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Colourless crystalline solid coumarin with a bitter taste and sweet vanilla-like odor mostly acts as chemical protection against predators in plants. Anticoagulants (blood thinners), anti-fungicidal, anti-tumor and anti-inflammatory properties of coumarin are all used to treat skin diseases. Because of internal charge transfer (ICT), coumarin molecules displayed a solvatochromic effect in various solvents of varying polarity. The emission wavelength dependent fluorescence intensity was also affected by pH., (© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.)

The intrinsic fluorescence of nucleic acids is extremely weak compared to that of the fluorescent labels used to probe their structural and functional behavior. Thus, for technical reasons, the investigation of the intrinsic DNA fluorescence was limited for a long time. But with the improvement in spectroscopic techniques, the situation started to change around the turn of the century. During the past two decades, various factors modulating the static and dynamic properties of the DNA fluorescence have been determined; it was shown that, under certain conditions, quantum yields may be up 100 times higher than what was known so far. The ensemble of these studies opened up new paths for the development of label-free DNA fluorescence for biochemical applications. In parallel, these studies have shed new light on the primary processes leading to photoreactions that damage DNA when it absorbs UV radiation.We have been studying a variety of DNA systems, ranging from the monomeric nucleobases to double-stranded and four-stranded structures using fluorescence spectroscopy. The specificity of our work resides in the quantitative association of the steady-state fluorescence spectra with time-resolved data recorded from the femtosecond to the nanosecond timescales, made possible by the development of specific methodologies.Among others, our fluorescence studies provide information on the energy and the polarization of electronic transitions. These are valuable indicators for the evolution of electronic excitations in complex systems, where the electronic coupling between chromophores plays a key role. Highlighting collective effects that originate from electronic interactions in DNA multimers is the objective of the present Account.In contrast to the monomeric chromophores, whose fluorescence decays within a few picoseconds, that of DNA multimers persists on the nanosecond timescale. Even if long-lived states represent only a small fraction of electronic excitations, they may be crucial to the DNA photoreactivity because the probability to reach reactive conformations increases over time, owing to the incessant structural dynamics of nucleic acids.Our femtosecond studies have revealed that an ultrafast excitation energy transfer takes place among the nucleobases within duplexes and G-quadruplexes. Such an ultrafast process is possible when collective states are populated directly upon photon absorption. At much longer times, we discovered an unexpected long-lived high-energy emission stemming from what was coined "HELM excitons". These collective states, whose emission increases with the duplex size, could be responsible for the delayed fluorescence of ππ* states observed for genomic DNA.Most studies dealing with excited-state relaxation in DNA were carried out with excitation in the absorption band peaking at around 260 nm. We went beyond this and also performed the first time-resolved study with excitation in the UVA spectral range, where a very weak absorption tail is present. The resulting fluorescence decays are much slower and the fluorescence quantum yields are much higher than for UVC excitation. We showed that the base pairing of DNA strands enhances the UVA fluorescence and, in parallel, increases the photoreactivity because it modifies the nature of the involved collective excited states.

In higher-plant Photosystem I (PSI), the majority of "red" chlorophylls (absorbing at longer wavelengths than the reaction centre P 700 ) are located in the peripheral antenna, but contradicting reports are given about red forms in the core complex. Here we attempt to clarify the spectroscopic characteristics and quantify the red forms in the PSI core complex, which have profound implication on understanding the energy transfer and charge separation dynamics. To this end we compare the steady-state absorption and fluorescence spectra and picosecond time-resolved fluorescence kinetics of isolated PSI core complex and PSI-LHCI supercomplex from Pisum sativum recorded at 77 K. Gaussian decomposition of the absorption spectra revealed a broad band at 705 nm in the core complex with an oscillator strength of three chlorophylls. Additional absorption at 703 nm and 711 nm in PSI-LHCI indicated up to five red chlorophylls in the peripheral antenna. Analysis of fluorescence emission spectra resolved states emitting at 705, 715 and 722 nm in the core and additional states around 705-710 nm and 733 nm in PSI-LHCI. The red states compete with P 700 in trapping excitations in the bulk antenna, which occurs on a timescale of ~20 ps. The three red forms in the core have distinct decay kinetics, probably in part determined by the rate of quenching by the oxidized P 700 . These results affirm that the red chlorophylls in the core complex must not be neglected when interpreting kinetic experimental results of PSI., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 The Author(s). Published by Elsevier B.V. All rights reserved.)

A general theory is developed to identify the influence of local dipole fields on fluorescence and intermolecular electronic excitation transfer. The analysis, based on electrodynamical principles, identifies the fundamental quantum mechanisms and delivers full analytical results. The aim is to afford new physical insights, assisting the interpretation of measurements on the specific effects of local molecular dipoles on direct fluorescence and on fluorescence resonance energy transfer. Dipole field effects, which include those originating from intrinsically polar chromophores and surface field gradients, also prove to be manifest in electronic transitions of quadrupole symmetry character. The results have particular significance for fluorescence studies of cell membrane biophysics.

The photosynthetic Reaction Center (RC) from the purple bacterium Rhodobacter sphaeroides has unique photoconversion capabilities, that can be exploited in assembly biohybrid devices for applications in solar energy conversion. Extending the absorption cross section of isolated RC through covalent functionalization with ad-hoc synthesized artificial antennas is a successful strategy to outperform the efficiency of the pristine photoenzyme under visible light excitation. Here we report a new heptamethine cyanine antenna that, upon covalent binding to RC, forms a biohybrid (hCyN7-RC) which, under white light excitation, has doubled photoconversion efficiency versus the bare photoenzyme. The artificial antenna hCyN7 successfully meets appropriate optical properties, i.e. peak position of absorption and emission maximum in the visible and NIR region respectively, large Stokes shift, and high fluorescence quantum yield, required for improving the efficiency of the biohybrid in the production of the charge-separated state in the RC. The kinetics of energy transfer and charge separation of hCyN7-RC studied via ultrafast visible and IR spectroscopies are here presented. The antenna transfers energy to RC chromophores within <10 ps and the rate of Q A reduction is doubled compared to the native RC. These experiments further demonstrate hCyN7-RC, besides being an extremely efficient white light photoconverter, fully retains the charge separation mechanism and integrity of the native RC photoenzyme, thus allowing to envisage its suitability as biohybrid material in bioinspired systems for solar energy conversion., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Surface modified colloidal gold (Au) and silver (Ag) nanoparticles (NPs) were used as efficient quenchers of luminol (LH 2 ) fluorescence either in homogeneous aqueous medium or its noncovalent assembly with bovine serum albumin (BSA). The mechanism as well as the extent of fluorescence quenching was found to be strongly dependent on the nature of the nanoparticles. While simple static type fluorescence quenching mechanism was perceived with AuNP, a more complex protocol involving quenching sphere model was envisaged for AgNP quenching. Nevertheless, the magnitude of Stern-Volmer (SV) quenching constant (K SV ~ 10 8 -10 10  M -1 ) was calculated to be ca. 10 4 times more for surface quoted NPs in comparison with BSA-NP bioconjugates system. On the other hand, a highly efficient (E ≈ 95%) energy transfer (ET) process was predicted for LH 2 captured in the hydrophobic assembly with BSA in presence of AgNP as an acceptor. The ET efficiency is critically dependent on the concentration of BSA and nicely correlated with the extent of NP surface coverage. However, fluorescence quenching on AuNP surface is relatively less responsive towards protein concentration, primarily due to the difference in surface activity as well as the mode of interaction of the protein with NPs. Graphical Abstract Energy transfer from excited luminol to metal nanoparticles is strongly modulated in presence of serum albumins.

Photosynthetic organisms are frequently exposed to light intensities that surpass the photosynthetic electron transport capacity. Under these conditions, the excess absorbed energy can be transferred from excited chlorophyll in the triplet state (3Chl*) to molecular O 2 , which leads to the production of harmful reactive oxygen species. To avoid this photooxidative stress, photosynthetic organisms must respond to excess light. In the green alga Chlamydomonas reinhardtii , the fastest response to high light is nonphotochemical quenching, a process that allows safe dissipation of the excess energy as heat. The two proteins, UV-inducible LHCSR1 and blue light-inducible LHCSR3, appear to be responsible for this function. While the LHCSR3 protein has been intensively studied, the role of LHCSR1 has been only partially elucidated. To investigate the molecular functions of LHCSR1 in C. reinhardtii , we performed biochemical and spectroscopic experiments and found that the protein mediates excitation energy transfer from light-harvesting complexes for Photosystem II (LHCII) to Photosystem I (PSI), rather than Photosystem II, at a low pH. This altered excitation transfer allows remarkable fluorescence quenching under high light. Our findings suggest that there is a PSI-dependent photoprotection mechanism that is facilitated by LHCSR1., Competing Interests: The authors declare no conflict of interest.

Monitoring bacterial growth is an important technique required for many applications such as testing bacteria against compounds (e.g. drugs), evaluating bacterial composition in the environment (e.g. sewage and wastewater or food suspensions) and testing engineered bacteria for various functions (e.g. cellulose degradation). T?=1,^FigItem(1) ^ReloadFigure=Yesraditionally, rapid estimation of bacterial growth is performed using spectrophotometric measurement at 600nm (OD600) but this estimation does not differentiate live and dead cells or other debris. Colony counting enumerates live cells but the process is laborious and not suitable for large numbers of samples. Enumeration of live bacteria by flow cytometry is a more suitable rapid method with the use of dual staining with SYBR I Green nucleic acid gel stain and Propidium Iodide (SYBR-I/PI). Flow cytometry equipment and maintenance costs however are relatively high and this technique is unavailable in many laboratories that may require a rapid method for evaluating bacteria growth. We therefore sought to adapt and evaluate the SYBR-I/PI technique of enumerating live bacterial cells for a cheaper platform, a fluorimeter. The fluorimetry adapted SYBR-I/PI enumeration of bacteria in turbid growth media had direct correlations with OD600 (p>0.001). To enable comparison of fluorescence results across labs and instruments, a fluorescence intensity standard unit, the equivalent fluorescent DNA (EFD) was proposed, evaluated and found useful. The technique was further evaluated for its usefulness in enumerating bacteria in turbid media containing insoluble particles. Reproducible results were obtained which OD600 could not give. An alternative method based on the assessment of total protein using the Pierce Coomassie Plus (Bradford) Assay was also evaluated and compared. In all, the SYBR-I/PI method was found to be the quickest and most reliable. The protocol is potentially useful for high-throughput applications such as monitoring of growth of live bacterial cells in 96-well microplates and in assessing in vivo activity of cellulose degrading enzyme systems., (Copyright © 2017 Elsevier B.V. All rights reserved.)

DNA crystals make it possible to organize guest molecules into specific periodic 3D patterns at the nanoscale, and thereby to create novel macroscopic objects with potentially useful functionality. Here, we describe the fluorescence and energy transfer properties of DNA crystals that are self-assembled from DNA tensegrity triangles with covalently attached Cy3 and Cy5 dyes. When compared to reference DNA strands in solution, the fluorescence measurements indicate that the dyes in the crystal experience a more homogeneous environment, resulting in a 2-fold increase in Cy3 quantum yield and single-exponential Cy3 fluorescence decays. Energy transfer in a network of coupled Cy3 and Cy5 dyes in the DNA crystal is demonstrated experimentally. Numerical simulation finds the experiments to be consistent with a Förster model of the dyes in the periodic crystalline environment, and particularly if the transition dipoles are assumed random in orientation but static on the time scale of the excitation decay.

Purpose: The authors describe a method in which fluorescence nuclear track detectors (FNTDs), novel track detectors with nanoscale spatial resolution, are used to determine the linear energy transfer (LET) of individual proton tracks from proton therapy beams by allowing visualization and 3D reconstruction of such tracks., Methods: FNTDs were exposed to proton therapy beams with nominal energies ranging from 100 to 250 MeV. Proton track images were then recorded by confocal microscopy of the FNTDs. Proton tracks in the FNTD images were fit by using a Gaussian function to extract fluorescence amplitudes. Histograms of fluorescence amplitudes were then compared with LET spectra., Results: The authors successfully used FNTDs to register individual proton tracks from high-energy proton therapy beams, allowing reconstruction of 3D images of proton tracks along with delta rays. The track amplitudes from FNTDs could be used to parameterize LET spectra, allowing the LET of individual proton tracks from therapeutic proton beams to be determined., Conclusions: FNTDs can be used to directly visualize proton tracks and their delta rays at the nanoscale level. Because the track intensities in the FNTDs correlate with LET, they could be used further to measure LET of individual proton tracks. This method may be useful for measuring nanoscale radiation quantities and for measuring the LET of individual proton tracks in radiation biology experiments.

Monitoring the interaction of biomolecules is important, and the use of energy transfer is a principal technique in elucidating nanoscale interactions. Lanthanide compounds are promising luminescent probes for biological samples as their emission is longer-lived than any native autofluorescence. Polyoxometalates (POMs) are interesting structural motifs to incorporate lanthanides, offering low toxicity and a size pertinent for biological applications. Here, we employ iso-structured POMs containing either terbium or europium and assess their interaction with serum albumin by sensitisation of a fluorescent tag on the protein via LRET (luminescence resonance energy transfer) by exciting the lanthanide. Time-resolved measurements showed energy transfer with an efficiency of over 90% for the POM-protein systems. The Tb-POM results were relatively straightforward, while those with the iso-structured Eu-POM were complicated by the effect of protein shielding from the aqueous environment., (© 2016 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A NaYF4 host matrix with a new cubic phase is fabricated to offer high upconversion luminescence efficiency. The new cubic phase is formed through a hexagonal-to-cubic phase transition by shining intense near-infrared light on lanthanide-doped hexagonal NaYF4 materials., (© 2015 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

A strategy to probe supramolecular nanocarriers and their cargo in the intracellular space was developed on the basis of fluorescence measurements and energy transfer. It relies on the covalent attachment of an energy donor, or acceptor, to the macromolecular backbone of amphiphilic polymers and the noncovalent encapsulation of a complementary acceptor, or donor, in the resulting micelles. In aqueous environments, these macromolecules self-assemble into nanostructured constructs and bring the complementary chromophores in close proximity to enable efficient energy transfer. These supramolecular assemblies travel from the extracellular to the intracellular space and retain their integrity in the process. Indeed, donors and acceptors remain close to each other after internalization, and excitation of the former chromophores translates into significant intracellular emission from the latter. Furthermore, these supramolecular assemblies exchange their components with fast kinetics in aqueous dispersions because of the reversible character of the noncovalent contacts holding them together. As a result, micelles incorporating exclusively the donors and nanocarriers containing only the acceptors scramble their chromophoric building blocks, upon mixing, to allow the transfer of energy. These dynamic processes can be reproduced in the intracellular environment with the sequential incubation of cells with the two sets of complementary nanostructured assemblies. Thus, these operating principles and choice of supramolecular synthons are particularly valuable to monitor self-assembling nanocarriers and their cargo inside living cells and can facilitate the elucidation of the behavior of these promising delivery vehicles in a diversity of biological specimens.

We report on theoretical efficiency of non-photochemical fluorescense quenching of phycobilisomes by the orange carotenoid protein. The created 3D computer model of the three-cylindrical phycobilisomes core allowed us to determine the distances between centers of mass of all phycobilin chromophores of the core and calculate the time and an average number of energy migration steps for the resulting non-radiative excitation transfer from the phycobilisomes to photosystem II. The obtained kinetic scheme equations for a way of energy transfer confirm the incomplete interception of energy flow in the phycobilisomes core by the orange carotenoid protein. Theoretical estimation of the rate of phycobilisomes quenching is in good agreement with experimental data.

The results of simulations of fluorescence line narrowed (FLN) and Δ-FLN spectra of weakly coupled chromophore dimers and trimers, connected by slow (hundreds of picoseconds) excitation energy transfer and embedded in amorphous solids such as proteins, are presented. The model includes wavelength-dependent excitation energy transfer (EET) rate distributions converted to nonidentical sub- site distribution function (sub-SDF) (one for each EET rate), as well as spectral hole burning in pigments excited via EET. Emission of the donor molecules and realistic EET probability dependence on the donor-acceptor energy gap are also included. The shapes of the FLN and Δ-FLN spectra are very sensitive to even small variations of interpigment coupling in this weak-coupling regime, and proper modeling is essential for extracting correct parameters from the experimental spectra. Applications of the model to the trimeric FMO complex and the dimeric cytochrome b6f are demonstrated.

Based on a through bond energy transfer (TBET) between rhodamine and naphthalimide fluorophores, a fluorescent ratiometric chemosensor L was designed and prepared for highly selective detection of Fe(3+) in aqueous solution and in living EC109 cells. These significant changes in the fluorescence color could be used for naked-eye detection. The reversibility established the potential of the probe as chemosensor for Fe(3+) detection., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The plant light-harvesting complexes of photosystem II (LHC-II) play important roles in collecting solar energy and transferring the energy to the reaction centers of photosystems I and II. A two photon absorption compound, 4-(bromomethyl)-N-(4-(dimesitylboryl)phenyl)-N-phenylaniline (DMDP-CH2Br), was synthesized and covalently linked to the LHC-II in formation of a LHC-II-dye complex, which still maintained the biological activity of LHC-II system. Under irradiation with femtosecond laser pulses at 754 nm, the LHC-II-dye complex can absorb two photons of the laser light effectively compared with the wild type LHC-II. The absorbed excitation energy is then transferred to chlorophyll a with an obvious fluorescence enhancement. The results may be interesting and give potentials for developing hybrid photosystems., (Copyright © 2014 Elsevier B.V. All rights reserved.)

The deterministic identifiability analysis of photophysical models for the kinetics of excited-state processes, assuming errorless time-resolved fluorescence data, can verify whether the model parameters can be determined unambiguously. In this work, we have investigated the identifiability of several uncommon models for time-resolved fluorescence with underlying distributions of rate constants which lead to non-exponential decays. The mathematical functions used here for the description of non-exponential fluorescence decays are the stretched exponential or Kohlrausch function, the Becquerel function, the Förster type energy transfer function, decay functions associated with exponential, Gaussian and uniform distributions of rate constants, a decay function with extreme sub-exponential behavior, the Mittag-Leffler function and Heaviside's function. It is shown that all the models are uniquely identifiable, which means that for each specific model there exists a single parameter set that describes its associated fluorescence δ-response function.

Photochromic 1,2-dithienylethene (DTE) derivatives with a high thermal stability and fatigue resistance are appealing for optical switching of fluorescence. Here, we introduce a donor-photochromic bridge-acceptor tetraphenylethene-dithienylethene-perylenemonoimide (TPE-DTE-PMI) triad, in which TPE acts as the electron donor, PMI as the electron acceptor, and DTE as the photochromic bridge. In this system, the localized and intramolecular charge transfer emission of TPE-DTE-PMI with various Stokes shifts have been observed due to the photoinduced intramolecular charge transfer in different solvents. Upon UV irradiation, the fluorescence quenching resulting from photochromic fluorescence resonance energy transfer in TPE-DTE-PMI has been demonstrated in solution and in solid films. The fluorescence on/off switching ratio in polymethylacrylate film exceeds 100, a value much higher than in polymethylmethacrylate film, thus indicating that the fluorescence switching is dependent on matrices., (Copyright © 2014 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

Principles behind quenching of tryptophan (Trp) fluorescence are updated and extended in light of recent 100-ns and 1-μs molecular dynamics (MD) trajectories augmented with quantum mechanical (QM) calculations that consider electrostatic contributions to wavelength shifts and quenching. Four studies are summarized, including (1) new insight into the single exponential decay of NATA, (2) a study revealing how unsuspected rotamer transitions affect quenching of Trp when used as a probe of protein folding, (3) advances in understanding the origin of nonexponential decay from 100-ns simulations on 19 Trps in 16 proteins, and (4) the correlation of wavelength with lifetime for decay-associated spectra (DAS). Each study strongly reinforces the concept that-for Trp-electron transfer-based quenching is controlled much more by environment electrostatic factors affecting the charge transfer (CT) state energy than by distance dependence of electronic coupling. In each case, water plays a large role in unexpected ways.

A full-color molecular pixel system is realized for the first time using simple mixtures composed of RGB-emitting excited-state intramolecular proton transfer (ESIPT) dyes, each of which has delicately tailored Stokes shift and independent emission capability completely free from energy transfer crosstalk between them. It is demonstrated that the whole range of emission colors enclosed within the RGB color triangle on the CIE 1931 diagram is predictable and conveniently reproducible from the RGB molecular pixels not only in the solution but also in the polymer film. It must be noted that mixing ratios to reproduce the desired color coordinates can be precisely calculated on the basis of additive color theory according to their molecular pixel behavior.

We report phosphorescent sensitized fluorescent near-infrared (NIR) light-emitting electrochemical cells (LECs) utilizing a phosphorescent cationic transition metal complex [Ir(ppy)(2)(dasb)](+)(PF(6)(-)) (where ppy is 2-phenylpyridine and dasb is 4,5-diaza-9,9'-spirobifluorene) as the host and two fluorescent ionic NIR emitting dyes 3,3'-diethyl-2,2'-oxathiacarbocyanine iodide (DOTCI) and 3,3'-diethylthiatricarbocyanine iodide (DTTCI) as the guests. Photoluminescence measurements show that the host-guest films containing low guest concentrations effectively quench host emission due to efficient host-guest energy transfer. Electroluminescence (EL) measurements reveal that the EL spectra of the NIR LECs doped with DOTCI and DTTCI center at ca. 730 and 810 nm, respectively. Moreover, the DOTCI and DTTCI doped NIR LECs achieve peak EQE (power efficiency) up to 0.80% (5.65 mW W(-1)) and 1.24% (7.84 mW W(-1)), respectively. The device efficiencies achieved are among the highest reported for NIR LECs and thus confirm that phosphorescent sensitized fluorescence is useful for achieving efficient NIR LECs., (This journal is © the Owner Societies 2011)

The mutual interaction of imidazole derivative (PIPP) with bovine serum albumin (BSA) was investigated using photoluminescent studies. The fluorescence quenching mechanism of BSA by PIPP was analyzed and the binding constant was calculated. The binding distance between PIPP and BSA was obtained based on the theory of Forester's non-radiation energy transfer. Displacement experiments were performed by using ibuprofen to identify PIPP binding site in BSA. The effect of some common ions on the binding constant between PIPP and BSA was also examined., (© Springer Science+Business Media, LLC 2011)

The interaction between quercetin and lipoxygenase was investigated by fluorescence spectroscopy. The analysis of the emission quenching at different temperatures revealed that the quenching mechanism correspond to a static process and, as consequence, a complex quercetin-lipoxygenase is formed. The thermodynamic parameters ΔG, ΔH and ΔS were calculated to be-32.57 kJ mol(-1),-3.21 kJ mol(-1) and 87.14 J mol(-1) K(-1) respectively, which suggest that hydrophobic forces plays a major role in the stabilization of the complex quercetin-lipoxygenase. The distance, r, between donor (lipoxygenase) and acceptor (quercetin) was calculated to be 3.84 nm based on Förster's non-radiative energy transfer theory. The results obtained from the evaluation of three dimensional florescence spectra suggest a conformational modification of the protein in the region of the coupling with quercetin., (© Springer Science+Business Media, LLC 2011)

Fluorescence intensity correlation functions contain information about photophysical and conformational dynamics. We propose and implement a simple procedure to analyze such functions measured in the presence of resonance energy transfer. When there is a separation of time scales and the conformational dynamics is modeled as diffusion in the potential of mean force along the interdye distance, we obtain an analytic expression for the conformational correlation time. This can be used to find the diffusion coefficient describing conformational fluctuations given the photon count rate and equilibrium distribution.

Energy transfer between photosystem II (PSII) centers is known from previous fluorescence studies. We have studied the theoretical consequences of energetic connectivity of PSII centers on photosynthetic thermoluminescence (TL) and predict that connectivity affects the TL Q band. First, connectivity is expected to make the Q band wider and more symmetric than an ideal first-order TL band. Second, the presence of closed PSII centers in an energetically connected group of PSII centers is expected to lower the probability that an exciton originating in a recombination reaction becomes retrapped. The latter effect would shift the Q band toward lower temperature, and the shift would be greater the higher the percentage of closed PSII centers at the beginning of the measurement. These effects can be generalized as second-order effects, as they make the Q band resemble the second-order TL bands obtained from semiconducting solids. We applied the connected-units model of chlorophyll fluorescence to derive equations for quantifying the second-order effects in TL. To test the effect of the initial proportion of closed reaction centers, we measured the Q band with different intensities of the excitation flash and found that the peak position changed by 2.5 degrees C toward higher temperature when the flash intensity was lowered from saturating to 0.39% of saturating. The result shows that energy transfer between reaction centers of PSII forms the physical basis of retrapping in photosynthetic TL. The second-order effects partially explain the deviation of the form of the Q band from ideal first-order TL.

An investigation of the mechanism of the fluorescence quenching by Cu(2+) for a conjugated polymer system initially designed as a fluorescence "turn-on" chemosensor based on chelation enhanced fluorescence (CHEF) is described in this paper. Unlike all other metal cations tested, the polymer/Cu(2+) hybrid system with a 1:1 ratio between the receptor and Cu(2+) has only weak fluorescence with lambda(max) = 490 nm and a quantum yield of 0.004 in THF at room temperature. In solvent glasses at 77 K the fluorescence remained quenched suggesting that the quenching mechanism was due to energy transfer between the Cu(2+) and the conjugated polymer backbone. The energy transfer quenching competes effectively with the electron transfer involved in the CHEF resulting in a more selective chemosensory system.

Interaction of 3-styrylindoles 1-8 viz. 3-(2-phenylethenyl-E)-NH-indole (1), 3-[2-(4-nitrophenyl)ethenyl-E]-NH-indole (2), 5-bromo-3-[2-(4-nitrophenyl)ethenyl-E]-NH-indole (3), 5-methoxy-3-[2-(4-nitrophenyl)ethenyl-E]-NH-indole (4), 3-[2-(4-cyanophenyl)ethenyl-E]-NH-indole (5), 3-[2-(4-cyanophenyl)ethenyl-E]-N-ethylindole (6), 5-bromo-3-[2-(4-chlorophenyl)ethenyl-E]-NH-indole (7) and 5-methoxy-3-[2-(4-chlorophenyl)ethenyl-E]-NH-indole (8) with bovine serum albumin (BSA) was examined by UV-vis and steady-state fluorescence spectroscopy. The fluorescence intensity of 1-8 increases with the increasing BSA concentration. Upon binding with BSA, while 1 and 5-8 show a blue shift in their lambda(f max), 2-4 do not exhibit such behavior. Compounds 1-8 also quench the 345 nm fluorescence of BSA in phosphate buffer (lambda(ex), 280 nm). These compounds intercalate in the hydrophobic regions of BSA, as evidenced by the determination of BSA binding site micropolarity using compounds 2-8. As evidenced by the estimation of energy transfer efficiency and distance between the donor (BSA-Trp-212) and the acceptor (3-styrylindoles), the halo-substituted compounds 3 and 7 interact with BSA more effectively than the other 3-strylindoles. These compounds have potential for use as neutral and hydrophobic fluorescence probes for examining the microenvironments in proteins, polymers, micelles, etc.

A diaryldiketopyrrolopyrrole derivative functionalized with phenothiazine moieties (DPPP) was synthesized and introduced into the ordered 4-(3,6-di-tert-butyl-9H-carbazol-9-yl)benzamide (TBCB) organogel system. It was found that TBCB-based gel became a scaffold to make DPPP molecules line up along the gel fibers, resulting in new self-assembled arrays, whose XRD patterns were quite different from those of the neat TBCB gel and DPPP crystal. In the composite gel, the occurrence of a partial energy transfer from the excited light-harvesting antenna of TBCB to the DPPP acceptor was confirmed on the basis of time-dependent and time-resolved fluorescence investigations. Remarkably, the composite gel could emit intense red light or purplish white light by tuning the excitation wavelength. Such ordered soft materials with color-tunable emission may possess potential applications in sensor and photonic devices.

In order to survive sunlight in the absence of water, desiccation-tolerant green plants need to be protected against photooxidation. During drying of the chlorolichen Cladonia rangiformis and the cyanolichen Peltigera neckeri, chlorophyll fluorescence decreased and stable light-dependent charge separation in reaction centers of the photosynthetic apparatus was lost. The presence of light during desiccation increased loss of fluorescence in the chlorolichen more than that in the cyanolichen. Heating of desiccated Cladonia thalli, but not of Peltigera thalli, increased fluorescence emission more after the lichen had been dried in the light than after drying in darkness. Activation of zeaxanthin-dependent energy dissipation by protonation of the PsbS protein of thylakoid membranes was not responsible for the increased loss of chlorophyll fluorescence by the chlorolichen during drying in the light. Glutaraldehyde inhibited loss of chlorophyll fluorescence during drying. Desiccation-induced loss of chlorophyll fluorescence and of light-dependent charge separation are interpreted to indicate activation of a highly effective mechanism of photoprotection in the lichens. Activation is based on desiccation-induced conformational changes of a pigment-protein complex. Absorbed light energy is converted into heat within a picosecond or femtosecond time domain. When present during desiccation, light interacts with the structural changes of the protein providing increased photoprotection. Energy dissipation is inactivated and structural changes are reversed when water becomes available again. Reversibility of ultra-fast thermal dissipation of light energy avoids photo-damage in the absence of water and facilitates the use of light for photosynthesis almost as soon as water becomes available.

The origination of the peak at 730 nm in the delayed fluorescence (DF) spectrum of chloroplasts was studied using various optical analysis methods. The DF spectrum showed that the main emission peak was at about 685 nm, with a small shoulder at 730 nm when the chloroplast concentration was < 7.8 microg/mL. The intensity of the peak at 685 nm decreased, while the intensity of the peak at 730 nm increased, when the chloroplast concentrations were increased from 7.8 to 31.2 microg/mL. With the concentration increasing, the peak at 730 nm became dominant while the peak at 685 nm finally disappeared. The DF decay kinetic curves showed that the intensity of the peak at 730 nm decayed as the same speed as the intensity of the peak at 685 nm during the entire relaxation process (0.5-30.5 s). With the excitation wavelength at 685 nm, the emission intensity was stronger in the excitation spectrum at 730 nm. The absorption spectrum demonstrated that the ratio A(685):A(730) remained almost constant when the chloroplast concentration increased. The results suggest that the peak at 730 nm appearing in DF is mainly contributed by the fluorescence of photosystem I (PSI), generated by the re-absorption of 685 nm band DF., ((c) 2007 John Wiley & Sons, Ltd.)

Necessary modifications to the expression for the Förster energy transfer rate are discussed when fluorescence decay of the donor in the absence of acceptor is nonexponential. Discrete and continuous models of the nonexponentiality are taken into account. No general solution of the problem is found. It is, however, suggested that in many of the biochemical problems the most appropriate modification of the transfer rate can be that which is based on the assumption of the same constant value of the radiative decay rate for all donor molecules. The effect of the assumed form of the Förster energy transfer rate on the recovered values of the distance distribution and dynamics parameters of some exemplary bichromophoric systems is examined.

Excitation energy transfer in DNA has similarities to charge transfer, but the transport is of an excited state, not of mass or charge. Use of the fluorescent, modified adenine base 2-aminopurine (2AP) as an energy trap in short (3- to 20-base) single- and double-stranded DNA oligomers is reviewed. Variation of 2AP's neighboring sequence shows (1) relatively efficient transfer from adenine compared to that from cytosine and thymine, (2) efficient transfer from guanine, but only when 2AP is at the 3' end, (3) approximate equality of efficiencies for 3' to 5' and 5' to 3' directional transfer in adenine tracks. The overall, average transfer distance at room temperature is about four adenine bases or less before de-excitation. The transfer fluorescence excitation spectral shape is similar to that of the absorption spectrum of the neighboring normal bases, confirming that initial excitation of the normal bases, followed by emission from 2AP (i.e. energy transfer), is occurring. Transfer apparently may take place both along one strand and cross-strand, depending on the oligomer sequence. Efficiency increases when the temperature is decreased, rising above 50% (overall efficiency) in decamers of adenine below -60 degrees C (frozen media). Modeling of the efficiencies of transfer from the nearest several adenine neighbors of 2AP in these oligomers suggests that the nearest two neighbors transfer with near 100% efficiency. As bases in B DNA, as well as in single-stranded DNA, are separated by less than 5 A (less than the size of a base), standard Förster transfer theory should not apply. Indeed, while both theory and experiment show efficiency decreasing with donor-acceptor distance, the experimental dependence clearly disagrees with Förster 1/r6 dependence. It is not yet clear what the best theoretical approach is, but any calculation must deal accurately with the excited states of bases, including strong base-base interactions and structural fluctuations, and should reflect the increase of efficiency with temperature decrease and the relative insensitivity to strandedness (single, double). Attempts to use DNA as a molecular "fiber optic" face three primary challenges. First, reasonable efficiency over more than a base or two occurs only in adenine stretches at temperatures well below freezing. Second, transfer in these adenine tracks is efficient in both directions. Third, absorption of UV light occurs randomly, making excitation at a specific site on this "fiber optic" a challenge.

We report on 2.0-microm emission characteristic and energy transfer of Ho3+-doped tellurite glass sensitized by Tm3+ upon excitation of a conventional 808 nm laser diode. The Judd-Ofelt strength parameters, spontaneous radiative transition probabilities and radiative lifetime of Ho3+ have been calculated from the absorption spectra by using the Judd-Ofelt theory. Significant enhancement of 2.0-microm emission of Ho3+ has been observed with increasing Tm3+ doping up to 0.7 mol%. The energy transfer coefficient of the forward Tm3+-->Ho3+ is approximately 17 times larger than that of the backward Tm3+<--Ho3+ energy transfer. Our result indicates that the maximum gain of 2.0-microm emission, assigned to the transition of (5)I7-->(5)I8 of Ho3+, might be achieved from the tellurite glass at the concentration of 0.5 mol% of Tm2O3 and 0.15 mol% of Ho2O3. The high gain coefficient and quantum efficiency (1.16) along with the large value of the product of the stimulated emission cross-section and the measured radiative lifetime (4.12 x 10(-27) m(2)s) of the Ho3+/Tm3+-codoped tellurite glasses might find potential applications in efficient 2.0-microm laser.

Next-generation photodynamic therapy agents will minimize extraneous phototoxicity by being active only at the target site. To this end, we have developed a model system to systematically investigate the excited-state quenching ability of a number of metalloporphyrins. Central metal ions that prefer four-coordinate, square planar orientations (Ag(II), Cu(II), Ni(II), Pd(II), and Zn(II)) were used. Porphyrin dimers based on 5-(4-aminophenyl)-10,15,20-triphenylporphyrin and comprising both a free base porphyrin and a metalloporphyrin covalently linked through a five-carbon alkyl chain were synthesized. The fluorescence and singlet oxygen quantum yields for the dimers were probed at 630 and 650 nm, respectively, resulting in the excitation of only the free base porphyrin and allowing a comparison of the quenching efficacy of each central metal ion. These results demonstrate that metalloporphyrins can serve as efficient quenchers, and may be useful in the design of novel light-activated therapeutic agents.

A sensitive fluorescence enhancement system was developed for the determination of terbium. The fluorescence intensity of the Tb-N-(2-pyridinyl) ketoacetamide (PKAA) system was greatly enhanced by the addition of triethylamine (Et(3)N) and zinc nitrate in the methanol solution. The excitation and emission wavelengths were 329 and 546 nm, respectively. Under optimal conditions, the fluorescence intensities varied linearly with the concentration of Tb(3+) in the range of 7.5 x 10(-8)-8.2 x 10(-6)M with a detection limit of 6.4 x 10(-8)M. The interferences of some rare earth ions were described. This method was applied to the determination of trace amounts of terbium (III) in a synthetic rare earth oxide and a high purity Y(2)O(3) matrix. The mechanism of fluorescence enhancement was also studied.

In the present paper, a fluorescence method was used to study at different pH the fluorescence quenching of bovine serum albumin (BSA) by its interaction with balofloxacin (BLFX). The interaction association constants of BSA and BLFX were determined from a double reciprocal line Weaver-Burk plot. According to the Forster dipole-dipole energy transfer, the distance to be measured between the BLFX and tryptophane is 5.09 nm. From thermodynamical coordination it can be judged that the binding power between BLFX and BSA is electrostatic effect. The effect of BLFX on the conformation of BSA was also analyzed by using synchronous fluorescence spectroscopy.

The use of two-photon excitation of fluorescence for detection of fluorescence resonance energy transfer (FRET) was studied for a selected fluorescent donor-acceptor pair. A method based on labeled DNA was developed for controlling the distance between the donor and the acceptor molecules. The method consists of hybridization of fluorescent oligonucleotides to a complementary single-stranded target DNA. As the efficiency of FRET is strongly distance dependent, energy transfer does not occur unless the fluorescent oligonucleotides and the target DNA are hybridized. A high degree of DNA hybridization and an excellent FRET efficiency were verified with one-photon excited fluorescence studies. Excitation spectra of fluorophores are usually wider in case of two-photon excitation than in the case of one-photon excitation. This makes the selective excitation of donor difficult and might cause errors in detection of FRET with two-photon excited fluorescence. Different techniques to analyze the FRET efficiency from two-photon excited fluorescence data are discussed. The quenching of the donor fluorescence intensity turned to be the most consistent way to detect the FRET efficiency. The two-photon excited FRET is shown to give a good response to the distance between the donor and the acceptor molecules.

The spectral evolution of three photoactive proteins has been investigated by measuring the fluorescence with good temporal and wavelength resolution and a high signal-to-noise ratio. Upon excitation at 400 nm wild-type (wt) PYP both at neutral pH and in the low-pH blueshifted pBdark state exhibited a strong quenching of the fluorescence, the major part of which could be described by lifetimes of about 1.7 and 7.7 ps. The remaining fluorescence decay occurred multiexponentially with lifetimes between 30 and 125 ps. Additionally, in wtPYP at neutral pH, a dynamic Stokes shift was found to occur with a time constant of about 0.25 ps. In a PYP preparation that was reconstituted with the chromophore 7-hydroxy-coumarin-3- carboxylic acid rather than the native coumaric acid, and which is therefore not capable of performing the cis-trans-isomerization that initiates the photocycle in wtPYP, the fluorescence was found to decay multiexponentially with lifetimes of 51 ps, 0.33 and 3.77 ns. Additionally, dynamic Stokes shifts were observed with time constants of about 0.1 and 3.5 ps. Upon comparison of the dynamics of this preparation with that of wtPYP the multiexponential decay with lifetimes of 1.7 and 7.7 ps found in wtPYP was attributed to photochemistry of the p-coumaric-acid chromophore. The emission from bacteriorhodopsin mutant D85S upon excitation at 635 nm decays biexponentially with estimated lifetimes of 5.2 and 19.1 ps. No dynamic Stokes shift was observed here. Four lifetimes were needed to describe the decay of the emission from the A* state in the green fluorescent protein. From a target analysis it was concluded that the longer lifetimes are accompanied by a decreasing probability of forming I*, which approaches zero with the longest A* lifetime of 1.5 ns. These observations may be explained by heterogeneity of A and by relaxation of A*. In all three systems studied, multiexponential decay of emission was present, suggesting that heterogeneity is a common feature of these chromophore protein complexes.

The interaction between carbamazepine (CBZ) and bovine serum albumin (BSA) was studied using fluorescence spectroscopy (FS) and ultraviolet spectroscopy (UV). The experimental results showed that the CBZ could insert into the BSA and quench the inner fluorescence of BSA by forming the CBZ-BSA complex. It was found that both static quenching and non-radiation energy transfer were the main reasons leading to the fluorescence quenching. The apparent binding constants (K) between CBZ and BSA were found to be 1.8 x 10(4) (27 degrees C) and 2.8 x 10(4) (37 degrees C) and the binding site values (n) were 0.97 (27 degrees C) and 1.01 (37 degrees C), respectively. According to the Forster theory of non-radiation energy transfer, the binding distances (r) between CBZ and BSA were 3.6 nm and 3.4 nm at 27 degrees C and 37 degrees C, respectively. The process of the binding was a spontaneous molecular interaction in which entropy increased and Gibbs free energy decreased, indicating that the interaction between CBZ and BSA was mainly driven by the hydrophobic force.

We show from time-resolved fluorescence intensity and depolarization experiments that the fluorescence of the unique tryptophan W119 of PYP is quenched by energy transfer to the 4-hydroxycinnamoyl chromophore. Whereas the intensity decay has a time constant of 0.18 ns in P, the decay in the absence of the cofactor (apo-PYP) has a single exponential lifetime of 4.8 ns. This difference in lifetime with and without acceptor can be explained quantitatively on the basis of energy transfer and the high-resolution X-ray structure of P, which allows an accurate calculation of the kappa2 factor. Fluorescence depolarization experiments with donor and acceptor indicate that both are immobilized so that kappa2 is constant on the fluorescence time scale. Using background illumination from an LED emitting at 470 nm, we measured the time-resolved fluorescence in a photostationary mixture of P and the intermediates I2 and I2'. The composition of the photostationary mixture depends on pH and changes from mainly I2 at low pH to predominantly I2' at high pH. The I2/I2' equilibrium is pH-dependent with a pKa of approximately 6.3. In I2 the lifetime increases to approximately 0.82 ns. This is not due to a change in distance or to the increase in spectral overlap but is primarily a consequence of a large decrease in kappa2. Kappa2 was calculated from the available X-ray structures and decreases from approximately 2.7 in P to 0.27 in I2. This change in kappa2 is caused by the isomerization of the acceptor, which leads to a reorientation of its transition dipole moment. We have here a rare case of the kappa2 factor dominating the change in energy transfer. The fluorescence decay in the light is pH-dependent. From an SVD analysis of the light/dark difference intensity decay at a number of pH values, we identify three species with associated lifetimes: P (0.18 ns), I2 (0.82 ns), and X (0.04 ns). On the basis of the pH dependence of the amplitudes associated with I2 and X, with a pKa of approximately 6.3, we assign the third species to the signaling state I2'. The absorption spectra of the 0.82 and 0.04 ns species were calculated from the pH dependence of their fluorescence amplitudes and of the photostationary light/dark difference absorption spectra. The lambda(max) values of these spectra (372 and 352 nm) identify the 0.82 and 0.04 ns components with I2 and I2', respectively, and validate the fluorescence data analysis. The mutant E46Q allows a further test of the energy transfer explanation, since lowering the pH in the dark leads to a bleached state with an increased spectral overlap but without the isomerization-induced decrease in kappa2. The measured lifetime of 0.04 ns is in excellent agreement with predictions based on energy transfer and the X-ray structure.

A new fluorochromic dye was obtained from the reaction of 9-aminoacridine with ethyl-2-cyano-3-ethoxyacrylate. It displays complex fluorescence that is ascribed to normal emission from the acridine chromophore in addition to excited-state intramolecular charge transfer (ESICT) formed upon light excitation. The analysis of the fluorescence decays in different solvents reveals two short-lived components in the range of 80-450 ps and 0.7-3.2 ns, ascribed to the formation and decay of the intramolecular charge transfer (ICT) state, in addition to a third component of about 9.0 ns, which is related to the normal emission from the acridine singlet excited state, probably in an enol-imine tautomeric form. The ICT emission is readily quenched by water addition to polar solvents, and this effect is ascribed to changes in the keto-amine/enol-imine equilibrium of this fluorochromic dye.

An energy transfer technique between acridine orange (AO) and rhodamine 6G (R6G) was studied, and the optimum experimental conditions of energy transfer were defined. It was found that the effective energy transfer could occur between AO and R6G in the dodecylbenzene sodium sulfonate solution with Na2 HPO4-citric acid buffer solution at pH 5.0. The fluorescence intensity of AO-R6G system was diminished by vitamin B12 in an alkalescence medium. Based on the AO-R6G energy transfer system anovel fluorescence quenching method for the determination of vitamin B12 has been developed. Under optimal conditious, the linear range of calibration curves for the determination of vitamin B12 was 0-3.0 x 10(-5) mol x L(-1). The detection limits were 4.8 x 10(-7) mol x L(-1) for Vitamin B12. Among six times of determination, the relative standard deviation was 0.51%-0.64%, and the recovery was 98.40% -103.62%. The method features good recurrence, rapidity of reaction, good stability, and few interfering substances. It can be satisfactorily used in the determination of the injection content of vitamin B12.

Low-temperature absorption and fluorescence spectra of fully active cores and membrane-bound PS II preparations are compared. Detailed temperature dependence of fluorescence spectra between 5 and 70 K are presented as well as 1.7-K fluorescence line-narrowed (FLN) spectra of cores, confirming that PS II emission is composite. Spectra are compared to those reported for LHCII, CP43, CP47 and D1/D2/cytit b559 subunits of PS II. A combination of subunit spectra cannot account for emission of active PS II. The complex temperature dependence of PS II fluorescence is interpretable by noting that excitation transfer from CP43 and CP47 to the reaction centre is slow, and strongly dependent on the precise energy at which a 'slow-transfer' pigment in CP43 or CP47 is located within its inhomogeneous distribution. PS II fluorescence arises from CP43 and CP47 'slow-transfer' states, convolved by this dependence. At higher temperatures, thermally activated excitation transfer to the PS II charge-separating system bypasses such bottlenecks. As the charge-separating state of active PS II absorbs at >700 nm, PS II emission in the 680-700 nm region is unlikely to arise from reaction centre pigments. PS II emission at physiological temperatures is discussed in terms of these results.

Delayed fluorescence dark decays in the time interval from 0.35 to 5.5 ms are measured during dark to light adaptation in whole barley leaves and isolated thylakoid membranes, using a disc phosphoroscope. The changes in delayed fluorescence features are compared with variable chlorophyll fluorescence simultaneously registered with the same apparatus as well as in parallel by Handy PEA (Hansatech Instruments Ltd.), and absorbance changes at 820 nm. The registered delayed fluorescence signal is a sum of three components - submillisecond with lifetime of about 0.6 ms, millisecond decayed 2-4 ms and slow component with lifetime > >5.5 ms. The submillisecond delayed fluorescence component is proposed to be a result of radiative charge recombination in Photosystem II reaction centers in the state Z+PQA -QB -, and its lifetime is determined by the rate of electron transfer from QA - to QB -. The millisecond delayed fluorescence component is associated with recombination in Z+PQA -QB = centers with a lifetime determined by the sum of the rate constants of electron transfer from the oxygen-evolving complex to Z+ and of the exchange between the reduced and oxidized plastoquinone pool in the QB-site. On the basis of these assumptions and of the different share of the three components in the integral delayed fluorescence during induction, an attempt has been made to interpret the changes in the delayed fluorescence intensity during the transition of the photosynthetic apparatus from dark to light adapted state.

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

The binding reaction of methyl pheophorbide-a with bovine serum albumins (BSA) in aqueous solution was studied by fluorescence and UV-Vis absorption spectra. The results indicated that the combination reaction of them was a single static quenching process. In aqueous solution, methyl pheophorbide-a strongly bound BSA with the apparent molar ratio of 2 : 1 because of methyl pheophorbide-a polymerized by itself. The binding constant KB was 6.7 x 10(4) L x mol(-1). In mixture solvent of tetrahydrofuran and water, methyl pheophorbide-a existed as single molecule and bound BSA with a molar ratio of 1: 1. There is single position for combining methyl pheophorbide-a with BSA. The shortest binding distance (r = 3.50 nm) and energy transfer efficiencies (E = 0.39) between donor (BSA) and acceptor (methyl pheophorbide-a) were obtained by Förster's nonradiative energy transfer mechanism.

Purpure-18-imide was synthesized based on the porphyrin's special affinity for cancer cells and the antitumor activity of pyrrolidine compound. The binding reaction between purpure-18-imide and bovine serum albumins (BSA) in aqueous was studied by fluorescence and UV-Vis absorption spectra. The research results indicated that the combination reaction of them was a single static quenching process. In aqueous, purpure-18-imide strongly bound BSA with molar ratio of 1:1. The binding constant K0 was 5.386 x 10(5) L x mol(-1). The shortest binding distance (r = 3.54 nm) and energy transfer efficiencies (E = 0.26) between donor (BSA) and acceptor (purpure-18-imide) were obtained by Forster's nonradiative energy transfer mechanism.

A new detection method for obtaining collisional transition probabilities P(E',E) of highly vibrationally excited molecules in the gas phase is presented. The technique employs energy-selective probing of the time-dependent vibrational population distribution by "kinetically controlled selective fluorescence (KCSF)". We present experimental results for a test system, the collisional deactivation of toluene by argon, where we use the well-known "kinetically controlled selective ionization (KCSI)" scheme as a reference for comparison. A newly designed setup is employed that allows simultaneous detection of fluorescence and ionization signals under identical experimental conditions ("kinetically controlled selective fluorescence and ionization = KCSFI"). For the system toluene + argon it is demonstrated that KCSF and KCSI yield identical results. A rate-equation model is presented to understand common features and differences of both approaches. The fluorescence detection scheme shows promise for future investigations on collisional energy transfer. The experimental setup is simpler, because it requires no additional ionization wavelength. This will hopefully give access to the P(E',E) of systems where, e.g., ionization schemes are difficult to implement due to short wavelengths required for the ionization step. A few examples will be outlined briefly.

Terbium (III) can react with Pefeoxacin (PEFX) to form 1:2 coordination compound in a near neutralmedium. Based on this reaction, we proposed a simple, rapid, and sensitive new method for the fluorimetric determination of Pefeoxacin and investigated the reaction conditions in detail; meanwhile we discussed the efficiency and mechanism of the intramolecular energy transfer. The results show that in the range of 0.01-1.67 microg x mL(-1), there is a linear relationship between the concentration of PEFX and its fluorescence intensity, and the regression equation is F = 120.01 C + 2.608, with the detection limit of 0.01 microg x mL(-1), the recovery within the range of 95.2%-101.7%, and the RSD of 2.9%-3.0%. This method has been successfully applied to the determination of Pefeoxacin in untreated human serum and gelatine capsule.