Correlation between fluorescence intermittency and spectral diffusion in single semiconductor quantum dots

We find a correlation between the dynamics of fluorescence intermittency and spectral diffusion in the spectroscopy of single CdSe nanocrystal quantum dots (QD). A statistical analysis of the data suggests two populations of blinking events: blinking followed by large spectral diffusion shifts and blinking with small or no spectral shifts. Although unexpected from earlier studies, the correlation between blinking and spectral shifting is consistent with a model of QD ionization as the mechanism for the blinking event, followed by a redistribution of local electric fields that results in spectral shifting.     

Enhanced fluorescence intermittency of CdSe-ZnS quantum-dot clusters

Fluorescence intermittency, or blinking, of individual close-packed clusters containing two or more CdSe-ZnS quantum dots (QDs) was investigated. The QD clusters exhibited rapid, intense blinking that was distinct from that of isolated QDs blinking independently. This enhanced blinking is suggested to occur when the QDs in the cluster become electronically coupled. The nature of this coupling is not known, though electrons trapped from QDs when they blink off may play a role by altering the electronic environment of neighboring QDs and enhancing their fluorescence properties.     

Fluorescence Intermittency in Monolayer WSe_2

Fluorescence intermittent dynamics of single quantum emitters in monolayer WSe2 are investigated via measuring spectrally resolved time traces and time-dependent fluorescence intensity trajectories. Analysis of fluorescence trajectories and spectral shifting reveal a correlation between the fluorescence intermittency and spectral diffusion.A model of an inverse power law can be used to understand the observed blinking dynamics.     

Manifestation of tunneling TLS dynamics of a polymer matrix in single-molecule fluorescence blinking

Blinking (stochastic intermittence) of fluorescence is a quite common phenomenon that accompanies the emission of single quantum objects-organic chromophore molecules, quantum dots, and nanocrystals. It is demonstrated that fluorescence blinking of single organic molecules embedded into a polymer matrix including the occurrence of “grey” states is due to tunneling transitions in the two-level systems (TLSs) of the matrix. The repeated registration of fluorescence excitation spectra of single molecules (SMs) is used for our analysis. The statistics of fluorescence blinking of an SM is directly related to conformational changes (tunneling transitions in TLSs) in its immediate vicinity. Individual parameters of the corresponding elementary excitation are also determined.     

界面电子转移对量子点荧光闪烁行为的影响

利用激光扫描共聚焦显微系统分别测量了CdSe/ZnS量子点在SiO2玻片表面、铟锡氧化物(ITO)纳米粒子表面和聚甲基丙烯酸甲酯(PMMA)薄膜表面上的荧光闪烁行为.研究发现,不同界面环境中量子点的亮态发光持续时间的概率密度都服从指数修正的幂律分布P(t)∝t-αexp(-t/μ).与处于SiO2玻片表面的情况相比,在ITO表面上的单量子点具有非常短暂的亮态发光持续时间,而在PMMA表面的单量子点亮态发光持续时间最长.这种荧光闪烁行为的不同主要归因于量子点与三种材料之间的界面电子转移特性.     

利用N型半导体纳米材料抑制单量子点的荧光闪烁特性

利用N型半导体纳米材料氧化铟锡(ITO)作为单CdSe/ZnS量子点的基质来抑制单量子点的荧光闪烁特性. 实验采用激光扫描共聚焦显微成像系统测量了单量子点荧光的亮、暗态持续时间的概率密度分布的指数截止的幂律特性, 并与直接吸附在SiO₂玻片上的单CdSe/ZnS量子点的荧光特性进行比较. 研究发现处于ITO中的单量子点比SiO₂玻片上的单量子点荧光亮态持续时间提高两个数量级, 掺杂于ITO中的单量子点的荧光寿命约减小为SiO₂玻片上的单量子点的荧光寿命的41%, 并且寿命分布宽度变小50%.     

Fluorescence intermittency in self-assembled InP quantum dots

Fluorescence intermittency in InP self-assembled dots is investigated by means of far field imaging and single dot spectroscopy. Based on our observation that blinking dots are found in the vicinity of scratches and the blinking frequency is drastically enhanced under a near-infrared laser irradiation, we attribute the origin of the fluorescence intermittency to a local electric field due to a carrier trapped at a deep localized center in the Ga0.5In0.5P matrix. The validity of this explanation is confirmed by a thermal activation-type behavior of the switching rate and artificial reproduction of the blinking phenomenon by an external electric field.     

Using the near-field coupling of a sharp tip to tune fluorescence-emission fluctuations during quantum-dot blinking

We demonstrate that the cycling between internal states of quantum dots during fluorescence blinking can be used to tune the near-field coupling with a sharp tip. In particular, the fluorescence emission from states with high quantum yield is quenched due to energy transfer, while that from low-yield states is elevated due to field enhancement. Thus, as a quantum dot blinks, its emission fluctuations are progressively suppressed upon approach of a tip.     

Fluorescence decay time of single semiconductor nanocrystals

We present fluorescence decay measurements of single ZnS covered CdSe nanocrystals. It is shown that the fluorescence decay time is fluctuating during the investigation leading to a multiexponential decay even for a single nanocrystal. In combination with measurements of the fluorescence blinking behavior we find that a high fluorescence intensity is correlated with a long fluorescence decay time. This is consistent with a model of fluctuating nonradiative decay channels leading to variable dynamic quenching processes of the excited state.     

From quantum light emitted by single molecule to classical light emitted by molecular ensemble

Fluorescence from an ensemble of noninteracting molecules excited by continuous laser radiation is considered as a glow of M individual molecules. The expression that governs the autocorrelation function (ACF) of fluorescence from M molecules is derived based on the theoretical expression that governs the ACF of fluorescence from a single-molecule. Fluctuations of blinking fluorescence and its statistical properties are analyzed using a computer simulation. The results of statistical analysis of the computer experiment are in good agreement with the derived formula that governs the ACF of fluorescence from several molecules.     

Sub-poissonian statistics of fluorescence photons of a single molecule

The method previously used by the author for the measurement and calculation of the distribution function w _N(T)of fluorescence photons from a single two-level atom that is continuously excited by laser light and has a unity fluorescence quantum yield is generalized to the case of a single molecule whose fluorescence quantum yield is smaller than unity and to the case of a three-level molecule whose fluorescence is blinking. The functions w _N(T) calculated for these two cases demonstrate a sub-Poissonian distribution of fluorescence photons.     

Surface-enhanced emission from single semiconductor nanocrystals

The fluorescence behavior of single CdSe(ZnS) core-shell nanocrystal (NC) quantum dots is dramatically affected by electromagnetic interactions with a rough metal film. Observed changes include a fivefold increase in the observed fluorescence intensity of single NCs, a striking reduction in their fluorescence blinking behavior, complete conversion of the emission polarization to linear, and single NC exciton lifetimes that are >10(3) times faster. The enhanced excited state decay process for NCs coupled to rough metal substrates effectively competes with the Auger relaxation process, allowing us to observe both charged and neutral exciton emission from these NC quantum dots.     

Blinking of single molecules in various environments

The fluorescence intermittency of various dye molecules in different environments was studied by wide-field fluorescence microscopy. The present work focuses on the analysis of long dark periods that are not due to triplet states. It is shown that the distributions of the length of dark periods follow power laws for all systems studied here. Furthermore, blinking kinetics is strongly influenced by the gas atmosphere to which the molecules are exposed. The presence of oxygen is of crucial importance.     

Direct measurement of the photon statistics of a triggered single photon source

We studied intensity fluctuations of a single photon source relying on the pulsed excitation of the fluorescence of a single molecule at room temperature. We directly measured the Mandel parameter Q(T) over 4 orders of magnitude of observation time scale T by recording every photocount. On time scale of a few excitation periods, sub-Poissonian statistics is clearly observed and the probablility of two-photons events is 10 times smaller than Poissonian pulses. On longer times, blinking in the fluorescence, due to the molecular triplet state, produces an excess of noise.     

Environment-dependent blinking of single semiconductor nanocrystals and statistical aging of ensembles

We compare decreasing fluorescence signals from ensembles with the blinking statistics of individual nanocrystals in various environments. For most substrates, the ensemble decay follows a power-law of time, the exponent being the difference of the power-law exponents of on- and off-time distributions. The decay exponent is also found to depend on substrate. We discuss possible mechanisms for this dependence, in conjunction with previously published models.     

功能性ZnS纳米荧光探针的合成及其光谱性质研究

报道了功能性ZnS纳米荧光探针的合成与表征 ,同时研究了纳米粒子的荧光衰变寿命 ,考察了该纳米荧光探针的吸收光谱和荧光发射光谱特性。与此同时 ,研究了不同蛋白质和核酸对该纳米荧光探针的吸收光谱和荧光发射光谱的影响 ,实验表明 ,蛋白质的加入使得该纳米荧光探针的吸收和发射光谱强度均增强 ,而核酸的加入则使其吸收和发射光谱强度均减弱。据此 ,可望将该纳米荧光探针应用于生物大分子的分析测定     

Statistics of photocounts of blinking fluorescence of quantum dots

The blinking of quantum dots under the action of laser radiation is described based on a model of a binary (two-state) renewal process with on (fluorescent) and off (non fluorescent) states. The T _on and T _off sojourn times in the on and off states are random and power-law distributed with exponents 0 < α < 1 and 0 < β < 1; the averages of the on and off times are infinite. As a consequence of this, the Gaussian statistics is inapplicable and the process is described using a more general statistics. An equation for the density of distribution p(t _on|t) of the total on time during the observation time t is derived that contains derivatives of fractional orders α and β. A solution to this equation is found in terms of fractional stable distributions. The Poisson transform of the density p(t _on|t) leads to the photon counting distribution and determines the fluorescence statistics. It is demonstrated that, if a blinking process with exponents α < β is implemented, then, at fairly long times, the on time will considerably prevail over the off time, i.e., blinking will be suppressed. This behavior is evidenced by the types of distributions of the total fluorescence time, the decay of relative fluctuations, and the Monte Carlo simulated trajectories of the process.     

Sub- and super-Poissonian photon statistics of single-molecule fluorescence blinking

An analysis of intermittent fluorescence is presented for a single molecule driven by a continuous-wave laser field. The interruptions of fluorescence are caused by transition of the molecule to a triplet state. A method previously developed to calculate photon distribution for continuous-wave fluorescence is applied to analyze photon statistics of fluorescence blinking. The probability w _N (T) of counting N photons over a time interval T is derived for intermittent fluorescence. The photons counted over relatively short intervals are found to have a sub-Poissonian (narrower than Poisson) distribution. The photon distribution over intervals longer than the mean off time has a complicated form with two maxima; i.e., a super-Poissonian (wider than Poisson) distribution is obtained.     

Application and New Developments in Fluorescence Spectroscopic Techniques in Studying Individual Molecules

Abstract

Techniques based on fluorescence have played a variety of roles in chemistry, physics, spectroscopy, medicine, nanotechnology, and biotechnology due to their high selectivity, sensitivity, simplicity, and fastness in spectroscopic and imaging measurements. While detecting fluorescence from individual molecules by fluorescence‐based techniques, poor signal, limited lifespan of fluorophores, trade‐off between time resolution, and the level of detail of information were few major concerns. Ultrasensitive detectors permit the combination of the high time resolution of single photon counting devices with the large field of view and spectral resolution allowed by two‐dimensional detectors. Photobleaching and on‐off blinking of fluorophores can be improved dramatically by chemical modifications or changing the reagents. New ways of controlling local fields such as optic, electric, magnetic, chemical, or biochemical environments take advantage of the noninvasiveness and high temporal and spatial resolution of single‐molecule fluorescence (SMF) to get a direct feedback of events at the nanometer scale in various domains of research. Some of the applications and new developments in fluorescence spectroscopic techniques in detecting, investigating, and/or manipulating individual molecules have been discussed.     

Single-Pair FRET Microscopy Reveals Mononucleosome Dynamics

We applied spFRET microscopy for direct observation of intranucleosomal DNA dynamics. Mononucleosomes, reconstituted with DNA containing a FRET pair at the dyad axis and exit of the nucleosome core particle, were immobilized through a 30 bp DNA tether on a polyethyleneglycol functionalized slide and visualized using Total Internal Reflection Fluorescence microscopy. FRET efficiency time-traces revealed two types of dynamics: acceptor blinking and intramolecular rearrangements. Both Cy5 and ATTO647N acceptor dyes showed severe blinking in a deoxygenated buffer in the presence of 2% βME. Replacing the triplet quencher βME with 1 mM Trolox eliminated most blinking effects. After suppression of blinking three subpopulations were observed: 90% appeared as dissociated complexes; the remaining 10% featured an average FRET efficiency in agreement with intact nucleosomes. In 97% of these intact nucleosomes no significant changes in FRET efficiency were observed in the experimentally accessible time window ranging from 10 ms to 10’s of seconds. However, 3% of the intact nucleosomes showed intervals with reduced FRET efficiency, clearly distinct from blinking, with a lifetime of 120 ms. These fluctuations can unambiguously be attributed to DNA breathing. Our findings illustrate not only the merits but also typical caveats encountered in single-molecule FRET studies on complex biological systems.