Resolving Multi-Asperity Contacts at the Nanoscale through Super-Resolution Fluorescence Imaging.

Contact mechanics, spanning nanometer to tectonic scales, faces long-standing challenges arising from multiscale random roughness, which hinders experimental validation of theories. Understanding multi-asperity rough contacts is vital for addressing catastrophic consequences of these contacts failing such as earthquakes and for diverse technological applications. To visualize such contacts, we introduce a super-resolution microscopy method utilizing spontaneous millisecond ON/OFF fluorescence blinking of contact-sensitive molecular rotor molecules immobilized on a glass coverslip. This technique achieves ∼55 nm lateral imaging resolution for rough poly(methyl methacrylate) and glass spheres on glass contacts. For soft polymer spheres due to large plastic deformation, the resolution improvement does not significantly affect the area of real contact. However, for hard glass spheres, the real contact area is found to be 2.4 times smaller than that found by diffraction-limited imaging. This study highlights, through direct visualization, the impact of material stiffness on the nanoscale structure within the area of real contact.

Impact of Ring-Closing on the Photophysical Properties of One-Dimensional π-Conjugated Molecular Aggregate.

The self-assembled state of molecules plays a pivotal role in determining how inherent molecular properties transform and give rise to supramolecular functionalities and has long attracted attention. However, understanding the influence of morphologies spanning the nano- to mesoscopic scales of supramolecular assemblies derived from identical intermolecular interactions has been notoriously challenging due to dynamic structural change and monomer exchange of assemblies in solution. In this study, we demonstrate that curved one-dimensional molecular assemblies (supramolecular polymers) of lengths of around 70-200 nm, originating from the same luminescent molecule, exhibit distinct photoluminescent properties when they form closed circular structures (toroids) versus when they possess chain termini in solution (random coils). By exploiting the difference in kinetic stability between the toroids and random coils, we developed a dialysis protocol to selectively purify the former. It was revealed that these terminus-free closed structures manifest higher energy and more efficient luminescence compared with their mixed state with random coils. Time-resolved fluorescence measurements unveiled that random coils, due to their dynamic structural fluctuation in solution, generate local defects throughout the main chain, leading to luminescence from lower energy levels. In mixtures of the two assemblies, luminescence was exclusively observed from such a lower energy level of random coils, a result attributed to energy transfer between the assemblies. This work emphasizes that for identical supramolecular assemblies, only averaged properties have traditionally been considered, but their structures at the nano- to mesoscopic scale are important especially if they have a certain degree of shape persistency even in solution.

Steroid receptor coactivator TAIMAN is a new modulator of insect circadian clock.

TAIMAN (TAI), the only insect ortholog of mammalian Steroid Receptor Coactivators (SRCs), is a critical modulator of ecdysone and juvenile hormone (JH) signaling pathways, which govern insect development and reproduction. The modulatory effect is mediated by JH-dependent TAI's heterodimerization with JH receptor Methoprene-tolerant and association with the Ecdysone Receptor complex. Insect hormones regulate insect physiology and development in concert with abiotic cues, such as photo- and thermoperiod. Here we tested the effects of JH and ecdysone signaling on the circadian clock by a combination of microsurgical operations, application of hormones and hormone mimics, and gene knockdowns in the linden bug Pyrrhocoris apterus males. Silencing taiman by each of three non-overlapping double-strand RNA fragments dramatically slowed the free-running period (FRP) to 27-29 hours, contrasting to 24 hours in controls. To further corroborate TAIMAN's clock modulatory function in the insect circadian clock, we performed taiman knockdown in the cockroach Blattella germanica. Although Blattella and Pyrrhocoris lineages separated ~380 mya, B. germanica taiman silencing slowed the FRP by more than 2 hours, suggesting a conserved TAI clock function in (at least) some insect groups. Interestingly, the pace of the linden bug circadian clock was neither changed by blocking JH and ecdysone synthesis, by application of the hormones or their mimics nor by the knockdown of corresponding hormone receptors. Our results promote TAI as a new circadian clock modulator, a role described for the first time in insects. We speculate that TAI participation in the clock is congruent with the mammalian SRC-2 role in orchestrating metabolism and circadian rhythms, and that TAI/SRCs might be conserved components of the circadian clock in animals.

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.

Air-stable mixed cation lead halide perovskite films and microscopic study of their degradation process.

We report the preparation and nanoscale photophysical characterization of mixed cation perovskite films of the composition MA1-xFAxPbI3, with x = 0, 0.3 and 0.5. Films with x = 0.5 and 0.3 prepared in air using ethyl acetate as an antisolvent in a one-step spin-coating process are compositionally stable in ambient air for more than a year, in contrast to films prepared using a chlorobenzene antisolvent. The onset of degradation of the films near the film edges was monitored using in situ photoluminescence (PL) spectroscopy. The PL spectra of the degradation products are consistent with the PL spectra of 2D perovskite sheets of varying thicknesses. Morphologically, aging of the films brings about coalescing of the film grain structure into larger crystal grains. Furthermore, monitoring of the time traces of PL from individual nanoscale locations in the films (PL blinking) reveals that aging of the films does not change the extent of dynamic PL quenching or affect the observed long-range charge diffusion on the order of micrometers.

Simultaneous Force and Fluorescence Spectroscopy on Single Chains of Polyfluorene: Effect of Intra-Chain Aggregate Coupling.

Conjugated polymer chains in compact conformations or in films exhibit spectral features that can be attributed to interactions between individual conjugated segments of the chain, including formation of aggregates or excimers. Here, we use atomic force microscopy (AFM) on single chains of the conjugated polymer polyfluorene (PFO) to control the intersegment interactions by mechanically unfolding the chain. Simultaneously with the force spectroscopy we monitor fluorescence from the single PFO chains using a fluorescence microscope. We found that mechanical stretching of the chain causes disappearance of the green emission band. This observation provides evidence that the green emission originates from an intrachain aggregated state on the self-folded chain, which is decoupled by the stretching. In addition, the stretching upon laser irradiation leads to the appearance of additional features in the force spectra, small force peaks in the initial stages of the unfolding. These features are attributed to a combination of excitonic and van der Waals coupling of a ground-state intrachain aggregate.

Dynamic molecular conformational change leading to energy transfer in F8-5% BSP copolymer revealed by single-molecule spectroscopy.

Polyfluorene-based copolymers such as poly(9,9-dioctylfluorene)-alt-5% [bis-N,N'-(4-butylphenyl)-bis-N,N'-phenyl-1,4-phenylenediamine] (F8-5% BSP) are efficient blue-emitting polymers with various electronic phases: F8 blue-emitting glassy phase, F8 ordered more red-emitting ß-phase, and F8/BSP charge transfer (CT) state. Polymer light-emitting device performance and color purity can be significantly improved by forming ß-phase segments. However, the role of the ß-phase on energy transfer (ET) among glassy F8, ß-phase, and F8/BSP CT state is unclear. Herein, we identify dynamic molecular conformation-controlled ET from locally excited states to either the CT state or ß-phase in light-emitting copolymers. By conducting single-molecule spectroscopy for single F8-5% BSP chains, we find inefficient intra-chain ET from glassy segments to the CT state, while efficient ET from the glassy to the ß-phase. Spontaneous and reversible CT on-off emission is observed both in the presence and absence of the ß-phase. The density functional theory calculations reveal the origin of the on-chain CT state and indicate this CT emission on-off switching behavior could be related to molecule torsional motion between BSP and F8 units. The population of the CT state by ET can be increased via through-space interaction between the F8 block and the BSP unit on a self-folded chain. Temperature-dependent single-molecule spectroscopy confirms such interaction showing a gradual increase in intensity of the CT emission with the temperature. Based on these observations, we propose the dynamic molecular motion-induced conformation change as the origin of the glassy-to-CT ET, and thermal energy may provide the activation for such a change to enhance the ET from glassy or ß-phases to the CT state.

Cryptochrome-dependent magnetoreception in a heteropteran insect continues even after 24†h in darkness.

Sensitivity to magnetic fields is dependent on the intensity and color of light in several animal species. The light-dependent magnetoreception working model points to cryptochrome (Cry) as a protein cooperating with its co-factor flavin, which possibly becomes magnetically susceptible upon excitation by light. The type of Cry involved and what pair of magnetosensitive radicals are responsible is still elusive. Therefore, we developed a conditioning assay for the firebug Pyrrhocoris apterus, an insect species that possesses only the mammalian cryptochrome (Cry II). Here, using the engineered Cry II null mutant, we show that: (i) vertebrate-like Cry II is an essential component of the magnetoreception response, and (ii) magnetic conditioning continues even after 25 h in darkness. The light-dependent and dark-persisting magnetoreception based on Cry II may inspire new perspectives in magnetoreception and cryptochrome research.

Enhancement of the Photocurrent of a Single Photosystem I Complex by the Localized Plasmon of a Gold Nanorod.

A combination of conductive atomic force microscopy (AFM) and confocal fluorescence microscopy was used to measure photocurrents passing through single trimeric photosytem I (PSI) complexes located in the vicinity of single gold nanorods (AuNRs). Simultaneous excitation of PSI and of the AuNR longitudinal plasmon mode and detection of photocurrents from individual PSI in relation to the position of single AuNRs enable insight into plasmon-induced phenomena that are otherwise inaccessible in ensemble experiments. We have observed photocurrent enhancement by the localized plasmons by a factor of 2.9 on average, with maximum enhancement values of up to 8. Selective excitation of the longitudinal plasmon modes by the polarization of the excitation laser enables controllable switch-on of the photocurrent enhancement. The dependence of the extent of enhancement on the distance between PSI and AuNRs indicates that, apart from the enhancement of absorption, there is an additional enhancement mechanism affecting directly the electron transport process. The present study provides deeper insight into the molecular mechanisms of plasmon-enhanced photocurrents, not only in PSI but also potentially in other systems as well.

Dynamic Polymer Free Volume Monitored by Single-Molecule Spectroscopy of a Dual Fluorescent Flapping Dopant.

Single-molecule spectroscopy (SMS) of a dual fluorescent flapping molecular probe (N-FLAP) enabled real-time nanoscale monitoring of local free volume dynamics in polystyrenes. The SMS study was realized by structural improvement of a previously reported flapping molecule by nitrogen substitution, leading to increased brightness (22 times) of the probe. In a polystyrene thin film at the temperature of 5 K above the glass transition, the spectra of a single N-FLAP molecule undergo frequent jumps between short- and long-wavelength forms, the latter one indicating planarization of the molecule in the excited state. The observed spectral jumps were statistically analyzed to reveal the dynamics of the molecular environment. The analysis together with MD and QM/MM calculations show that the excited-state planarization of the flapping probe occurs only when sufficiently large polymer free volume of more than, at least, 280 Å3 is available close to the molecule, and that such free volume lasts for an average of 1.2 s.

Real-Time Monitoring of Formation and Dynamics of Intra- and Interchain Phases in Single Molecules of Polyfluorene.

Poly(9,9-dioctylfluorene) (PFO) is one of the most important conjugated polymer materials, exhibiting outstanding photophysical and electrical properties. PFO is also known for a diversity of morphological phases determined by conformational states of the main chain. Our goal in this work is to address some of the key questions on formation and dynamics of one such conformation, the ß-phase, by following in real time the evolution of fluorescence spectra of single PFO chains. The PFO is dispersed in a thin polystyrene film, and the spectra are monitored during the process of solvent vapor annealing with toluene. We confirm unambiguously that the PFO ß-phase segments are formed on a true single-chain level at room temperature in the solvent-softened polystyrene. We further find that the formation of the ß-phase is a dynamic and reversible process occurring on the order of seconds, leading to repeated spontaneous transitions between the glassy and ß-phase segments during the annealing. Comparison of PFO with two largely different molecular weights (Mw) shows that chains with lower Mw form the ß-phase segments much faster. For the high Mw PFO chains, a detailed Franck-Condon analysis of the ß-phase spectra shows a large distribution of the Huang-Rhys factor, S, and even dynamic changes of this factor occurring on a single chain. Such dynamics are likely a manifestation of changing coherence length of the exciton. Further, for the high Mw PFO chains we observe an additional conformational state, a crystalline γ-phase. The γ-phase formation is also a spontaneous reversible process in the solvent-softened matrix. The phase can form from both the ß-phase and the glassy phase, and the formation requires high Mw to enable intersegment interactions in a self-folded chain.

Toxicity of nanomaterials due to photochemical degradation and the release of heavy metal ions.

The increased production of semiconductor nanomaterials such as heavy metal quantum dots and perovskites for applications such as in energy harvesting, optoelectronic devices, bioanalysis, phototherapy and consumer health products raises concerns regarding nanotoxicity. After disposal, these materials degrade upon interaction with the environment, such as rain and surface waters, soil and oxygen, and solar irradiation, leading to the release of heavy metal ions in the environment with exposure to aquatic and terrestrial animals and plants, and humans. Researchers are in the early stages of understanding the potential toxicity of such nanomaterials by quantifying the amount of heavy metal ions released due to environmental or biological transformation. Here, we evaluate the toxicity of environmentally transformed nanomaterials by considering PbS quantum dots as a model system. Using metal ion sensors and steady-state fluorescence spectroscopy, we quantify the amount of Pb2+ released by the photochemical etching of quantum dots. Furthermore, with the help of cytotoxicity and comet assays, and DNA gel electrophoresis, we evaluate the adverse effects of the released metal ions into the cultured lung epithelial (H1650), and neuronal (PC12) cells. These studies reveal higher levels of cell proliferation and DNA damage to PC12 cells, suggesting the neurotoxicity of lead due to not only the downregulation of glutathione, elevated levels of reactive oxygen and nitrogen species, and a calcium influx but also the proactivation of activator protein 1 that is correlated with protein kinase c. This research shows the significance of molecular biology studies on different cells and animals to critically understand the health and environmental costs of heavy metal-based engineered nanomaterials.

Mechanically Induced Conformation Change, Fluorescence Modulation, and Mechanically Assisted Photodegradation in Single Nanoparticles of the Conjugated Polymer Poly(9,9-dioctylfluorene).

We explored the possibility of nanoscale mechanical manipulation and control of photophysical properties of conjugated polymer nanoparticles. We carried out a simultaneous atomic force microscopy (AFM) and fluorescence microspectroscopy study on single nanoparticles of the conjugated polymer poly(9,9-dioctylfluorene). The nanoparticles are prepared by a reprecipitation method and have an average height of 27 nm, and their emission is dominated by the well-ordered ß-phase conformation. Fluorescence polarization anisotropy and numerical simulations show that each particle contains at least three partly oriented straight ß-phase segments surrounded by amorphous glass-phase polyfluorene chains. In the simultaneous experiments, an AFM tip was used to apply external force on a single nanoparticle, and a confocal fluorescence microscope was used to monitor in real time the resulting changes in the fluorescence intensity and spectra. In a nitrogen atmosphere, weak to moderate force of up to 1 µN acts mainly on the glass-phase polyfluorene chains by forming quenchers that cause an efficient and reversible fluorescence decrease, whereas the ß-phase segments stay unaffected. A higher force of 5 µN, on the contrary, breaks the ß-phase segments into multiple glass-phase segments, causing a net increase in fluorescence intensity. Under ambient air conditions, even a moderate force of 1 µN strongly accelerates the degradation of the nanoparticle by preferably photobleaching the ß-phase and partially transforming it into the glass phase. These results will contribute to the fundamental knowledge on the relationship between photophysical and structural properties of polyfluorene nanostructures, and will also provide important feedback for potential applications of such nanostructures in flexible optoelectronic devices.

Toward accurate measurement of the intrinsic quantum yield of lanthanide complexes with back energy transfer.

Trivalent lanthanide complexes are an important class of luminescent material characterized by their strong absorption of light by the organic ligands and subsequent energy transfer to the lanthanide ion, realizing intense luminescence from the ion. With this mechanism of luminescence, the total quantum yield of a lanthanide complex is the product of the energy transfer efficiency from the ligand to the lanthanide ion and the "intrinsic" quantum yield of the lanthanide ion itself. The "absolute" method in measuring the quantum yield uses an integrating sphere, and this method can be used for measuring both the total and the intrinsic quantum yields. The presence of back energy transfer (the reverse process of energy transfer) adds complication to this by affecting both the dynamics of the excited state of the ligands and the lanthanide ion. Herein, we theoretically derive an equation that shows that in the presence of back energy transfer the intrinsic quantum yield may differ depending on whether it is determined from the measurement through excitation of the ligands or the lanthanide directly. The value measured by direct lanthanide excitation could decrease to 20% or less of the actual value when back energy transfer is prominent. Several previously reported Tb(iii) complexes are within the range to be cautious. This report shows that the "absolute" method for measuring the lanthanide ion-centered quantum yield may not be suitable in the presence of back energy transfer by principle. We also provide a possible workaround in the case that several approximations and assumptions can be made.

Single-particle electroluminescence of CsPbBr3 perovskite nanocrystals reveals particle-selective recombination and blinking as key efficiency factors.

Halide perovskites nanocrystals (NCs) are being explored as promising materials for optoelectronic applications, such as light-emitting devices or lasers. However, electroluminescence devices prepared from such NCs have long suffered from low efficiency and there has been no systematic study on the nanoscale origin of the poor efficiencies. Here, we use single-particle spectroscopy to compare electroluminescence and photoluminescence on the level of individual NCs of the perovskite CsPbBr3. The NCs form aggregates in a conducting matrix used as an emission layer in an electroluminescence device. In electroluminescence, only a small fraction of the NCs within the aggregate is emitting as a result of efficient charge migration, accumulation and selective recombination on larger NCs, leading to pronounced blinking and decreased efficiency. Under the condition of comparable excitation rates in both electroluminescence and photoluminescence, the intrinsic quantum yield in electroluminescence is on average 0.36 of that in photoluminescence.

Weak radiofrequency fields affect the insect circadian clock.

It is known that the circadian clock in Drosophila can be sensitive to static magnetic fields (MFs). Man-made radiofrequency (RF) electromagnetic fields have been shown to have effects on animal orientation responses at remarkably weak intensities in the nanotesla range. Here, we tested if weak broadband RF fields also affect the circadian rhythm of the German cockroach (Blatella germanica). We observed that static MFs slow down the cockroach clock rhythm under dim UV light, consistent with results on the Drosophila circadian clock. Remarkably, 300 times weaker RF fields likewise slowed down the cockroach clock in a near-zero static magnetic field. This demonstrates that the internal clock of organisms can be sensitive to weak RF fields, consequently opening the possibility of an influence of man-made RF fields on many clock-dependent events in living systems.

Photoinduced photoluminescence enhancement in self-assembled clusters of formamidinium lead bromide perovskite nanocrystals.

Nanocrystals of formamidinium lead bromide perovskite (FAPbBr3) self-assemble into clusters in powder and film samples and provide a prolonged photoluminescence lifetime, which is attributed to the diffusion of charge carriers through interparticle states formed among nanocrystals. Interestingly, the photoluminescence lifetime decreases and the emission intensity increases for the clusters, which is with the increase in the intensity of excitation light. By doping the nanocrystal clusters with C60, we successfully harvested the photogenerated charge carriers. Nonetheless, at high intensities of excitation, the rate of radiative recombination becomes comparable to that of the electron transfer to C60. Thus, the optimum rate of electron transfer to C60 is accomplished by minimally exciting the self-assembled nanocrystals.

Suppressed Triplet Exciton Diffusion Due to Small Orbital Overlap as a Key Design Factor for Ultralong-Lived Room-Temperature Phosphorescence in Molecular Crystals.

Persistent room-temperature phosphorescence (RTP) under ambient conditions is attracting attention due to its strong potential for applications in bioimaging, sensing, or optical recording. Molecular packing leading to a rigid crystalline structure that minimizes nonradiative pathways from triplet state is often investigated for efficient RTP. However, for complex conjugated systems a key strategy to suppress the nonradiative deactivation is not found yet. Here, the origin of small rates of a nonradiative decay process from triplet states of conjugated molecular crystals showing RTP is reported. Optical microscopy analysis showed that, despite a favorable molecular stacking, an aromatic crystal with strong RTP is characterized by small diffusion length and small values of the diffusion coefficient of triplet excitons. Quantum chemical calculations reveal a large overlap between the lowest unoccupied molecular orbitals but very small overlap between the highest occupied molecular orbitals (HOMOs). Inefficient electron exchange caused by the small overlap of HOMOs prevents triplet excitons from diffusing over long distances and consequently from quenching at defect sites inside the crystal or at the crystal surface. These results will allow design of comprehensive molecular structures to obtain molecular solids with more efficient RTP.

Stark Effect and Environment-Induced Modulation of Emission in Single Halide Perovskite Nanocrystals.

Organic-inorganic halide perovskites have emerged as promising materials for next-generation solar cells. In nanostructured form also, these materials are excellent candidates for optoelectronic applications such as lasers and light-emitting diodes for displays and lighting. While great progress has been achieved so far in optimizing the intrinsic photophysical properties of perovskite nanocrystals (NCs), in working optoelectronic devices, external factors, such as the effects of conducting environment and the applied electric field on exciton generation and photon emission, have been largely unexplored. Here, we use NCs of the all-inorganic perovskite CsPbBr3 dispersed polyvinyl carbazole, a hole-conductor, and in poly(methyl methacrylate), an insulator, to examine the effects of applied electric field and conductivity of the matrix on the perovskite photophysics at the single-particle level. We found that the conducting environment causes a significant decrease of photoluminescence (PL) brightness of individual NCs due the appearance of intermediate-intensity emitting states with significantly shortened lifetime. Applied electric field has a similar effect and, in addition, causes a nonlinear spectral shift of the PL maxima, a combination of linear and quadratic Stark effects caused by environment-induced polarity and field-related polarizability. The environment and electric-field effects are explained by ionization of the NCs through hole transfer and emission of the resulting negatively charged excitons.

Wavelength-Tunable Band-Edge Photoluminescence of Nonstoichiometric Ag-In-S Nanoparticles via Ga3+ Doping.

The nonstoichiometry of I-III-VI semiconductor nanoparticles, especially the ratio of group I to group III elements, has been utilized to control their physicochemical properties. We report the solution-phase synthesis of nonstoichiometric Ag-In-S and Ag-In-Ga-S nanoparticles and results of the investigation of their photoluminescence (PL) properties in relation to their chemical compositions. While stoichiometric AgInS2 nanoparticles simply exhibited only a broad PL band originating from defect sites in the particles, a narrow band edge PL peak newly appeared with a decrease in the Ag fraction in the nonstoichiometric Ag-In-S nanoparticles. The relative PL intensity of this band edge emission with respect to the defect-site emission was optimal at a Ag/(Ag + In) value of ca. 0.4. The peak wavelength of the band edge emission was tunable from 610 to 500 nm by increased doping with Ga3+ into Ag-In-S nanoparticles due to an increase of the energy gap. Furthermore, surface coating of Ga3+-doped Ag-In-S nanoparticles, that is, Ag-In-Ga-S nanoparticles, with a GaS x shell drastically and selectively suppressed the broad defect-site PL peak and, at the same time, led to an increase in the PL quantum yield (QY) of the band edge emission peak. The optimal PL QY was 28% for Ag-In-Ga-S@GaS x core-shell particles, with green band-edge emission at 530 nm and a full width at half-maximum of 181 meV (41 nm). The observed wavelength tunability of the band-edge PL peak will facilitate possible use of these toxic-element-free I-III-VI-based nanoparticles in a wide area of applications.