic field. The nanoparticles are low cytotoxicity and suitable for simultaneously noninvasive fluorescence and magnetic resonance imaging application. PMID:26931282

  4. Demonstration of a visual cell-based assay for screening glucose ...

    GLUT4 translocation is visualized by live cell imaging based on GFP fluorescence by employing a cooled charge-coupled device camera attached to a fluorescent microscope. This video imaging method and further quantitative analysis of GLUT4 on the cell membrane provide rapid and foolproof visual evidence that this ...

  5. Fluorescence confocal microscopy for pathologists.

    Ragazzi, Moira; Piana, Simonetta; Longo, Caterina; Castagnetti, Fabio; Foroni, Monica; Ferrari, Guglielmo; Gardini, Giorgio; Pellacani, Giovanni

    2014-03-01

    Confocal microscopy is a non-invasive method of optical imaging that may provide microscopic images of untreated tissue that correspond almost perfectly to hematoxylin- and eosin-stained slides. Nowadays, following two confocal imaging systems are available: (1) reflectance confocal microscopy, based on the natural differences in refractive indices of subcellular structures within the tissues; (2) fluorescence confocal microscopy, based on the use of fluorochromes, such as acridine orange, to increase the contrast epithelium-stroma. In clinical practice to date, confocal microscopy has been used with the goal of obviating the need for excision biopsies, thereby reducing the need for pathological examination. The aim of our study was to test fluorescence confocal microscopy on different types of surgical specimens, specifically breast, lymph node, thyroid, and colon. The confocal images were correlated to the corresponding histological sections in order to provide a morphologic parallel and to highlight current limitations and possible applications of this technology for surgical pathology practice. As a result, neoplastic tissues were easily distinguishable from normal structures and reactive processes such as fibrosis; the use of fluorescence enhanced contrast and image quality in confocal microscopy without compromising final histologic evaluation. Finally, the fluorescence confocal microscopy images of the adipose tissue were as accurate as those of conventional histology and were devoid of the frozen-section-related artefacts that can compromise intraoperative evaluation. Despite some limitations mainly related to black/white images, which require training in imaging interpretation, this study confirms that fluorescence confocal microscopy may represent an alternative to frozen sections in the assessment of margin status in selected settings or when the conservation of the specimen is crucial. This is the first study to employ fluorescent confocal microscopy on

  6. Nuclear Magnetic Resonance spectroscopy studies of proteins-glycoconjugates interactions

    Marchetti, Roberta

    2013-01-01

    This PhD thesis work has been focused on the analysis of the structural requisites for recognition and binding between proteins and glycoconjugates, essential for the comprehension of mechanisms of paramount importance in chemistry, biology and biomedicine. A large variety of techniques, such as crystallographic analysis, titration microcalorimetry (ITC), surface plasmon resonance (SPR) and fluorescence spectroscopy, allows the elucidation of molecular recognition events. In the last years...

  7. Using a Spectrofluorometer for Resonance Raman Spectra of Organic Molecules

    Vadivel Masilamani

    2017-01-01

    Full Text Available Scattering (Rayleigh and Raman and fluorescence are two common light signals that frequently occur together, confusing the researchers and graduate students experimenting in molecular spectroscopy laboratories. This report is a brief study presenting a clear discrimination between the two signals mentioned, employing a common spectrofluorometer such as the PerkinElmer LS 55. Even better, the resonance Raman signal of a molecule (e.g., acetone can be obtained elegantly using the same instrument.

  8. Fluorescence detection of dental calculus

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

    2010-11-01

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

  9. Fluorescence Quenching of Alpha-Fetoprotein by Gold Nanoparticles: Effect of Dielectric Shell on Non-Radiative Decay

    Zhu, Jian; Li, Jian-Jun; Wang, A.-Qing; Chen, Yu; Zhao, Jun-Wu

    2010-09-01

    Fluorescence quenching spectrometry was applied to study the interactions between gold colloidal nanoparticles and alpha-fetoprotein (AFP). Experimental results show that the gold nanoparticles can quench the fluorescence emission of adsorbed AFP effectively. Furthermore, the intensity of fluorescence emission peak decreases monotonously with the increasing gold nanoparticles content. A mechanism based on surface plasmon resonance-induced non-radiative decay was investigated to illuminate the effect of a dielectric shell on the fluorescence quenching ability of gold nanoparticles. The calculation results show that the increasing dielectric shell thickness may improve the monochromaticity of fluorescence quenching. However, high energy transfer efficiency can be obtained within a wide wavelength band by coating a thinner dielectric shell.

  10. Advances in magnetic resonance 10

    Waugh, John S

    2013-01-01

    Advances in Magnetic Resonance, Volume 10, presents a variety of contributions to the theory and practice of magnetic resonance. The book contains three chapters that examine superoperators in magnetic resonance; ultrasonically modulated paramagnetic resonance; and the utility of electron paramagnetic resonance (EPR) and electron-nuclear double-resonance (ENDOR) techniques for studying low-frequency modes of atomic fluctuations and their significance for understanding the mechanism of structural phase transitions in solids.

  11. High content screening for G protein-coupled receptors using cell-based protein translocation assays

    Grånäs, Charlotta; Lundholt, Betina Kerstin; Heydorn, Arne

    2005-01-01

    G protein-coupled receptors (GPCRs) have been one of the most productive classes of drug targets for several decades, and new technologies for GPCR-based discovery promise to keep this field active for years to come. While molecular screens for GPCR receptor agonist- and antagonist-based drugs...... will continue to be valuable discovery tools, the most exciting developments in the field involve cell-based assays for GPCR function. Some cell-based discovery strategies, such as the use of beta-arrestin as a surrogate marker for GPCR function, have already been reduced to practice, and have been used...... as valuable discovery tools for several years. The application of high content cell-based screening to GPCR discovery has opened up additional possibilities, such as direct tracking of GPCRs, G proteins and other signaling pathway components using intracellular translocation assays. These assays provide...

  12. Multiple photon resonances

    Elliott, C.J.; Feldman, B.J.

    1979-02-01

    A detailed theoretical analysis is presented of the interaction of intense near-resonant monochromatic radiation with an N-level anharmonic oscillator. In particular, the phenomenon of multiple photon resonance, the process by which an N-level system resonantly absorbs two or more photons simultaneously, is investigated. Starting from the Schroedinger equation, diagrammatic techniques are developed that allow the resonant process to be analyzed quantitatively, in analogy with well-known two-level coherent phenomena. In addition, multiple photon Stark shifts of the resonances, shifts absent in two-level theory, are obtained from the diagrams. Insights into the nature of multiple photon resonances are gained by comparing the quantum mechanical system with classical coupled pendulums whose equations of motion possess identical eigenvalues and eigenvectors. In certain limiting cases, including that of the resonantly excited N-level harmonic oscillator and that of the equally spaced N-level system with equal matrix elements, analytic results are derived. The influence of population relaxation and phase-disrupting collisions on the multiple photon process are also analyzed, the latter by extension of the diagrammatic technique to the density matrix equations of motion. 11 figures

  13. Properties of spiral resonators

    Haeuser, J.

    1989-10-01

    The present thesis deals with the calculation and the study of the application possibilities of single and double spiral resonators. The main aim was the development and the construction of reliable and effective high-power spiral resonators for the UNILAC of the GSI in Darmstadt and the H - -injector for the storage ring HERA of DESY in Hamburg. After the presentation of the construction and the properties of spiral resonators and their description by oscillating-circuit models the theoretical foundations of the bunching are presented and some examples of a rebuncher and debuncher and their influence on the longitudinal particle dynamics are shown. After the description of the characteristic accelerator quantities by means of an oscillating-circuit model and the theory of an inhomogeneous λ/4 line it is shown, how the resonance frequency and the efficiency of single and double spiral resonators can be calculated from the geometrical quantities of the structure. In the following the dependence of the maximal reachable resonator voltage in dependence on the gap width and the surface of the drift tubes is studied. Furthermore the high-power resonators are presented, which were built for the different applications for the GSI in Darmstadt, DESY in Hamburg, and for the FOM Institute in Amsterdam. (orig./HSI) [de

  14. In vivo tomographic imaging with fluorescence and MRI using tumor-targeted dual-labeled nanoparticles

    Zhang Y

    2013-12-01

    Full Text Available Yue Zhang,1 Bin Zhang,1 Fei Liu,1,2 Jianwen Luo,1,3 Jing Bai1 1Department of Biomedical Engineering, School of Medicine, 2Tsinghua-Peking Center for Life Sciences, 3Center for Biomedical Imaging Research, Tsinghua University, Beijing, People's Republic of China Abstract: Dual-modality imaging combines the complementary advantages of different modalities, and offers the prospect of improved preclinical research. The combination of fluorescence imaging and magnetic resonance imaging (MRI provides cross-validated information and direct comparison between these modalities. Here, we report on the application of a novel tumor-targeted, dual-labeled nanoparticle (NP, utilizing iron oxide as the MRI contrast agent and near infrared (NIR dye Cy5.5 as the fluorescent agent. Results of in vitro experiments verified the specificity of the NP to tumor cells. In vivo tumor targeting and uptake of the NPs in a mouse model were visualized by fluorescence and MR imaging collected at different time points. Quantitative analysis was carried out to evaluate the efficacy of MRI contrast enhancement. Furthermore, tomographic images were also acquired using both imaging modalities and cross-validated information of tumor location and size between these two modalities was revealed. The results demonstrate that the use of dual-labeled NPs can facilitate the dual-modal detection of tumors, information cross-validation, and direct comparison by combing fluorescence molecular tomography (FMT and MRI. Keywords: dual-modality, fluorescence molecular tomography (FMT, magnetic resonance imaging (MRI, nanoparticle

  15. Magnetic Resonance Force Microscopy System

    Federal Laboratory Consortium — The Magnetic Resonance Force Microscopy (MRFM) system, developed by ARL, is the world's most sensitive nuclear magnetic resonance (NMR) spectroscopic analysis tool,...

  16. Fluorescence Molecular Tomography: Principles and Potential for Pharmaceutical Research

    Florian Stuker

    2011-04-01

    Full Text Available Fluorescence microscopic imaging is widely used in biomedical research to study molecular and cellular processes in cell culture or tissue samples. This is motivated by the high inherent sensitivity of fluorescence techniques, the spatial resolution that compares favorably with cellular dimensions, the stability of the fluorescent labels used and the sophisticated labeling strategies that have been developed for selectively labeling target molecules. More recently, two and three-dimensional optical imaging methods have also been applied to monitor biological processes in intact biological organisms such as animals or even humans. These whole body optical imaging approaches have to cope with the fact that biological tissue is a highly scattering and absorbing medium. As a consequence, light propagation in tissue is well described by a diffusion approximation and accurate reconstruction of spatial information is demanding. While in vivo optical imaging is a highly sensitive method, the signal is strongly surface weighted, i.e., the signal detected from the same light source will become weaker the deeper it is embedded in tissue, and strongly depends on the optical properties of the surrounding tissue. Derivation of quantitative information, therefore, requires tomographic techniques such as fluorescence molecular tomography (FMT, which maps the three-dimensional distribution of a fluorescent probe or protein concentration. The combination of FMT with a structural imaging method such as X-ray computed tomography (CT or Magnetic Resonance Imaging (MRI will allow mapping molecular information on a high definition anatomical reference and enable the use of prior information on tissue’s optical properties to enhance both resolution and sensitivity. Today many of the fluorescent assays originally developed for studies in cellular systems have been successfully translated for experimental studies in animals. The opportunity of monitoring molecular

  17. Resonant power converters

    Kazimierczuk, Marian K

    2012-01-01

    This book is devoted to resonant energy conversion in power electronics. It is a practical, systematic guide to the analysis and design of various dc-dc resonant inverters, high-frequency rectifiers, and dc-dc resonant converters that are building blocks of many of today's high-frequency energy processors. Designed to function as both a superior senior-to-graduate level textbook for electrical engineering courses and a valuable professional reference for practicing engineers, it provides students and engineers with a solid grasp of existing high-frequency technology, while acquainting them wit

  18. Excitation of Nucleon Resonances

    Burkert, Volker D.

    2001-01-01

    I discuss developments in the area of nucleon resonance excitation, both necessary and feasible, that would put our understanding of nucleon structure in the regime of strong QCD on a qualitatively new level. They involve the collection of high quality data in various channels, a more rigorous approach in the search for ''missing'' resonances, an effort to compute some critical quantities in nucleon resonance excitations from first principles, i.e. QCD, and a proposal focused to obtain an understanding of a fundamental quantity in nucleon structure

  19. Dihadronic and dileptonic resonances

    Gareev, F.A.; Barabanov, M.Yu.; Kazacha, G.S.

    1997-01-01

    Simple phenomenological rules are suggested for calculation of dihadron and dilepton resonance masses. A general interpretation is given for different exotic resonances in nuclear physics: Darmstadt-effect, dibaryon, dipion and other resonances. Information about the inner structure of e ± , proton, neutron, pions and so on can be obtained from the usual reactions of the type e + + e - =>γγ, e ± +γ=>e ± γ, e ± μ ± , e ± N... at low, intermediate and high energies using existing experimental devices

  20. Multiquark resonant states

    Shahbazian, B.A.

    1982-01-01

    The invariant mass spectra of forty nine hadronic systems with hypercharge, strangeness and baryon number, varied in wide limits have been studied. Resonance peaks have been found in the invariant mass spectra of Y 2 and #betta#pπ 2495 MeV/c 2 resonant states. Three more candidates for anti qq 4 states were found #bettaπ# + π + : 1705, 2072, 2605 MeV/c 2 . The masses of all these candidates are in good agreement with Bag Model predictions. A hypercharge selection rule is suggested: ''The hypercharge of hadronic resonances in weak gravitational fields cannot exceed one Y <= 1