Fluorescence Intermittency Research Articles

Fluorescence super-resolution microscopy via fluctuation-based multi-route synergy.

Fluorescence fluctuation-based super-resolution microscopy (FF-SRM) is an economical and widely applicable technique that significantly enhances the spatial resolution of fluorescence imaging by capitalizing on fluorescence intermittency. However, each variant of FF-SRM imaging has inherent limitations. This study proposes a super-resolution reconstruction strategy (synSRM) by synergizing multiple variants of the FF-SRM approach to address the limitations and achieve high-quality and high-resolution imaging. The simulation and experimental results demonstrate that, compared to images reconstructed using single FF-SRM algorithms, by selecting suitable synSRM routes according to various imaging conditions, further improvements of the spatial resolution and image reconstruction quality can be obtained for super-resolution fluorescence imaging.

Generalized Einstein relation for aging processes

Physical aging appears in many systems ranging from glassy/granular materials, blinking quantum dots to laser-cooled atoms. Aging is a process with three fingerprints: (i) slow, non-exponential relaxation, (ii) breaking of time-translation-invariance, and (iii) dynamical scaling. Here, we show that all these features are present in our minimal Langevin model for aging. A natural extension of the Einstein relation, which was expected to be true in an equilibrium state, is conjectured to hold in aging processes where both the damping and the temperature decrease with time in power-law forms. The generalized Einstein relation can be used to tackle the difficult problem of determining non-ergodic behaviours. The model shows a power-law-type diffusion away from the critical point and a logarithmic Sinai-type ultra-slow diffusion at the critical point. Application to granular gases is also discussed.

Weakly Confined Organic-Inorganic Halide Perovskite Quantum Dots as High-Purity Room-Temperature Single Photon Sources.

Colloidal perovskite quantum dots (PQDs) have emerged as highly promising single photon emitters for quantum information applications. Presently, most strategies have focused on leveraging quantum confinement to increase the nonradiative Auger recombination (AR) rate to enhance single-photon (SP) purity in all-inorganic CsPbBr3 QDs. However, this also increases the fluorescence intermittency. Achieving high SP purity and blinking mitigation simultaneously remains a significant challenge. Here, we transcend this limitation with room-temperature synthesized weakly confined hybrid organic-inorganic perovskite (HOIP) QDs. Superior single photon purity with a low g(2)(0) < 0.07 ± 0.03 and a nearly blinking-free behavior (ON-state fraction >95%) in 11 nm FAPbBr3 QDs are achieved at room temperature, attributed to their long exciton lifetimes (τX) and short biexciton lifetimes (τXX). The significance of the organic A-cation is further validated using the mixed-cation FAxCs1-xPbBr3. Theoretical calculations utilizing a combination of the Bethe-Salpeter (BSE) and k·p approaches point toward the modulation of the dielectric constants by the organic cations. Importantly, our findings provide valuable insights into an additional lever for engineering facile-synthesized room-temperature PQD single photon sources.

Quantum dots: modern methods of synthesis and optical properties

Quantum dots are the most exciting representatives of nanomaterials. They are synthesized using advanced methods of nanotechnology pertaining to both inorganic and organic chemistry. Quantum dots possess unique physical and chemical properties; therefore, they are used in very different fields of physics, chemistry, biology, engineering and medicine. It is not surprising that the Nobel Prize in chemistry in 2023 was given for discovery and synthesis of quantum dots. This review addresses modern methods for the synthesis of quantum dots and their optical properties and practical applications. In the beginning, a short insight into the history of quantum dots is given. Many gifted scientists, including chemists and physicists, were engaged in these studies. The synthesis of quantum dots in solid and liquid matrices is described in detail. Quantum dots are well-known owing to their unique optical properties; that is why the attention in the review is focused on the quantum-size effect. The causes for fascinating blinking of quantum dots and techniques for observation of a single quantum dot are considered. The last part of the review describes mportant applications of quantum dots in biology, medicine and quantum technologies.&lt;br&gt; The bibliography includes 772 references.

Efficient characterization of blinking quantum emitters from scarce data sets via machine learning

Single photon emitters are core building blocks of quantum technologies, with established and emerging applications ranging from quantum computing and communication to metrology and sensing. Regardless of their nature, quantum emitters universally display fluorescence intermittency or photoblinking: interaction with the environment can cause the emitters to undergo quantum jumps between on and off states that correlate with higher and lower photoemission events, respectively. Understanding and quantifying the mechanism and dynamics of photoblinking is important for both fundamental and practical reasons. However, the analysis of blinking time traces is often afflicted by data scarcity. Blinking emitters can photo-bleach and cease to fluoresce over time scales that are too short for their photodynamics to be captured by traditional statistical methods. Here, we demonstrate two approaches based on machine learning that directly address this problem. We present a multi-feature regression algorithm and a genetic algorithm that allow for the extraction of blinking on/off switching rates with ⩾85% accuracy, and with ⩾10× less data and ⩾20× higher precision than traditional methods based on statistical inference. Our algorithms effectively extend the range of surveyable blinking systems and trapping dynamics to those that would otherwise be considered too short-lived to be investigated. They are therefore a powerful tool to help gain a better understanding of the physical mechanism of photoblinking, with practical benefits for applications based on quantum emitters that rely on either mitigating or harnessing the phenomenon.

Multiphoton emission of single CdZnSe/ZnS quantum dots coupled with plasmonic Au nanoparticles.

The fluorescence blinking and low multiphoton emission of quantum dots (QDs) have limited their application in lasing, light-emitting diodes, and so on. Coupling of single QDs to plasmonic nanostructures is an effective approach to control the photon properties. Here plasmon-exciton systems including Au nanoparticles and CdZnSe/ZnS QDs were investigated at the single particle level. With the modulation of the local electromagnetic field, the fluorescence intensity of single QDs is increased, accompanied by a significant suppression in blinking behavior, and the lifetime is shortened from 15 ns to 2 ns. Moreover, the second-order photon intensity correlation at zero lag time g2(0) of coupled single QDs is larger than 0.5, indicating an increased probability of multiphoton emission. The enhancement factors of radiative and nonradiative decay rates of QDs coupled with Au nanoparticles are calculated. The sharply increased radiative decay rate can be comparable to the nonradiative Auger rate, leading to dominated multiple exciton radiative recombination with PL intensity enhancement, suppressed blinking, lifetime shortening, and multiphoton emission. The results of the exciton decay dynamics and emission properties of single QDs in this work are helpful in exploring the mechanism of plasmon-exciton interaction and optoelectronic application of single QDs.

Evidence of carrier diffusion between emission states in CdSe/ZnS core-shell quantum dots: a comprehensive investigation combining fluorescence lifetime correlation spectroscopy (FLCS) and single dot photoluminescence studies.

Investigation of carrier dynamics in CdSe/ZnS core-shell quantum dots (QDs) is performed using fluorescence-lifetime-correlation-spectroscopy (FLCS) and single-dot PL blinking studies. The origin of an emitted photon from a QD in an FLCS study is assigned to either an exciton state or trap state based on its excited state lifetime (τfl). Subsequently, two intrastate autocorrelation functions (ACFs) representing the exciton and trap states and one cross-correlation function (CCF) coupling these two states are constructed. Interestingly, the timescales of carrier diffusion (τR) show striking similarities across all three correlation functions, which further correlate with τR of the conventional FCS. However, ACFs notably deviate from the CCF in their μs progression patterns, with the latter showing growth, whereas the former ones display decay. This implies inter-state carrier diffusions leading to the QD blinking. Further study of single particle PL blinking on a surface-immobilized QD indicates shallow trap states near the band edge cause the blinking at low excitation power, while trion recombination becomes an additional contributing factor at higher pump power. Overall, the results highlight not only an excellent correlation between these two techniques but also the potential of our approach for achieving an accurate and comprehensive understanding of carrier dynamics in CdSe/ZnS QDs.

Room-Temperature Single-Photon Sources Based on Colloidal Quantum Dots: A Review.

Single-photon sources (SPSs) play a crucial role in quantum photonics, and colloidal quantum dots (CQDs) have emerged as promising and cost-effective candidates for such applications due to their high-purity single-photon emission at room temperature. This review focuses on various aspects of CQDs as SPSs. Firstly, a brief overview of the fundamental optical properties of CQDs is provided, including emission wavelength engineering and fluorescence intermittency, and their single-photon emission properties. Subsequently, this review delves into research concerning CQDs as SPSs, covering topics such as the coupling of single CQDs to microcavities, both in weak and strong coupling regimes. Additionally, methods for localizing and positioning CQDs are explored, which are critical for on-chip SPSs devices.

Biexciton Blinking in CdSe-Based Quantum Dots.

Experiments on single colloidal quantum dots (QDs) have revealed temporal fluctuations in the emission efficiency of the single-exciton state. These fluctuations, often termed "blinking", are caused by opening/closing of charge-carrier traps and/or charging/discharging of the QD. In the regime of strong optical excitation, multiexciton states are formed. The emission efficiencies of multiexcitons are lower because of Auger processes, but a quantitative characterization is challenging. Here, we quantify fluctuations of the biexciton efficiency for single CdSe/CdS/ZnS core-shell QDs. We find that the biexciton efficiency "blinks" significantly. The additional electron due to charging of a QD accelerates Auger recombination by a factor of 2 compared to the neutral biexciton, while opening/closing of a charge-carrier trap leads to an increase of the nonradiative recombination rate by a factor of 4. To understand the fast rate of trap-assisted recombination, we propose a revised model for trap-assisted recombination based on reversible trapping. Finally, we discuss the implications of biexciton blinking for lasing applications.

Fluorescence Intermittency of Quantum Dot–Organic Dye Conjugates: Implications for Alternative Energy and Biological Imaging

Quantum dot (QD)-organic dye couple chromophores are topical due to their applications in biology, catalysis, and energy. The maximization of energy transfer efficiency can be guided by the underlying Förster or Dexter mechanisms; however, the impact of fluorescence intermittency must also be considered. Here we demonstrate that the average ⟨ton⟩ and ⟨toff⟩ times of dye acceptors in coupled QD-dye chromophores are substantially affected by the donors' blinking behavior. With regard to biological imaging, this effect beneficially minimizes the photobleaching of the acceptor dye. The implications for alternative energy are less encouraging as the acceptors' capacity to store energy, using ⟨ton⟩/⟨toff⟩ as a metric, was reduced by as much as ∼95%. These detrimental effects can be mitigated by suppressing QD blinking via surface treatment. This study also demonstrates several instances of the nonconformity of QD blinking dynamics to a power law distribution, as a robust examination of the off times reveals log-normal behavior that is consistent with the Albery model.

Photonic Crystal Enhanced Fluorescence: A Review on Design Strategies and Applications.

Nanoscale fluorescence emitters are efficient for measuring biomolecular interactions, but their utility for applications requiring single-unit observations is constrained by the need for large numerical aperture objectives, fluorescence intermittency, and poor photon collection efficiency resulting from omnidirectional emission. Photonic crystal (PC) structures hold promise to address the aforementioned challenges in fluorescence enhancement. In this review, we provide a broad overview of PCs by explaining their structures, design strategies, fabrication techniques, and sensing principles. Furthermore, we discuss recent applications of PC-enhanced fluorescence-based biosensors incorporated with emerging technologies, including nucleic acids sensing, protein detection, and steroid monitoring. Finally, we discuss current challenges associated with PC-enhanced fluorescence and provide an outlook for fluorescence enhancement with photonic-plasmonics coupling and their promise for point-of-care biosensing as well monitoring analytes of biological and environmental relevance. The review presents the transdisciplinary applications of PCs in the broad arena of fluorescence spectroscopy with broad applications in photo-plasmonics, life science research, materials chemistry, cancer diagnostics, and internet of things.

FRET-Mediated Collective Blinking of Self-Assembled Stacks of CdSe Semiconducting Nanoplatelets

Förster resonant energy transfer (FRET) can be an efficient energy transfer mechanism between densely packed fluorescent emitters. It plays a key role in photosynthesis but may also be detrimental. In optoelectronic devices for instance, FRET funnels energy to quenching sites and favors losses. Here, we image individual self-assembled chains of stacked CdSe nanoplatelets and demonstrate fluorescence intermittency (blinking) of chain portions corresponding to a few tens of platelets. This collective blinking is attributed to the fluctuations of a quencher site, to which excitons are transferred by FRET migration from the surrounding platelets. We develop an analytical random walk model of the chain and show that an ensemble of platelets can be quenched collectively by a single site provided that its quenching (nonradiative recombination) rate is faster than the geometric mean of the radiative recombination rate and the transfer rate, which for self-assembled platelets would be of the order of (100 ps)−1.

Correlating Fluorescence Intermittency and Second‐Harmonic Generation in Single‐Colloidal Semiconductor Nanoplatelets

While most single‐nanocrystal spectroscopy experiments rely on fluorescent emission, recent years have seen an increasing number of experiments based on absorption and scattering, enabling to correlate those with fluorescence intermittency. Herein, it is shown that nonlinear scattering by second‐harmonic generation can also be measured from single CdSe/CdS core/shell nanoplatelets (NPLs) alongside fluorescence despite the weak scattering signal. It is shown that even under resonant two‐photon conditions the second‐harmonic scattering signal is uncorrelated with fluorescence intermittency and follows Poisson statistics.

Single-Molecule Peptide Identification Using Fluorescence Blinking Fingerprints.

The ability to identify peptides with single-molecule sensitivity would lead to next-generation proteomics methods for basic research and clinical applications. Existing single-molecule peptide sequencing methods can read some amino acid sequences, but they are limited in their ability to distinguish between similar amino acids or post-translational modifications. Here, we demonstrate that the fluorescence intermittency of a peptide labeled with a spontaneously blinking fluorophore contains information about the structure of the peptide. Using a deep learning algorithm, this single-molecule blinking pattern can be used to identify the peptide. This method can distinguish between peptides with different sequences, peptides with the same sequence but different phosphorylation patterns, and even peptides that differ only by the presence of epimerized residues. This study builds the foundation for a targeted proteomics method with single-molecule sensitivity.

A quantitative model of multi-scale single quantum dot blinking

We present a fundamentally new model of colloidal semiconductor quantum dot blinking. The blinking is caused by fluctuations of the non-radiative exciton relaxation rate, induced by variations of the electron–phonon coupling value.

Suppression of blinking in single CsPbBr3 perovskite nanocrystals through surface ligand exchange.

Photoluminescence blinking in individual semiconducting and perovskite quantum dots reflects reduced emission quantum yield and represents an obstacle towards quantum dot applications. One of the origins of blinking is the presence of surface structural defects that can function as charge traps. To reduce the defects the surface can be modified by, e.g., covering with ligands that are more strongly bound to the surface. Here, we report exchange of ligands on the CsPbBr3 perovskite nanocrystal surface and the effect of the exchange on photoluminescence blinking. Replacement of the oleic acid and oleylamine ligands which are used in the synthesis process with quaternary amine ligands leads to substantial increase of photoluminescence quantum yield. On single particle level this is reflected by significantly improved blinking characteristics. Statistical analysis using the probability density function shows that the ligand exchange leads to longer duration of ON-times and shorter OFF-times, as well as to the presence of a higher fraction of ON-time intervals. These characteristics are not affected by sample aging within three weeks. On the contrary, storage of the samples in solution for one-to-two weeks leads to further improvement of the ON-time interval fraction statistics.

Protein Conjugation Helped CdTe Quantum Dots for the Specific Labeling and Super‐Resolution Imaging of Lysosomes

AbstractSemiconductor quantum dots possess many attractive features such as high quantum yield, and narrow emission, and offer great advantages over organic dyes. However, the fluorescence intermittency and the cytotoxicity limit their biological applications. In this communication, we show a substantial increase in cell viability and the tuning of intermittency of water‐soluble CdTe quantum dots upon conjugation with bovine serum albumin (BSA), which helped them for their use as fluorescent probe for super resolution bioimaging of lysosomes. The quantum dots were not only found to be highly specific to stain the lysosome but also provided the super‐resolution image's resolution down to ∼60 nm, which is close to the smallest size of the lysosome. Photons per cycle were found to play a significant role in improving localization precision and thus improved the quality of the super resolution images.

Photonic crystal enhanced fluorescence emission and blinking suppression for single quantum dot digital resolution biosensing

While nanoscale quantum emitters are effective tags for measuring biomolecular interactions, their utilities for applications that demand single-unit observations are limited by the requirements for large numerical aperture (NA) objectives, fluorescence intermittency, and poor photon collection efficiency resulted from omnidirectional emission. Here, we report a nearly 3000-fold signal enhancement achieved through multiplicative effects of enhanced excitation, highly directional extraction, quantum efficiency improvement, and blinking suppression through a photonic crystal (PC) surface. The approach achieves single quantum dot (QD) sensitivity with high signal-to-noise ratio, even when using a low-NA lens and an inexpensive optical setup. The blinking suppression capability of the PC improves the QDs on-time from 15% to 85% ameliorating signal intermittency. We developed an assay for cancer-associated miRNA biomarkers with single-molecule resolution, single-base mutation selectivity, and 10-attomolar detection limit. Additionally, we observed differential surface motion trajectories of QDs when their surface attachment stringency is altered by changing a single base in a cancer-specific miRNA sequence.

Memory in quantum dot blinking.

The photoluminescence intermittency (blinking) of quantum dots is interesting because it is an easily measured quantum process whose transition statistics cannot be explained by Fermi's golden rule. Commonly, the transition statistics are power-law distributed, implying that quantum dots possess at least trivial memories. By investigating the temporal correlations in the blinking data, we demonstrate with high statistical confidence that there is nontrivial memory between the on and off brightness duration data of blinking quantum dots. We define nontrivial memory to be statistical complexity greater than one. We show that this memory cannot be discovered using the transition distribution. We show by simulation that this memory does not arise from standard data manipulations. Finally, we conclude that at least three physical mechanisms can explain the measured nontrivial memory: (1) storage of state information in the chemical structure of a quantum dot; (2) the existence of more than two intensity levels in a quantum dot; and (3) the overlap in the intensity distributions of the quantum dot states, which arises from fundamental photon statistics.

Shell-dependent blinking and ultrafast carrier dynamics in CdxZn1-xSeyS1-y@ZnS core/shell quantum dots

Fluorescent semiconductor quantum dots (QDs) are promising nanocrystals with excellent optical properties for applications in many fields. However, the severe blinking of QDs limits their practical applications that rely on single-dot emission. In this study, the shell-dependent blinking behavior and ultrafast carrier dynamics in CdxZn1-xSeyS1-y@ZnS core/shell QDs with smooth interface were investigated by using single particle fluorescence imaging and time-resolved spectroscopy. Our observations indicate that the on-time fraction of emission blinking for CdxZn1-xSeyS1-y QDs can be effectively increased with capping the ZnS shell. Meanwhile, the ultrafast dynamics including the hot carrier relaxation within hundreds of femtoseconds, carriers capture by defect states in tens of picoseconds, and electron-hole nonradiative recombination with several nanoseconds have been clearly identified for the as-prepared QDs with different shell thickness. We found that above carrier dynamics demonstrate an obvious dependence of ZnS shell thickness due to the effective isolation of the internal carriers in the CdxZn1-xSeyS1-y core from the decreased interfacial defects. These findings suggest that the capping shell can effectively regulate the carrier dynamics and blinking behavior, and improve the optical properties of CdxZn1-xSeyS1-y @ZnS QDs, which are significant for understanding the correlation between the structure and optical properties of fluorescent QDs.