Homogeneous time-resolved fluoroimmunoassay of bensulfuron-methyl by using terbium fluorescence energy transfer

A sensitive homogenous time-resolved fluoroimmunoassay (TR-FIA) method for bensulfuron-methyl (BSM) based on fluorescence resonance energy transfer (FRET) from a Tb3+ fluorescent chelate with N,N,N',N'-[2,6-bis(3'-aminomethyl-1'-pyrazoly)-4-phenylpyridine] tetrakis(acetic acid) (BPTA-Tb3+) to organic dye, Cy3 or Cy3.5 has been developed. New method combined the use of BPTA-Tb3+ labeled streptavidin, Cy3 or Cy3.5 labeled anti-BSM monoclonal antibody and biotinylated BSM-BSA conjugate (BSA is bovine serum albumin) for competitive-type immunoassay. After BPTA-Tb3+ labeled streptavidin was reacted with a competitive immune reaction solution containing biotinylated BSM-BSA, BSM sample and Cy3 or Cy3.5 labeled anti-BSM monoclonal antibody, the sensitized and long-lived emission of Cy3 or Cy3.5 derived from FRET was measured, and thus the concentration of BSM in sample was calculated. The present method has the advantages of rapidity, simplicity and high sensitivity since the B/F (bound reagent/free reagent) separation steps and the solid-phase carrier are not necessary. The method gives the detection limit of 2.10 ng ml(-1). The coefficient variations of the method are less than 1.5% and the recoveries are in the range of 95-105% for BSM water sample measurement. (C) 2001 Elsevier Science B.V. All rights reserved.

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

Macrophage differentiation antigen associated with complement three receptor function (Mac-1) belongs to beta(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. (c) 2006 Elsevier Inc. All rights reserved.

Detection of constitutive homomeric associations of the integrins Mac-1 subunits by fluorescence resonance energy transfer in living cells

Integrins alpha(M)beta(2) plays important role on leukocytes, such as adhesion, migration, phagocytosis, and apoptosis. It was hypothesized that homomeric associations of integrin subunits provide a driving force for integrins activation, and simultaneously inducing the formation of integrins clusters. However, experimental reports on homomeric associations between integrin subunits are still controversial. Here, we proved the homomeric associations of the isolated Mac-1 subunits in living cells using three-channel fluorescence resonance energy transfer (FRET) microscopy and FRET spectra methods. We found that the extent of homomeric associations between beta(2) subunits is higher than alpha(M) subunits. Furthermore, FRET imaging indicated that the extent of homomeric associations of the Mac-1 subunits is higher along the plasma membrane than in the cytoplasm. Finally, we suggested that homomeric associations of the transmernbrane domains or/and cytoplasmic domains may provide the driving force for the formation of constitutive homomeric associations between alpha(M) or beta(2) subunits. (c) 2006 Elsevier Inc. All rights reserved.

Energy transfer and infrared-to-visible upconversion luminescence of Er3+/Yb3+-codoped halide modified tellurite glasses

We report on the energy transfer and frequency upconversion spectroscopic properties of Er3+-doped and Er3+/Yb3+-codoped TeO2-ZnO-Na2O-PbCl2 halide modified tellurite glasses upon excitation with 808 and 978 nm laser diode. Three intense emissions centered at around 529, 546 and 657 nm, alongwith a very weak blue emission at 4 10 nm have clearly been observed for the Er3+/Yb3+-codoped halide modified tellurite glasses upon excitation at 978 nm and the involved mechanisms are explained. The quadratic dependence of fluorescence on excitation laser power confirms the fact that the two-photon contribute to the infrared to green-red upconversion emissions. And the blue upconversion at 410 nm involved a sequential three-photon absorption process. (c) 2005 Elsevier Ltd. All rights reserved.

Enhanced photoluminescence of Nd2O3 nanoparticles modified with silane-coupling agent: Fluorescent resonance energy transfer analysis

A liquid laser medium with a lifetime of 492 mu s and a fluorescent quantum efficiency of 52.5% has been presented by stably dispersing dimethyl dichorosilane-modified Nd2O3 nanoparticles in dimethylsulfoxide. Its optical properties and mechanism were investigated and explained by fluorescence resonance energy transfer theory. The calculation result shows that the quenching of Nd-III F-4(3/2)-> I-4(11/2) transition via O-H vibrational excitation can be eventually neglected. The main reason is that the silane-coupling agent molecules remove the -OH groups on Nd2O3 nanoparticles and form a protective out layer. (c) 2007 American Institute of Physics.

Luminescence and energy transfer of organically modified silica xerogels (OMSX) doped and undoped with Eu3+ and Tb3+

Organically modified silica xerogels (OMSX) and Eu3+ (Tb3+)-doped OMSX were prepared by the reaction of (3-aminopropyl) triethoxysilane (APS) with 3-isocyanatepropyltriethoxysilane (ICPTES) followed by the subsequent hydrolysis and condensation in the presence of Eu3+ (Tb3+) via sol-gel method, which were characterized by FT-IR, XRD, fluorescence excitation and emission spectra. The as-formed OMSX shows a strong blue emission with the maximum excitation and emission wavelength at 351 and 420 nm, respectively. Due to the spectral overlap between the emission band of OMSX and f-f absorption lines of Eu3+ and Tb3+ in the UV-blue region, an energy transfer was observed from OMSX host to Eu3+ and Tb3+ in OMSX/Eu3+ and OMSX/Tb3+, respectively. Excitation at 350-360 nm resulted in a very weak emission around 420 nm from OMSX host and strong emission of Eu3+ and Tb3+ in OMSX/Eu3+ and OMSX/Tb3+, respectively. The emission spectra of Eu3+ and Tb3+ consist of D-5(0)-F-7(J) (J = 0, 1, 2, 3, 4) and D-5(4)-F-7(J) (J = 6, 5, 4, 3), respectively. Furthermore, the predicted structure of OMSX/Eu3+ and OMSX/Tb3+ is presented.

Effective energy transfer and luminescence of LB films based on europium-substituted heteropolytungstate

The europium-substituted heteropolytungstate K13Eu(SiW11O39)(2) was successfully assembled into two lipids by LB technique for the first time. X-ray diffraction has shown a well defined lamellar for the LB films. The LB films have been characterized by fluorescence spectra and the characteristic luminescence behaviors were discussed. The ligand-metal charge transfer band could be observed in the spectra of the LB films, which could not be found in that of heteropolytungstate solid. The results of fluorescence spectra indicate the energy could be effectively transferred from ligands to the Eu3+ ions in the LB films and the luminescence efficiency was increased greatly. The influences of various lipids on the luminescence of polyoxometalates were investigated. The various interactions between monolayer and polyanions have different effect on the luminescence properties of europium-substituted heteropolytungstate.

Synthesis, luminescence, energy transfer and Langmuir-Blodgett films of amphiphilic complexes of rare earths with monooctadecyl phthalate

Three new amphiphilic rare earth complexes with only two organic long chains Ln (MOP)(2)Cl (MOP=monooctadecyl phthalate, Ln=Eu, Tb, Gd) were synthesized and characterized by elemental analysis. The complexes (Eu, Tb) showed good luminescence property with long fluorescence lifetime, whereas the intensity and lifetime of Tb complex are greater than those of Eu complex, By measuring the triplet energy levels of ligand based on energy transfer mechanism, above phenomena have been well explained. The Langmuir films of the complexes on the air/water interface were also studied and the results show that all of them have good film-forming property.

Nonradiative energy transfer study on the interface of compatibilized linear low density polyethylene/poly(methyl methacrylate) blends

Blends of chromophore-labeled LLDPE and chromophore-labeled PMMA compatibilized by block copolymer of hydrogenated polybutadiene and methyl methacrylate (PHB-b-PMMA) were studied by nonradiative energy transfer (NRET) technique. The ratio of fluorescence intensity of the donor at 336 nm and the acceptor at 408 nm (I-D/I-A) decreased with an increase in block copolymer content. At about 8 wt.-% block copolymer content I-D/I-A reached a minimum value, indicating the interdiffusion of LLDPE chains and PMMA chains in the interface is strongest. The influence of temperature on the interdiffusion of polymer chains in the interface was also examined. Samples quenched in liquid nitrogen from 140 degrees C showed lower energy transfer efficiencies than those annealed from 150 degrees C to room temperature.

INVESTIGATIONS ON THE ENERGY-TRANSFER OF EXCITED PARTICLE COLLISION WITH DOUBLE LASER LINES FOR EXCITATION OF LA(I)

Generally speaking, productions of thermally-assisted and stepwise fluorescence are the consequence of energy transfer caused by particle collision. In some circumstances, energy transfer caused by particle collision is considerably intense. We have ever used the fluorescence produced by energy transfer of particle collision to measure the branching ratios in the atomic transitions and acquired good results. To our knowledge, the systematic in

Energy transfer between silicon-oxygen-related defects and Yb3+ in transparent glass ceramics containing Ba2TiSi2O 8 nanocrystals

We report on the conversion of near-ultraviolet radiation of 250-350 nm into near-infrared emission of 970-1100 nm in Yb3+-doped transparent glass ceramics containing Ba2TiSi2O8 nanocrystals due to the energy transfer from the silicon-oxygen-related defects to Yb3+ ions. Efficient Yb3+ emission (F-2(5/2)-> F-2(7/2)) was detected under the excitation of defects absorption at 314 nm. The occurrence of energy transfer is proven by both steady state and time-resolved emission spectra, respectively, at 15 K. The Yb2O3 concentration dependent energy transfer efficiency has also been evaluated, and the maximum value is 65% for 8 mol % Yb2O3 doped glass ceramic. These materials are promising for the enhancement of photovoltaic conversion efficiency of silicon solar cells via spectra modification.