CdTe量子点与罗丹明B水溶液体系下的双光子激发荧光共振能量转移

采用时间分辨荧光光谱技术研究了在双光子激发下不同尺寸的量子点与罗丹明B 之间的荧光共振能量转移. 研究结果表明, 在800 nm的双光子激发条件下, 体系间能量转移效率随着供体吸收光谱与受体荧光光谱的光谱重叠程度增加而增加; 理论分析表明, 供体和受体间的Förster半径增加是导致其双光子能量转移效率增大的物理原因. 同时, 研究了罗丹明B浓度对荧光共振能量转移效率的影响. 研究结果表明, 量子点的荧光寿命随着罗丹明B浓度的增加而减小; 量子点与罗丹明B之间的荧光共振能量转移效率随着罗丹明B浓度的增加而增加; 当罗丹明B浓度为3.0×10^-5 mol·L^-1时, 双光子荧光共振能量转移效率为40.1%.     

Energy transfer probe for the characterization of luminescent photonic crystals morphology

We evaluated energy transfer probe for the characterization of luminescent photonic crystals morphology. It demonstrates a monomolecular filming of fluorescent chelated complexes inside photonic crystal voids. Either direct fluorescence quenching of excited fluorescent donors by the acceptors in case of low concentration of fluorescent centers or fluorescence quenching accelerated by energy migration over fluorescent donor ions in case of 100% of sites occupied by donors is demonstrated in 2D space contrarily to the powder samples of the same complexes where energy transfer occurs in 3D space.     

Nonradiative and radiative Förster energy transfer between quantum dots

We theoretically study nonradiative and radiative energy transfer between two localized quantum emitters, a donor (initially excited) and an acceptor (receiving the excitation). The rates of nonradiative and radiative processes are calculated depending on the spatial and spectral separation between the donor and acceptor states and for different donor and acceptor lifetimes for typical parameters of semiconductor quantum dots. We find that the donor lifetime can be significantly modified only due to the nonradiative Förster energy transfer process at donor–acceptor separations of approximately 10 nm (depending on the acceptor radiative lifetime) and for the energy detuning not larger than 1–2 meV. The efficiency of the nonradiative Förster energy transfer process under these conditions is close to unity and decreases rapidly with an increase in the donor–acceptor distance or energy detuning. At large donor–acceptor separations greater than 40 nm, the radiative corrections to the donor lifetime are comparable with nonradiative ones but are relatively weak.     

Radiationless intermolecular energy transfer with participation of acceptor excited triplet states

Radiationless energy transfer between like and unlike molecules has been experimentally studied under conditions where acceptor molecules have been excited to the triplet state Homogeneous singlet-triplet-triplet migration has been discovered in highlyconcentrated chlorophyll “a” and pheophytin “a” solutions in castor oil at 183 K by measuring the variation of pigment relative quantum yields of fluorescence and triplet state formation as a function of exciting pulse intensity. Heterogeneous single-triplet-triplet energy transfer has been observed in solid solutions of different complex organic molecules (perylene + phenanthrene, Na-fluorescein+chlorophyll “a”, pyrene+Mg-phthalocyanine) as the fluorescent donor state quenching in the presence of acceptor triplet-excited molecules. Primary emphasis is placed on a direct observation of the effect of energy transfer on the excited-state lifetime of the donor. The benzophenone phosphorescence quenching (shortening of phosphorescence lifetime) in the presence of Mg-mesoporphyrin triplet molecules has been found to be caused by the heterogeneous triplet-triplet-triplet energy transfer. Good agreement of the theoretical and experimental results permits us to conclude that all types of observed transfer processes are described by the Förster-Galanin theory for dipole-dipole radiationless energy transfer with no additional assumptions.     

Two-Photon Excited Fluorescence Energy Transfer: A Study Based on Oligonucleotide Rulers

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 [1]. 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.     

“Discrete shell model” for analysing time-resolved energy transfer in solids

We present a new mathematical method to describe the time-resolved fluorescence response of donors and acceptors for the case of energy transfer in crystals, taking into account energy diffusion within the donor system, direct donor-acceptor transfer with any arbitrary dependence of the transfer probability on distance, and the discrete crystal structure. For the first time we give the fluorescence responses for elevated temperatures including transfer of optical excitation energy from the acceptors back to the donors. Furthermore, the competition between neighbouring acceptors (excitation sinks), important for most real acceptor concentrations, is taken into account. The fluorescence responses of donors and acceptors to four commonly realized excitation conditions are given explicitly to any desired degree of precision as sums of exponential functions; they are based on the numerical solution of an eigenproblem. The method is designed to be used in computer analysis of time-resolved fluorescence data.     

Dynamics of fluorescence of solid solutions of quantum dots and the organic dye DODCI: Experiment and numerical simulation

The fluorescence of solid solutions of CdSe/ZnS quantum dots and the organic dye DODCI is investigated. It is shown that nonradiative transfer of electronic excitation energy to dye molecules, which with some probability lose their acceptor properties as a result of photoisomerization or photodegradation, is responsible for a significant increase in the fluorescence intensity of a donor. The degree of polarization of the donor fluorescence attains values exceeding 0.5, which is due to the difference in the fluorescence quantum yields of donors with different orientations of the oscillator with respect to the electric vector of an excitation light wave. A numerical simulation of the experimentally observed dependences is performed.     

Förster resonant energy transfer in quantum dot layers

Förster resonant energy transfer (FRET) in quantum dot (QD) layer structures has been analyzed. Small and large colloidal CdTe QDs were used as donors and acceptors, respectively. A FRET theory for random donor/acceptor distributions in two dimensions, taking into account exclusion zones around the donors, was applied to characterize FRET in a mixed monolayer. The exclusion zones provide a possibility to include the QD size in the FRET analysis and to determine its impact on the FRET efficiency. The acceptor concentration dependence of the FRET efficiency can also be described within this theory. In a separate donor/acceptor layer structure the distance dependence of the FRET efficiency as well as the acceptor enhancement was investigated. Both were found to agree well with the model of FRET between donor and acceptor layers.     

Investigation of a Fluorescence Signal Amplification Mechanism Used for the Direct Molecular Detection of Nucleic Acids

A fluorescence signal amplification mechanism allowing detection limits for DNA in the zeptomolar range was investigated. Photophysical properties of the molecular system were studied in order to better explain the signal amplification that is observed. We show that the confinement of a fluorescent DNA hybridization transducer in aggregates improves its quantum yield and photostability. Furthermore, we show that the combination of the resonance energy transfer occurring within the aggregates with the use of a conjugated polymer as the hybridization transducer and donor allows ultrafast and efficient energy coupling to the aggregates and can lead to the excitation of a large number of acceptors by only one donor.     

Energy transfer from mesitylene and benzene to 9,10-diphenylanthracene. The influence of donor concentration

Nonradiative energy transfer from neat and diluted mesitylene and benzene to 9,10-diphenylanthracene (DPA) was studied in high concentrations of the donors (mesitylene in the range of 7.2 to 0.72 M and benzene from 11.2 to 1.12 M).A new set of equations is presented for steady-state analysis of the emission of the excited monomer and excimer in the presence or absence of an acceptor.Experimental values obtained for the ratio of the fluorescence emission quantum yield of the donor in the absence and presence of the acceptor (φ₀/φ)_X as a function of the acceptor concentration C_DPA show positive deviations from linearity that are compatible with the use of an effective radius R_eff greater than the collisional radius R_e, suggesting the existence of energy migration in the donor competing with the diffusion process.     

Binding and Fluorescence Resonance Energy Transfer (FRET) of Ruthenium(II)-Bipyridine-Calixarene System with Proteins—Experimental and Docking Studies

The investigation of the interaction of ruthenium(II)-bipyridine-tert-butylcalix[4]arene complexes (Rubc2 and Rubc3) with proteins (BSA and ovalbumin) using absorption, emission, excited state lifetime and circular dichroism techniques and by docking studies show that luminophore-receptor system bind strongly with proteins. An enhancement of absorption as well as emission intensity of Ru(II)-calixarene complexes in the presence of proteins, but the quenching of the emission intensity of proteins in the presence of Ru(II)-calixarene complexes are the interesting observations. The enhancement of emission intensity of Ru(II)-calixarene complex, in the presence of proteins, is due to the fluorescence resonance energy transfer (FRET) from protein to Ru(II)-calixarene complex. Among the two Ru(II)-calixarene complexes synthesized Rubc3 has more efficient binding and energy transfer than Rubc2 and BSA, with a large cavity size, has the advantage for binding over ovalbumin. Docking studies reveal that the presence of tert-butylcalix[4]arene moiety in Ru(II)-calixarene complexes facilitates binding with proteins. After the binding of Rubc2 and Rubc3 with proteins, the nearby fluorophores present in proteins are in optimal distance from the ruthenium centre for efficient FRET process to occur.     

金属卟啉分子组装体系中的三线态能量传递

利用稳态和瞬态光谱技术研究了人工组装锌卟啉(ZnP)-苯桥(BB)-铁卟啉(Fe(Cl)P)超分子体系中给体三线态到受体的能量传递及其机理。结果表明体系中存在着由给体ZnP三线态向受体Fe(Cl)P的超快能量传递过程,在室温和低温下通过激发给体ZnP,其单线态的激发能经由系间窜越过程使其三线态布居,在受体Fe(Cl)P存在的情况下,位于给体三线态的激发能经由桥联分子B传递到受体Fe(Cl)P,室温下传递速率为7.2×105s-1。由于体系中给体到受体之间的空间距离约为2.5nm,由给体-受体直接耦合引起的传递机理可以排除,由桥联分子媒介的超交换机理是该能量传递过程的主要物理机理。     

Tuning energy transfer efficiency in quantum dots mixture by controling donor/acceptor ratio

Improving the emission performance of colloidal quantum dots (QDs) is of paramount importance for their applications on light-emitting diodes (LEDs), displays and lasers. A highly promising approach is to tune the carrier recombination channels and lifetime by exploiting the energy transfer process. However, to achieve this precise emission optimization, quantitative modulation on energy transfer efficiency is highly desirable but still challenging. Here, we demonstrate a convenient approach to realize tunable energy transfer efficiency by forming QDs mixture with controllable donor/acceptor (D/A) ratio. With the mixing ratio ranging from 16/1 to 1/16, the energy transfer efficiency could be effectively tuned from near zero to ~70%. For the high mixing ratio of 16/1, acceptors obtain adequate energy supplied by closely surrounding donors, leading to~2.4-fold PL enhancement. While for the low mixing ratio, the ultrafast and efficient energy extraction process directly suppresses the multi-exciton and Auger recombination in the donor, bringing about a higher threshold. The facile modulation of emission performance by controllably designed mixing ratio and quantitatively tunable energy transfer efficiency will facilitate QD-based optoelectronic and photovoltaic applications.     

芘四磺酸四钠盐光物理性质的研究

本文利用吸收光谱、荧光光谱以及时间分辨瞬态吸收光谱研究芘四磺酸四钠盐的光物理特性.研究结果表明,芘四磺酸四钠盐在水溶液中的荧光量子产额高达0.54.在纳秒时间分辨瞬态吸收光谱研究中发现芘四磺酸四钠盐具有很长的三重态寿命(3.5ms),这有利于将其作为光敏化分子把激发能传给其它受体分子;芘四磺酸四钠盐被光激发后,可能发生双光子吸收过程并产生芘四磺酸四钠盐阳离子,其吸收峰为460nm和505nm.由实验结果,我们给出了其动力学过程的解释.     

Fluorescence resonance energy transfer: FRET studies of ligand binding to cell surface receptors

We describe a simple optical system employing fluorescence resonance energy transfer (FRET) to identify potential binding domains on the macrophage scavenger receptor for the ligand maleylated bovine serum albumin (mal-BSA). Using a plasma membrane vesicle system, we placed donor probes on the ligand and acceptor probes in the membrane to determine the distance of bound ligand from the cell surface. Two donors and three acceptors were employed. Transfer between ligand covalently modified with multiple dansyl molecules and hexadecanoylaminoeosin in the membrane yielded a distance of 46.5 ± 7.5 å; transfer from the same type of donors to octadecylrhodamine B in the membrane gave a distance of 58.5 ± 3.0 å. No transfer was observed between ligand mono-labeled with fluorescein and l,l′-dioctadecyl-3,3,3′,3′-tetramethylindocarbocyanineperchlorate in the membrane. This suggests that the orientation of mal-BSA bound to the receptor places the fluorescein probe too far from the lipid surface to experience energy transfer. The distance information identifies a potential location for the binding site, which can be compared to structural information about the receptor and used to extract a binding sequence.     

Assignment of the low frequency vibrations in electron donor-acceptor complexes by ultrafast coherent spectroscopy

Coherent oscillations in a spontaneous fluorescence of electron donor-acceptor (EDA) complexes in solution were observed with three different acceptors: tetracyanoethylene (TCNE), chloranil, and fluoranil with various electron donors. Different oscillatory frequencies were observed for the complexes with different acceptors. The similar frequencies are observed for the complexes with different donors and a common acceptor: 155 cm⁻¹ (period of 215 fs) for the complexes with TCNE, 178 cm⁻¹ (187 fs) for the complexes with chloranil, and 212 cm⁻¹ (157 fs) for the complex with fluoranil. It is concluded, that the out-of-plane vibrational mode of the acceptor, having b_3u symmetry, is responsible for the observed oscillations in all three cases (three acceptors). The time-dependent fluorescence spectrum for the TCNE - HMB complex is reconstructed and the fluorescence peak-shift correlation function is obtained. The ultrafast relaxation of the fluorescence peak position is superimposed with the oscillatory component. The main component of the spectral relaxation of 115 fs is attributed mainly to the intramolecular vibrational energy redistribution process in both donor and acceptor parts of the complex. The oscillatory behavior of the fluorescence peak position demonstrates the modulation of the transition frequency during the vibrations. The modulation of the mean transition moment is observed as well. Thus two mechanisms are observed to be responsible for the oscillations: the modulation of the transition frequency and the modulation of the mean transition moment by the vibrational motion.     

Mechanisms of Fluorescence Quenching in Donor—Acceptor Labeled Antibody—Antigen Conjugates

The cyanine dyes Cy5 and Cy5.5 are presented as a new long wavelength-excitable donor-acceptor dye pair for homogeneous fluoroimmunoassays. The deactivation pathways responsible for the quenching of the fluorescence of the antibody-bound donor are elucidated. Upon binding of the donor dye to the antibodies at low dye/protein ratios, its fluorescence quantum yield rises to unity. Higher dye/protein ratios lead to progressive aggregation of the dyes, which results in quenching of monomer fluorescence due to resonance energy transfer (RET) from the monomers to the nonfluorescent dimers. The dependence of the quenching efficiency on the labeling ratio is described quantitatively by assuming a Poisson distribution of the dyes over the antibodies. The maximum fluorescence intensity per antibody is obtained at a labeling ratio of 4. Upon formation of the antibody-antigen complex, electron transfer and RET to the antigen-bound acceptor dye occur. Steady-state and time-resolved fluorescence measurements reveal that approximately 50% of the donor quenching is due to RET, while the residual quenching effect is caused by the static quenching process.     

Time-gated microscopic energy transfer measurements for probing mitochondrial metabolism

Spectroscopic and microscopic methods for probing mitochondrial malfunction were established using cultivated endothelial cells from the calf aorta and various inhibitors of the respiratory chain, which is located at the inner mitochondrial membrane. Time-gated fluorescence spectroscopy was used to measure autofluorescence of the coenzyme NADH as well as “energy transfer efficacy” from excited NADH molecules (energy donor) to the mitochondrial marker rhodamine-123 (energy acceptor). Autofluorescence usually exhibited a weak increase after specific inhibition of enzyme complexes of the respiratory chain. In contrast, a pronounced increase in energy transfer efficacy was observed after inhibition of the same enzyme complexes. The detection of donor (NADH) and acceptor (R123) fluorescence in different nanosecond time gates following the exciting laser pulses enhances selectivity and improves quantification of energy transfer measurements. Therefore, timegated energy transfer spectroscopy is suggested to be an appropriate tool for probing mitochondrial malfunction.     

Interaction among Cadmium Sulfide Nanoparticles, Acridine Orange, and Deoxyribonucleic Acid in Fluorescence Spectra and a Method for Deoxyribonucleic Acid Determination

Some studies on quantum dots (QD) as donors that enhance the fluorescence of a dye as an acceptor through fluorescence resonance energy transfer (FRET) have been reported. However, in the present work we discovered that CdS quantum dots sharply quenched the fluorescence of acridine orange (AO). Also, DNA enhanced the fluorescent signals of AO quenched by CdS. The extents of enhancement were in good proportion to the DNA concentrations. Based on this, a sensitive method was employed to determine DNA with both good selectivity and sensitivity. The calibration curve was linear over 60-4,000 ng mL(-1) and the determination limit (3sigma) was 4.39 ng mL(-1).