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Time-resolved flow cytometry (TRFC) was used to measure metabolic differences in estrogen receptor-positive breast cancer cells. This specialty cytometry technique measures fluorescence lifetimes as a single-cell parameter thereby providing a unique approach for high-throughput cell counting and screening. Differences in fluorescence lifetime were detected and this was associated with sensitivity to the commonly prescribed therapeutic tamoxifen. Differences in fluorescence lifetime are attributed to the binding states of the autofluorescent metabolite NAD(P)H. The function of NAD(P)H is well described and in general involves cycling from a reduced to oxidized state to facilitate electron transport for the conversion of pyruvate to lactate. NAD(P)H fluorescence lifetimes depend on the bound or unbound state of the metabolite, which also relates to metabolic transitions between oxidative phosphorylation and glycolysis. To determine if fundamental metabolic profiles differ for cells that are sensitive to tamoxifen compared to those that are resistant, large populations of MCF-7 breast cancer cells were screened and fluorescence lifetimes were quantified. Additionally, metabolic differences associated with tamoxifen sensitivity were measured with a Seahorse HS mini metabolic analyzer (Agilent Technologies Inc. Santa Clara, CA) and confocal imaging. Results show that tamoxifen-resistant breast cancer cells have increased utilization of glycolysis for energy production compared to tamoxifen-sensitive breast cancer cells. This work is impacting because it establishes an early step toward developing a reliable screening technology in which large cell censuses can be differentiated for drug sensitivity in a label-free fashion. [ABSTRACT FROM AUTHOR]

This work proposes MP‐Grasp4D (magnetization‐prepared golden‐angle radial sparse parallel 4D) MRI, a free‐breathing, inversion recovery (IR)‐prepared, time‐resolved 4D MRI technique with improved T1‐weighted contrast. MP‐Grasp4D MRI acquisition incorporates IR preparation into a radial gradient echo sequence. MP‐Grasp4D employs a golden‐angle navi‐stack‐of‐stars sampling scheme, where imaging data of rotating radial stacks and navigator stacks (acquired at a consistent rotation angle) are alternately acquired. The navigator stacks are used to estimate a temporal basis for low‐rank subspace‐constrained reconstruction. This allows for the simultaneous capture of both IR‐induced contrast changes and respiratory motion. One temporal frame of the imaging volume in MP‐Grasp4D MRI is reconstructed from a single stack and an adjacent navigator stack on average, resulting in a nominal temporal resolution of 0.16 seconds per volume. Images corresponding to the optimal inversion time (TI) can be retrospectively selected for providing the best image contrast. Reader studies were conducted to assess the performance of MP‐Grasp4D MRI in liver imaging across 30 subjects in comparison with standard Grasp4D MRI without IR preparation. MP‐Grasp4D MRI received significantly higher scores (P < 0.05) than Grasp4D in all assessment categories. There was a moderate to almost perfect agreement (kappa coefficient from 0.42 to 0.9) between the two readers for image quality assessment. When the scan time is reduced, MP‐Grasp4D MRI preserves image contrast and quality, demonstrating additional acceleration capability. MP‐Grasp4D MRI improves T1‐weighted contrast for free‐breathing time‐resolved 4D MRI and eliminates the need for explicit motion compensation. This method is expected to be valuable in different MRI applications such as MR‐guided radiotherapy. [ABSTRACT FROM AUTHOR]

Premise: Plants generate a wide array of signals such as olfactory cues to attract and manipulate the response of pollinators. The present study addresses the temporal patterns of scent emission as an additional dimension to the scent composition. The expectation is that divergent floral function is reflected in divergent qualitative and temporal emission patterns. Methods: We used GC‐ion mobility spectrometry with an integrated pre‐concentration for automated acquisition of the temporal trends in floral volatile emissions for N. viridiflorus, N. papyraceus, and N. cantabricus subsp. foliosus. Results: We found a considerable increase in scent emissions and changes in scent composition for N. viridiflorus at night. This increase was particularly pronounced for aromatic substances such as benzyl acetate and p‐cresol. We found no diurnal patterns in N. papyraceus, despite a similar qualitative composition of floral volatiles. Narcissus cantabricus subsp. foliosus showed no diurnal patterns either and differed considerably in floral scent composition. Conclusions: Scent composition, circadian emission patterns, and floral morphology indicate divergent, but partially overlapping pollinator communities. However, the limited pollinator data from the field only permits a tentative correlation between emission patterns and flower visitors. Narcissus papyraceus and N. cantabricus show no clear diurnal patterns and thus no adjustment to the activity patterns of their diurnal pollinators. In N. viridiflorus, timing of scent emission indicates an adaptation to nocturnal flower visitors, contradicting Macroglossum as the only reported pollinator. We propose that the legitimate pollinators of N. viridiflorus are nocturnal and are still unidentified. [ABSTRACT FROM AUTHOR]

A profound understanding of the binding dynamics of drugs with plasma proteins is highly important as it determines its therapeutical potential. Herein, we have investigated the molecular interaction of Tucatinib (TUC), a novel, primitive and highly selective drug for treatment of HER2+ advanced unresectable breast cancer, with bovine serum albumin (BSA) using multi‐pronged spectroscopic techniques and molecular docking. TUC considerably quenched the intrinsic fluorescence of BSA via static quenching mechanism, and it binds to the hydrophobic cleft of subdomain IIA of BSA with a moderate binding affinity. The thermodynamic analysis revealed the major interplay of van der Waals forces and hydrogen bonding interactions in stabilising the binding. The Förster distance 'r' signifies feasible energy transfer between BSA and TUC. The conformational analyses indicate some alterations in the protein secondary structure as a consequence of some changes in the protein polar environment due to complex formation. The experimental results are supported by docking analyses. [ABSTRACT FROM AUTHOR]

Serial crystallography, born from groundbreaking experiments at the Linac Coherent Light Source in 2009, has evolved into a pivotal technique in structural biology. Initially pioneered at X‐ray free‐electron laser facilities, it has now expanded to synchrotron‐radiation facilities globally, with dedicated experimental stations enhancing its accessibility. This review gives an overview of current developments in serial crystallography, emphasizing recent results in time‐resolved crystallography, and discussing challenges and shortcomings. [ABSTRACT FROM AUTHOR]

Over 50 years of development, synchrotron based X-ray microscopy has become a routine and powerful tool for the analysis of nanoscale structure and chemistry in many areas of science. Scanning X-ray microscopy is particularly well suited to the study of chemical and magnetic states of matter and has become available at most synchrotron light sources using a variety of optical schemes, detectors and sample environments. The Advanced Light Source at Lawrence Berkeley National Laboratory has an extensive program of soft X-ray scanning microscopy which supports a broad range of scientific research using a suite of advanced tools for high spatio-temporal resolution and control of active materials. Instruments operating within an energy range between 200–2500 eV with spatial resolution down to 7 nm and sub 20 picosecond time resolution are available. These capabilities can be routinely used in combination with a variety of sample stimuli, including gas or fluid flow, temperature control from 100 to 1200 K, DC bias and pulsed or continuous microwave excitation. We present here a complete survey of our instruments, their most advanced capabilities and a perspective on how they complement each other to solve complex problems in energy, materials and environmental science.

Throughout history, the influenza A virus has caused numerous devastating global pandemics. Macrophages, as pivotal innate immune cells, exhibit a wide range of immune functions characterized by distinct polarization states, reflecting their intricate heterogeneity. In this study, we employed the time-resolved single-cell sequencing technique coupled with metabolic RNA labelling to elucidate the dynamic transcriptional changes in distinct polarized states of bone marrow-derived macrophages (BMDMs) upon infection with the influenza A virus. Our approach not only captures the temporal dimension of transcriptional activity, which is lacking in conventional scRNA-seq methods, but also reveals that M2-polarized Arg1_macrophage cluster is the sole state supporting successful replication of influenza A virus. Furthermore, we identified distinct antigen presentation capabilities to CD4+ T and CD8+ T cells across diverse polarized states of macrophages. Notably, the M1 phenotype, exhibited by (BMDMs) and murine alveolar macrophages (AMs), demonstrated superior conventional and cross-presentation abilities for exogenous antigens, with a particular emphasis on cross-presentation capacity. Additionally, as CD8+ T cell differentiation progressed, M1 polarization exhibited an enhanced capacity for cross-presentation. All three phenotypes of BMDMs, including M1, demonstrated robust presentation to CD4+ regulatory T cells, while displaying limited ability to present to naive CD4+ T cells. These findings offer novel insights into the immunological regulatory mechanisms governing distinct polarized states of macrophages, particularly their roles in restricting the replication of influenza A virus and modulating antigen-specific T cell responses through innate immunity.

Understanding the thermoelastic response of a nanostructure is crucial for the choice of materials and interfaces in electronic devices with improved and tailored transport properties at the nanoscale. Here, we show how the deposition of a MoS2 monolayer can strongly modify the nanoscale thermoelastic dynamics of silicon substrates close to their interface. We demonstrate this by creating a transient grating with extreme ultraviolet light, using ultrashort free-electron laser pulses, whose ≈84 nm period is comparable to the size of elements typically used in nanodevices, such as electric contacts and nanowires. The thermoelastic response, featuring coherent acoustic waves and incoherent relaxation, is tangibly modified by the presence of monolayer MoS2. Namely, we observed a major reduction of the amplitude of the surface mode, which is almost suppressed, while the longitudinal mode is basically unperturbed, aside from a faster decay of the acoustic modulations. We interpret this behavior as a selective modification of the surface elasticity, and we discuss the conditions to observe such effect, which may be of immediate relevance for the design of Si-based nanoscale devices. [ABSTRACT FROM AUTHOR]

The binding of drugs to plasma proteins determines its fate within the physiological system, hence profound understanding of its interaction within the bloodstream is important to understand its pharmacodynamics and pharmacokinetics and thereby its therapeutic potential. In this regard, our work delineates the mechanism of interaction of Selumetinib (SEL), a potent anti‐cancer drug showing excellent effect against multiple solid tumors, with plasma protein bovine serum albumin (BSA), using methods such as absorption, steady‐state fluorescence, time‐resolved, fluorescence resonance energy transfer, Fourier transform infrared spectra (FTIR), circular dichroism (CD), synchronous and 3D‐fluorescence, salt fluorescence, molecular docking and molecular dynamic simulations. The BSA fluorescence intensity was quenched with increasing concentration of SEL which indicates interactions of SEL with BSA. Stern–Volmer quenching analysis and lifetime studies indicate the involvement of dynamic quenching. However, some contributions from the static quenching mechanism could not be ruled out unambiguously. The association constant was found to be 5.34 × 105 M−1 and it has a single binding site. The Förster distance (r) indicated probable energy transmission between the BSA and SEL. The positive entropy changes and enthalpy change indicate that the main interacting forces are hydrophobic forces, also evidenced by the results of molecular modeling studies. Conformation change in protein framework was revealed from FTIR, synchronous and 3D fluorescence and CD studies. Competitive binding experiments as well as docking studies suggest that SEL attaches itself to site I (subdomain IIA) of BSA where warfarin binds. Molecular dynamic simulations indicate the stability of the SEL–BSA complex. The association energy between BSA and SEL is affected in the presence of different metals differently. [ABSTRACT FROM AUTHOR]

Due to its strong affinity for chlorine gas, Cd can potentially form CdCl2. Cd-doped CuO thin films are sensitive to chlorine gas, as careful inclusion of Cd in the CuO crystal structure modifies the energy gap depending on the dopant concentration. We employed spray pyrolysis to deposit Cd-doped CuO on fluorine tin oxide (FTO) while heating the substrate to 500 °C. The X-ray diffraction (XRD) analysis revealed that Cd was interstitially incorporated in the CuO lattice structure, as verified by scanning electron microscopy (SEM) imaging. The photoluminescence study demonstrated that increasing the Cd concentration in CuO resulted in higher emission intensity, providing valuable insights into Cu2+ and O2- energy levels. Exposing a Cd-doped CuO thin film to chlorine gas modifies the bandgap, depending on the Cd concentration. The fluctuation in the bandgap energy of copper oxide doped with cadmium indicates the chlorine gas concentration nearby. Time-resolved measurements for the I-V characteristics of the thin film revealed considerable current variation during the exposure to chlorine gas. [ABSTRACT FROM AUTHOR]

Significance: Hyperspectral time-resolved (TR) near-infrared spectroscopy offers the potential to monitor cytochrome-c-oxidase (oxCCO) and blood oxygenation in the adult brain with minimal scalp/skull contamination. We introduce a hyperspectral TR spectrometer that uses compressive sensing to minimize acquisition time without compromising spectral range or resolution and demonstrate oxCCO and blood oxygenation monitoring in deep tissue. Aim: Develop a hyperspectral TR compressive sensing spectrometer and use it to monitor oxCCO and blood oxygenation in deep tissue. Approach: Homogeneous tissue-mimicking phantom experiments were conducted to confirm the spectrometer's sensitivity to oxCCO and blood oxygenation. Twolayer phantoms were used to evaluate the spectrometer's sensitivity to oxCCO and blood oxygenation in the bottom layer through a 10 mm thick static top layer. Results: The spectrometer was sensitive to oxCCO and blood oxygenation changes in the bottom layer of the two-layer phantoms, as confirmed by concomitant measurements acquired directly from the bottom layer. Measures of oxCCO and blood oxygenation by the spectrometer were highly correlated with "gold standard" measures in the homogeneous and two-layer phantom experiments. Conclusions: The results show that the hyperspectral TR compressive sensing spectrometer is sensitive to changes in oxCCO and blood oxygenation in deep tissue through a thick static top layer. [ABSTRACT FROM AUTHOR]

Gas-phase anions present an ideal playground for the exploration of excited-state dynamics. They offer control in terms of the mass, extent of solvation, internal temperature, and conformation. The application of a range of ion sources has opened the field to a vast array of anionic systems whose dynamics are important in areas ranging from biology to star formation. Here, we review recent experimental developments in the field of anion photodynamics, demonstrating the detailed insight into photodynamical and electron-capture processes that can be uncovered. We consider the electronic and nuclear ultrafast dynamics of electronically bound excited states along entire reaction coordinates; electronically unbound states showing that photochemical concepts, such as chromophores and Kasha's rule, are transferable to electron-driven chemistry; and nonvalence states that straddle the interface between bound and unbound states. Finally, we consider likely developments that are sure to keep the field of anion dynamics buoyant and impactful. [ABSTRACT FROM AUTHOR]

Abstract Despite the recent rise of RNA-seq datasets combining single-cell (sc) resolution with 4-thiouridine (4sU) labelling, analytical methods exploiting their power to dissect transcriptional bursting are lacking. Here, we present a mathematical model and Bayesian inference implementation to facilitate genome-wide joint parameter estimation and confidence quantification (R package: burstMCMC). We demonstrate that, unlike conventional scRNA-seq, 4sU scRNA-seq resolves temporal parameters and furthermore boosts inference of dimensionless parameters via a synergy between single-cell resolution and 4sU labelling. We apply our method to published 4sU scRNA-seq data and linked with ChIP-seq data, we uncover previously obscured associations between different parameters and histone modifications.

ABSTRACTOver the last 20 years, XUV and X-ray free-electron lasers have enabled a wide variety of time-resolved experiments that have dramatically advanced our understanding of ultrafast molecular dynamics on atomic length scales and femtosecond time scales. This review focuses on experimental studies of ultrafast dynamics of atoms and molecules in the gas phase, tracing the development of the field from early proof-of-principle studies to recent pump-probe experiments that elucidate the coupled electronic and nuclear dynamics during photochemical reactions with a temporal resolution that is now extending into the attosecond domain.

The L-lactic acid (L-LA) fermentation process, based on sodium hydroxide neutralization, demonstrates environmental friendliness during product extraction. However, lactate fermentation is hindered by the pronounced stress effect of sodium lactate on the strain compared with calcium lactate. In this study, we performed time-resolved transcriptomic and proteomic analyses of Heyndrickxia coagulans DSM1 during NaOH-buffered L-LA production. The expression levels of the glycolytic genes demonstrated an initial increase followed by a subsequent decrease, whereas the tricarboxylic acid cycle genes exhibited an initial decrease followed by a subsequent increase throughout the fermentation process. Moreover, we identified clusters of genes consisting of transcription factors and ATP-binding cassette (ABC) transporters that demonstrate a progressive elevation of expression levels throughout the fermentation process, with significant upregulation observed at later stages. This investigation yields valuable insights into the response mechanisms of H. coagulans during NaOH-buffered L-LA fermentation and presents potential targets for metabolic engineering.

In this study, plasma induced by a nanosecond Nd:YAG laser on thin oil films deposited on a silica wafer was characterized by evaluating the main plasma parameters. Spatially and temporally integrated spectral measurements were performed under experimental conditions optimized for elemental analysis of trace metals in oil. Time-resolved values of the spectral line intensities, electron number density, and plasma temperature were obtained from time-integrated measurements by subtracting averaged spectra recorded at different time delays. The electron number density was estimated using the Stark broadened profile of the hydrogen Balmer alpha line. Ionization temperatures were derived from Mg ionic to atomic line intensity ratios. The obtained apparent values of time-resolved plasma parameters were in the range of 1.1×1017 cm-3 (1.5 μs) to 1.5×1016 cm-3 (4 μs) and 9400 K (3 μs) to 7200 K (5 μs), depending on the delay time. Emission spectra of C2 and CN molecules were used to evaluate the rotational and vibrational temperature.

Demand for low carbon energy storage has highlighted the importance of imaging techniques for the characterization of electrode microstructures to determine key parameters associated with battery manufacture, operation, degradation, and failure both for next generation lithium and other novel battery systems. Here, recent progress and literature highlights from magnetic resonance, neutron, X‐ray, focused ion beam, scanning and transmission electron microscopy are summarized. Two major trends are identified: First, the use of multi‐modal microscopy in a correlative fashion, providing contrast modes spanning length‐ and time‐scales, and second, the application of machine learning to guide data collection and analysis, recognizing the role of these tools in evaluating large data streams from increasingly sophisticated imaging experiments. [ABSTRACT FROM AUTHOR]

X‐ray tomography is a versatile tool in materials research and engineering since it allows for a non‐destructive and three‐dimensional mapping of the constituents of a heterogeneous material as long as they differ in their interactions with X‐rays. Recent developments of the technique have brought down the time needed for the acquisition of a single tomogram by many orders of magnitude compared to what was needed 25 years ago. Nowadays, up to 1000 full tomograms can be recorded in a second, which enables real‐time studies of changes in samples caused by reactions or by applied processing operations. The term tomoscopy has been coined for such sequences of 3D images. We review the application of X‐ray tomography and tomoscopy on metals and describe each step required and the associated challenges. A selection of representative investigations is presented with a focus on time‐resolved phenomena in metals and alloys ranging from mechanical deformation, solidification to metals processing processes such as welding and additive manufacturing. Finally likely future developments are discussed. [ABSTRACT FROM AUTHOR]

In this study, we investigated the enhanced charge transfer (CT) behaviors of single-walled carbon nanotubes (f-SWCNTs) and copper oxide (CuO)-reinforced poly(o-anisidine-co-o-toluidine) nanocomposites. Prior this study, the targeted nanocomposite materials were fabricated using an in situ oxidative polymerization tool. FT-IR, XRD, and SEM techniques were used to confirm the formation of the pure POA-co-POT copolymer and its related nanocomposites in the presence of f-SWCNTs and CuO nanoparticles. The enhancement of the thermal behavior and conductivity in the presence of either f-SWCNTs or f-SWCNTs/CuO confirms the well-dispersed filler on the polymeric matrix. Additionally, steady-state and time-resolved measurements were performed to monitor the influence of f-SWCNTs and CuO on the optical properties and CT related to the copolymerization process. Our fabricated materials showed a quenching of 25% and 31% in the presence of 10% f-SWCNTs, and 2% f-SWCNTs + 15% CuO, respectively. These findings provided strong evidence for the fast charge transfer within the fabricated nanocomposite, which matched the time-resolved results. The fs measurements showed that the ground state bleach (GSB) kinetics was more rapid in the presence of 10% f-SWCNTs compared to that of the free copolymer, while in the presence of 10% CuO, the electron transfer (ET) was faster, which decayed rapidly with a 0.6 ps time constant supporting the metallic nature of the composite. Our findings suggested that the incorporation of f-SWCNTs and CuO into a polymer matrix can close the gap resulting from polymer limitation. [ABSTRACT FROM AUTHOR]

Introduction: This study evaluated our experience with dynamic computed tomography angiography (dCTA) as a diagnostic tool after endovascular aortic aneurysm repair (EVAR) with respect to the endoleak classification and the available literature. Methods: We reviewed all patients who underwent dCTA because of suspected endoleaks after the EVAR and classified the endoleaks in these patients based on standard CTA (sCTA) and dCTA. We systematically reviewed all available publications that investigated the diagnostic accuracy of dCTA compared with other imaging techniques. Results: In our single-center series, 16 dCTAs were performed in 16 patients. In 11 patients, the undefined endoleaks that appeared on sCTA scans were successfully classified using dCTA. In three patients with a type II endoleak and aneurysm sac growth, inflow arteries were successfully identified using dCTA, and in two patients, aneurysm sac growth was observed without a visible endoleak on both sCTA and dCTA scans. The dCTA revealed four occult endoleaks, all of which were type II endoleaks. The systematic review identified six series comparing dCTA with other imaging methods. All articles reported an excellent outcome regarding the endoleak classification. In published dCTA protocols, the number and timing of phases varied greatly, affecting radiation exposure. Time attenuation curves of the current series show that some phases do not contribute to the endoleak classification and that the use of a test bolus improves the timing of the dCTA. Conclusions: The dCTA is a valuable additional tool that can identify and classify endoleaks more accurately than the sCTA. Published dCTA protocols vary greatly and should be optimized to decrease radiation exposure as long as accuracy can be maintained. The use of a test bolus to improve the timing of the dCTA is recommended, but the optimal number of scanning phases is yet to be determined. [ABSTRACT FROM AUTHOR]

Dynamic processes in catalysis are gaining increased attention and could very well be one of the next frontiers in surface science. One way to study such processes is to induce chemical changes on the surface for example by periodically adjusting the (electro)chemical potential in situ and identify the resulting spectral changes. Often this is referred to as Modulation Excitation Spectroscopy (MES). Using Ambient Pressure Photoelectron Spectroscopy data, we here discuss and compare the analysis of MES data using both Phase Sensitive Detection (PSD) and Fourier analysis. We discuss that PSD determines the component magnitude at a user-defined phase value while Fourier analysis provides the maximum oscillation amplitude and respective phase value of oscillating spectral features. We discuss advantages and disadvantages of the different analysis schemes and explore how the full time-evolution can be obtained. [Display omitted] • Phase Sensitive Detection and Fourier analysis are compared for the first time • Pros and cons and limitations of the PSD and Fourier analysis are discussed in detail • Fourier analysis enables on-the-fly detection of oscillating minority species in situ • For the special case of APXPS this enables structure-activity correlation • We hope our work will inspire many future studies of oscillating surface structures [ABSTRACT FROM AUTHOR]

The design and evaluation experiments of a multibeam X-ray tomography optical system that can be used with synchrotron radiation from sources with a narrow energy bandwidth, i.e. undulator sources, are reported. It consists of silicon single crystals that diffract the incident X-rays to 27 beams, which are used to image a sample. The energy of the beams was aligned with an accuracy sufficient for use at typical undulator beamlines. Projection images of a test sample were collected and successfully reconstructed, showing the feasibility of a high-speed X-ray tomography instrument based on the optical system.

We report on SwissSPAD2, an image sensor with 512×512 photon-counting pixels, each comprising a single-photon avalanche diode (SPAD), a 1-bit memory, and a gating mechanism capable of turning the SPAD on and off, with a skew of 250ps and 344ps, respectively, for a minimum duration of 5.75ns. The sensor is designed to achieve a frame rate of up to 97,700 binary frames per second and sub-40ps gate shifts. By synchronizing it with a pulsed laser and using multiple successive overlapping gates, one can reconstruct a molecule's fluorescent response with picosecond temporal resolution. Thanks to the sensor's number of pixels (the largest to date) and the fully integrated gated operation, SwissSPAD2 enables widefield FLIM with an all-solid-state solution and at relatively high frame rates. This was demonstrated with preliminary results on organic dyes and semiconductor quantum dots using both decay fitting and phasor analysis. Furthermore, pixels with an exceptionally low dark count rate and high photon detection probability enable uniform and high quality imaging of biologically relevant fluorescent samples stained with multiple dyes. While future versions will feature the addition of microlenses and optimize firmware speed, our results open the way to low-cost alternatives to commercially available scientific time-resolved imagers.

We report on SwissSPAD2, an image sensor with 512×512 photon-counting pixels, each comprising a single-photon avalanche diode (SPAD), a 1-bit memory, and a gating mechanism capable of turning the SPAD on and off, with a skew of 250ps and 344ps, respectively, for a minimum duration of 5.75ns. The sensor is designed to achieve a frame rate of up to 97,700 binary frames per second and sub-40ps gate shifts. By synchronizing it with a pulsed laser and using multiple successive overlapping gates, one can reconstruct a molecule's fluorescent response with picosecond temporal resolution. Thanks to the sensor's number of pixels (the largest to date) and the fully integrated gated operation, SwissSPAD2 enables widefield FLIM with an all-solid-state solution and at relatively high frame rates. This was demonstrated with preliminary results on organic dyes and semiconductor quantum dots using both decay fitting and phasor analysis. Furthermore, pixels with an exceptionally low dark count rate and high photon detection probability enable uniform and high quality imaging of biologically relevant fluorescent samples stained with multiple dyes. While future versions will feature the addition of microlenses and optimize firmware speed, our results open the way to low-cost alternatives to commercially available scientific time-resolved imagers.

Electron spectroscopy with the unprecedented transmission of angle-resolved time-of-flight detection, in combination with pulsed X-ray sources, brings new impetus to functional materials science. We showcase recent developments towards chemical sensitivity from electron spectroscopy for chemical analysis and structural information from photoelectron diffraction using the phase transition properties of 1T-TaS2. Our development platform is the SurfaceDynamics instrument located at the Femtoslicing facility at BESSY II, where femtosecond and picosecond X-ray pulses can be generated and extracted. The scientific potential is put into perspective to the current rapidly developing pulsed X-ray source capabilities from Lasers and Free-Electron Lasers. [ABSTRACT FROM AUTHOR]

Detecting electromagnetic radiation scattered from a tip-sample junction has enabled overcoming the diffraction limit and started the flourishing field of polariton nanoimaging. However, most techniques only resolve amplitude and relative phase of the scattered radiation. Here, we utilize field-resolved detection of ultrashort scattered pulses to map the dynamics of surface polaritons in both space and time. Plasmon polaritons in graphene serve as an ideal model system for the study, demonstrating how propagating modes can be visualized and modeled in the time domain by a straightforward mathematical equation and normalization method. This novel approach enables a direct assessment of the polaritons' group and phase velocities, as well as the damping. Additionally, it is particularly powerful in combination with a pump-probe scheme to trace subcycle changes in the polariton propagation upon photoexcitation. Our method readily applies to other quantum materials, providing a versatile tool to study ultrafast nonequilibrium spatiotemporal dynamics of polaritons.

The interaction between macromolecular proteins and small molecule ligands is an essential component of cellular function. Such ligands may include enzyme substrates, molecules involved in cellular signalling or pharmaceutical drugs. Together with biophysical techniques used to assess the thermodynamic and kinetic properties of ligand binding to proteins, methodology to determine high-resolution structures that enable atomic level interactions between protein and ligand(s) to be directly visualised is required. Whilst such structural approaches are well established with high throughput X-ray crystallography routinely used in the pharmaceutical sector, they provide only a static view of the complex. Recent advances in X-ray structural biology methods offer several new possibilities that can examine protein-ligand complexes at ambient temperature rather than under cryogenic conditions, enable transient binding sites and interactions to be characterised using time-resolved approaches and combine spectroscopic measurements from the same crystal that the structures themselves are determined. This Perspective reviews several recent developments in these areas and discusses new possibilities for applications of these advanced methodologies to transform our understanding of protein-ligand interactions.

Small-angle X-ray scattering (SAXS) is a method for examining the solution structure, oligomeric state, conformational changes, and flexibility of biomacromolecules at a scale ranging from a few Angstroms to hundreds of nanometers. Wide time scales ranging from real time (milliseconds) to minutes can be also covered by SAXS. With many advantages, SAXS has been extensively used, it is widely used in the structural characterization of biomacromolecules in food science and technology. However, the application of SAXS in charactering the structure of food biomacromolecules has not been reviewed so far. In the current review, the principle, theoretical calculations and modeling programs are summarized, technical advances in the experimental setups and corresponding applications of in situ capabilities: combination of chromatography, time-resolved, temperature, pressure, flow-through are elaborated. Recent applications of SAXS for monitoring structural properties of biomacromolecules in food including protein, carbohydrate and lipid are also highlighted, and limitations and prospects for developing SAXS based on facility upgraded and artificial intelligence to study the structural properties of biomacromolecules are finally discussed. Future research should focus on extending machine time, simplifying SAXS data treatment, optimizing modeling methods in order to achieve an integrated structural biology based on SAXS as a practical tool for investigating the structure-function relationship of biomacromolecules in food industry.

The electrochemistry and photophysics of the Pt(II) complexes [Pt(naphen)(X)] (Hnaphen = naphtho[1,2-b][1,10]phenanthroline, X = Cl or C≡CPh) containing the rigid tridentate C^N^N-coordinating pericyclic naphen ligand was studied alongside the complexes of the tetrahydro-derivative [Pt(thnaphen)(X)] (Hthnaphen = 5,6,8,9-tetrahydro-naphtho[1,2-b][1,10]phenanthroline) and the N^C^N-coordinated complex [Pt(bdq)(Cl)] (Hbdq = benzo[1,2-h:5,4-h']diquinoline. The cyclic voltammetry showed reversible reductions for the C^N^N complexes, with markedly fewer negative potentials (around −1.6 V vs. ferrocene) for the complexes containing the naphen ligand compared with the thnaphen derivatives (around −1.9 V). With irreversible oxidations at around +0.3 V for all of the complexes, the naphen made a difference in the electrochemical gap of about 0.3 eV (1.9 vs. 2.2 eV) compared with thnaphen. The bdq complex was completely different, with an irreversible reduction at around −2 V caused by the N^C^N coordination pattern, which lacked a good electron acceptor such as the phenanthroline unit in the C^N^N ligand naphen. Long-wavelength UV-Vis absorption bands were found around 520 to 530 nm for the C^N^N complexes with the C≡CPh coligand and were red-shifted when compared with the Cl derivatives. The N^C^N-coordinated bdq complex was markedly blue-shifted (493 nm). The steady-state photoluminescence spectra showed poorly structured emission bands peaking at around 630 nm for the two naphen complexes and 570 nm for the thnaphen derivatives. The bdq complex showed a pronounced vibrational structure and an emission maximum at 586 nm. Assuming mixed 3LC/3MLCT excited states, the vibronic progression for the N^C^N bdq complex indicated a higher LC character than assumed for the C^N^N-coordinated naphen and thnaphen complexes. The blue-shift was a result of the different N^C^N vs. C^N^N coordination. The photoluminescence lifetimes and quantum yields ΦL massively increased from solutions at 298 K (0.06 to 0.24) to glassy frozen matrices at 77 K (0.80 to 0.95). The nanosecond time-resolved study on [Pt(naphen)(Cl)] showed a phosphorescence emission signal originating from the mixed 3LC/3MLCT with an emission lifetime of around 3 µs. [ABSTRACT FROM AUTHOR]

An experimental investigation was performed on the wake flow field of an isolated roughness element of a flat plate at Mach 6 by employing the nanoparticle-based planar laser scattering (NPLS) approach. The three-dimensional features and causes of the flow field structure were scrutinized by transient flow field images of roughness elements on various planes. The time-resolved NPLS technique was implemented to examine the time evolution characteristics of the wake flow field of roughness elements. In the following, the process of dynamic evolution of large-scale vortex structures in the wake flow field was methodically assessed. Additionally, the influences of roughness element heights on the wake vortex structure were evaluated and the obtained results were compared. [ABSTRACT FROM AUTHOR]

Enzymes, as a kind of biomacromolecules, play an important role in many physiological processes and relate directly to various diseases. Developing an efficient detection method for enzyme activity is important to achieve early diagnosis of enzyme‐relevant diseases and high throughput screening of potential enzyme‐relevant drugs. Time‐resolved luminescence assay provide a high accuracy and signal‐to‐noise ratios detection methods for enzyme activity, which has been widely used in high throughput screening of enzyme‐relevant drugs and diagnosis of enzyme‐relevant diseases. Inspired by these advantages, various responsive probes based on metal complexes and metal‐free organic compounds have been developed for time‐resolved bioimaging and biosensing of enzyme activity owing to their long luminescence lifetimes, high quantum yields and photostability. In this review, we comprehensively reviewed metal complex‐ and metal‐free organic compound‐based responsive probes applied to detect enzyme activity through time‐resolved imaging, including their design strategies and sensing principles. Current challenges and future prospects in this rapidly growing field are also discussed. [ABSTRACT FROM AUTHOR]

Local pH near the catalyst surface exerts a substantial influence on both catalytic selectivity and activity in electrocatalytic CO 2 reduction (CO 2 RR). While the investigation of local pH remains challenging in the aspect of in-situ imaging and monitoring. In this study, a novel lateral-type surface-enhanced Raman spectroscopy (L-SERS) system with a wide pH testing range from acidic to alkaline, is developed for the in-situ observation of local pH distributions with high spatial and temporal resolution during CO 2 RR. Excellent in-situ imaging of local pH near the catalyst surface is obtained, showing good consistency with simulation results. Besides, the gradual pH elevation and stabilization can be monitored via this time-resolved local pH investigation. This study presents a direct visualization of local pH distribution and variation at the Nernst layer during CO 2 RR, providing direct evidence in local pH for the feasibility of acidic electrolyte and offering theoretical guidance for optimizing CO 2 RR conditions. [Display omitted] • A novel lateral-type SERS system to in-situ observe local pH is developed for CO 2 RR with a wide pH testing range. • Excellent in-situ imaging of local pH is achieved and the gradual varying process of local pH is monitored. • Results provide a direct evidence in local pH for the feasibility of acidic electrolyte in CO 2 RR. [ABSTRACT FROM AUTHOR]

• Synchrotron-based operando APXPS extended to the steady-growth regime of ALD. • Ligand-exchange mechanism in ALD of TiO 2 from TTIP/H 2 O spectroscopically confirmed. • Spectroscopic evidence for a static role of the TiO 2 surface in the TTIP/H 2 O process. Today, atomic layer deposition (ALD) has become a firm corner stone of thin film deposition technology. The microelectronics industry, an early adopter of ALD, imposes stringent requirements on ALD to produce films with highly defined physical and chemical properties, which becomes even more important as device and component dimensions decrease. This, in turn, means that our understanding of the chemical processes underlying ALD needs to increase exponentially. Here, we show that one can use synchrotron-based time-resolved ambient pressure x-ray photoelectron spectroscopy (APXPS) to obtain highly detailed operando information on the surface chemistry of ALD, not only, as proven earlier, during the initial ALD cycles, but also for the steady-growth regime reached during the later stages of deposition. Using event averaging and Fourier-transform methods, we show that the ALD of TiO 2 from titanium tetraisopropoxide (TTIP) and water precursors in the steady-growth regime follows the suggested ligand-exchange reaction mechanism, with no sign of oxygen transport between the deposited layers and the bulk of the film, as has been observed for other materials systems. Hence, the TiO 2 ALD from TTIP and water constitutes a textbook example of metal oxide ALD, as expected for this well-known ALD process. The detailed insight is made possible by computerised control of the precursor pulses that enable the recording of long data sets, which comprise many ALD cycles at highly regular intervals, in combination with an advanced data analysis that allows us to pick out signals undetectable in the raw data. The analysis method also allows to separate oscillating contributions to the signals induced by the ALD pulsing from the overwhelming bulk signal. [Display omitted] [ABSTRACT FROM AUTHOR]

Despite its simplicity, Boltzmann thermometry suffers from limited relative sensitivities since the energy gap must be in the order of magnitude of the thermal energy used for probing. Luminescent crossover thermometry is a potential alternative to exploit high energy gaps at low temperatures based on a generally high non-radiative coupling rate via a configurational crossover. The number of possible candidates for this type of temperature-dependent luminescence is quite versatile. Inorganic systems include transition metal ions with split d orbitals in a ligand field such as Cr3+ and Mn4+, mercury-like s2 ions such as Bi3+ or Pb2+ with coupled 3P0 and 3P1 levels, or lanthanoid ions with low energetic 4fn-15d1 levels next to their 4fn spin-orbit levels. In the case of organic emitters, thermally activated delayed fluorescence (TADF) has become increasingly popular, which relies on thermal coupling between an excited singlet and triplet state via intersystem crossing. While the general concept of two thermally excited radiatively emitting states is valid for each of the mentioned types of luminophores and thus, thermometry is in principle possible, the mechanistic details of the thermal coupling can be different. In this tutorial and theoretically motivated article, I will shortly review the relevant aspects of the excited state kinetics of two thermally coupled levels and demonstrate how the ratio between radiative and intrinsic non-radiative coupling rates affects the possible readout choices for optical signals used as a measure for temperature and how this influences the relative sensitivity of ratiometric and lifetime-based thermometers. An overview over various possible inorganic and organic emitters with the potential for thermometry will be given and based on these kinetic considerations, it will be assessed what readout mode is most recommendable for a given luminescent center.