Time-resolved Mode Research Articles

Delineating the core and surface state heterogeneity of carbon dots during electron transfer.

Exploring the heterogeneity of carbon dots (C-Dots) is challenging because of the existence of complex structural diversity, and it is a demanding task for the development and designing of efficient C-Dots for various applications. Herein, we studied the role of the core state and surface state of C-Dots in heterogeneity via the successful investigation of the electron transfer (ET) process between different (blue, green, and red) emitting C-Dots and an electron acceptor methyl viologen (MV2+) using steady-state and time-resolved fluorescence and ultrafast transient absorption (TA) spectroscopic techniques. Selective excitation in the steady-state and time-resolved mode shows that the ET ability of the core state is higher than that of the surface state. Moreover, the kinetics of MV+˙ generation was probed using TA spectroscopy after the excitation of the core and surface state, where we observed that the surface state becomes less efficient due to the presence of an oxygen-containing functional group in the surface state, which acts as an electron scavenger. Moreover, the heterogeneity of the core and surface state was explored through the detection of the MV+˙ generation yield after the irradiation of UV and visible light (exciting the core and surface state). The result indicates that the graphitic nitrogen content in the core state and the oxygen-containing functional group in the surface state play an important role in the heterogeneity in the structure and the ET process. Our findings on the fundamental understanding of the heterogeneity of different emissive C-Dots will provide a new way of designing and developing a metal-free light-harvesting system.

(Invited) Electrochemistry Combined in Situ Neutron Reflectometry: A Powerful Tool for a More Sustainable Environment

Neutrons, having extraordinary characteristics that include penetrating power, sensitivity to lighter elements (such as hydrogen and carbon), and magnetic moment, make neutron reflectometry an ideal candidate for examining surface and interfacial structures of thin films in a non-destructive and non-invasive fashion. Liquids Reflectometer (LR) at the Spallation Neutron Source at Oak Ridge National Laboratory measures specular and off specular neutron reflectivity in a horizontal sample geometry, and tracks changes of layer thickness, scattering length density, and roughness as a function of depth. The exceptional sample environment at LR offers the capability to conduct in situ electrochemistry to probe the morphological and structural changes of solid/liquid interfaces during operation as a function of chemical and electrochemical gradients over time. Furthermore, the time-resolved mode at LR can be carried out continuously over a reduced Q range and binned into 60-second intervals. The improved time resolution of the neutron reflectivity (NR) measurement can help elucidate the kinetic behavior of surface/interfaces under dynamic conditions and provide a unique perspective to the equilibrium study. In this talk, I will give a brief introduction on neutron reflectometry, the LR, and a variety of sample environments at the LR. I will focus on the electrochemical applications coupled with in situ NR measurement. In the end, I will provide a specific example of applications in electrochemically modulated separation for water remediation by using neutron reflectometry. With the rapid growth of human population, it is highly desirable to minimize energy and materials input in selective separations for a more sustainable environment. The study is critical for creating new, more selective separation systems through rationally design of the structure and operating conditions.

Time-resolved fluorescence immunoassay based on glucose oxidase-encapsulated metal-organic framework for amplified detection of foodborne pathogen

BackgroundFluorescence immunoassays are commonly employed for the detection of pathogenic bacteria as a means of ensuring food safety and preserving public health. However, the challenges such as poor photostability and background interference have limited their sensitivity and accuracy. The emergence of metal-organic frameworks (MOFs) as a label probe offers a promising solution for advancing fluorescence immunoassays owing to their tunable nature. Nonetheless, the low fluorescence efficiency of MOFs and the potential risk of dye leakage pose obstacles to achieving high detection sensitivity. Therefore, there exists a pressing need to fully utilize the potential of MOF composites in fluorescence immunoassays. ResultsWe explored the potential of glucose oxidase-encapsulated zeolitic imidazole framework-90 (GOx@ZIF-90) as a label probe to construct a time-resolved fluorescence immunoassay with amplified detection signal. This immunoassay involved functionalizing Fe3O4 nanoparticle with porcine antibody to specifically capture and separate the target bacteria, Staphylococcus aureus (S. aureus). The captured S. aureus was then bound by GOx@ZIF-90 modified with vancomycin, resulting in a fluorescence response in the europium tetracycline (EuTc). The encapsulation of GOx in ZIF-90 provided a confinement effect that significantly enhanced the catalytic activity and stability of GOx. This led to a highly efficient conversion of glucose to H2O2, amplifying the fluorescence signal of EuTc. The immunoassay demonstrated a high sensitivity in detecting S. aureus, with a detection limit of 2 CFU/mL. We also obtained satisfactory results in milk samples. Attractively, the time-resolved detection mode of EuTc allowed the immunoassay to eliminate background fluorescence and enhance accuracy. SignificanceThis study not only presented a new method for detecting foodborne pathogens but also highlighted the potential of enzyme-encapsulated MOF composites as label probes in immunoassays, providing valuable insights for the design and fabrication of MOF composites for various applications.

A Switch-On Affinity-Based Iridium(III) Conjugate Probe for Imaging Mitochondrial Glutathione S-Transferase in Breast Cancer Cells.

Glutathione S-transferase is heterogeneously expressed in breast cancer cells and is therefore emerging as a potential diagnostic biomarker for studying the heterogeneity of breast cancers. However, available fluorescent probes for GSTs depend heavily on GSTs-catalyzed glutathione (GSH) nucleophilic substitution reactions, making them susceptible to interference by the high concentration of nucleophilic species in the cellular environment. Moreover, the functions of subcellular GSTs are generally overlooked due to the lack of suitable luminescence probes. Herein, we report a highly selective affinity-based luminescence probe 1 for GST in breast cancer cells through tethering a GST inhibitor, ethacrynic acid, to an iridium(III) complex. Compared to activity-based probes which require the use of GSH, this probe could image GST-pi in the mitochondria by directly adducting to GST-pi (or potentially GST-pi/GS) in living cells. Probe 1 possesses desirable photophysical properties including a lifetime of 911 ns, a Stokes shift of 343 nm, and high photostability. The "turn on" luminescence mode of the probe enables highly selective detection of the GST with a limit of detection of 1.01 μM, while its long emission lifetime allows sensitive detection in organic dye-spiked autofluorescence samples by a time-resolved mode. The probe was further applied to specifically and quantitatively visualize MDA-MB-231 cells via specific binding to mitochondrial GST, and could differentiate breast cell lines based on their expression levels of GST. To the best of our knowledge, this probe is the first affinity-based iridium(III) imaging probe for the subcellular GST. Our work provides a valuable tool for unmasking the diverse roles of a subcellular GST in living systems, as well as for studying the heterogeneity of breast cancers.

The Analytical Application of Quenching Phenomena of CdTe Quantum Dot Nanoparticles

This paper is devoted to the synthesis and application of CdTe quantum dot (QD) nanoparticles covered with organic ligands containing a thiol group, mostly mercaptopropionic acid (MPA) and glutathione (GSH). The simple one-step synthetic procedure was optimized to prepare greater quantities of nanoparticles for analytical purposes. The prepared CdTe QD nanoparticles were characterized by various analytical techniques, and their interaction with some metal ions (Cu(II), Pb(II), and Hg(II)) was studied by using luminescence spectroscopy in both steady-state and time-resolved modes. The mathematical analysis of the quenching effect of Cu(II) ions on the luminescence of CdTe QD nanoparticles shows that the static contribution is mostly responsible for the overall effect, but experimental conditions, such as pH, ionic strength, or the concentration of nanoparticles in aqueous solution, could also be important. The presence of metal ions in the form of a metal complex species could play an important role, and this phenomenon could be used to tune the selectivity of the quenching process. These findings have been utilized for the development of an analytical procedure for the detection and quantitative analysis of Cu(II) and Pb(II) ions in environmental water samples. In practice, this procedure could be easily implemented in a microplate format to increase throughput.

Radiotherapy delivery error detection with electronic portal imaging device (EPID)-based in vivo dosimetry.

Radiotherapy delivery error detection with electronic portal imaging device (EPID)-based in vivo dosimetry.

A beamline for ultrafast extreme ultraviolet magneto-optical spectroscopy in reflection near the shot noise limit.

High harmonic generation (HHG) makes it possible to measure spin and charge dynamics in materials on femtosecond to attosecond timescales. However, the extreme nonlinear nature of the high harmonic process means that intensity fluctuations can limit measurement sensitivity. Here we present a noise-canceled, tabletop high harmonic beamline for time-resolved reflection mode spectroscopy of magnetic materials. We use a reference spectrometer to independently normalize the intensity fluctuations of each harmonic order and eliminate long term drift, allowing us to make spectroscopic measurements near the shot noise limit. These improvements allow us to significantly reduce the integration time required for high signal-to-noise (SNR) measurements of element-specific spin dynamics. Looking forward, improvements in the HHG flux, optical coatings, and grating design can further reduce the acquisition time for high SNR measurements by 1-2 orders of magnitude, enabling dramatically improved sensitivity to spin, charge, and phonon dynamics in magnetic materials.

Coherent Flow Structures Near Tongue Region in a Centrifugal Fan With Forward-Curved Blades

Abstract The unsteady characteristics of the velocity field around the tongue of the volute in a centrifugal fan with forward-curved blades were investigated by time-resolved particle image velocimetry and mode decomposition analysis, i.e., proper orthogonal decomposition (POD) and spectral proper orthogonal decomposition (SPOD). Both POD and SPOD analysis reveal the existence of two types of unsteadiness in the flow field in the volute, one is the large-scale fluctuations at rotation frequency and its high-order harmonics in the main outlet flow, the other is due to the jet–wake structures at blade passing frequency in the cutoff clearance region. Additionally, SPOD analysis reveals a third type of disturbance, which is characterized as strip-like velocity structures at the intermediate frequency. The geometric and dynamic features of these coherent flow structures are inspected by the eigenfunction and their reduced-order reconstruction. In comparison with the POD analysis, the SPOD analysis enables the examination of the spatial pattern of each frequency component due to its dual-orthogonality nature. These structures direct impact the tongue, and thus might be responsible for the generation of strong wall-pressure fluctuation on the nose of the tongue. Due to the frequency mixing limitation of POD analysis, they cannot be easily identified from the leading-order POD modes. Furthermore, the frequency range of these three groups of disturbances, as well as their spatial extension, are determined via SPOD analysis and reduced-order reconstruction.

Prospects of Genetically Encoded Flim Indicators for the Quantitative Assessment of Intracellular Parameters

A significant number of genetically encoded indicators based on fluorescent proteins that allow detecting changes in various parameters: membrane potential shift, pH, concentrations of hydrogen peroxide, lactate, pyruvate, NAD+/NADH, ATP, calcium cations, etc. have been created. Some of them (for example, indicators of calcium cations and hydrogen peroxide) are successfully used by numerous groups of researchers in experiments in vivo. The fundamental limitation of most fluorescent indicators is their ability to only qualitatively assess the change in the parameter of interest in the cell. The nature of this limitation is that it is difficult to compare the absolute value of the measured parameter and the detected fluorescent signal for various reasons. Quantitative measurements impose an additional requirement on the indicator signal: it should not depend on the technical features of the experiment (such as the level of protein expression, the type and power of the excitation source, a set of fluorescent filters, etc.). The fulfillment of this requirement may be associated with the detection of the indicator signal in the time domain, where the main characteristic of the signal is the fluorescence lifetime. The fluorescence lifetime is an absolute value, insensitive to changes in the intensity of the excitation source, detection settings, losses in the optical path or sample, changes in the concentration of the fluorophore in the sample, photobleaching. Thus, when calibrating an indicator in the time domain, a specific value of the measured parameter is associated with a specific value of the fluorescence lifetime. The time-resolved imaging mode imposes its own limitations on the properties of the indicator's fluorescent core. Thus, the lifetime of its fluorescence should change in proportion to the value of the measured parameter, which is not characteristic of any fluorophore. In this work, we make a brief review of the principles of detection of the fluorescent signal of genetically encoded indicators, focusing on current approaches related to the analysis of time-resolved data.

Surface and Structure of Phosphatidylcholine Membranes Reconstructed with CoFe2O4 Nanoparticles.

Structural changes in phosphatidylcholine lipid membranes caused by the introduction of insoluble CoFe2O4 nanoparticles (NPs) are analyzed. Changes in nuclear magnetic resonance spectrum, infrared spectrum, and ionic conductivity of membranes are observed with the addition of NPs. The presence of NPs in membranes is proved by atomic force and magnetic force microscopy. Structural changes in the membranes in the vicinity of the lipid C-O bonds caused by NPs are observed by Scanning near-field optical microscopy. Analysis of nuclear magnetic resonance (NMR) spectra allowed us to identify the affected atomic groups in the membrane surface layers. Conductivity measurements of the bilayer membranes were performed in DC as well as in time-resolved modes. Hydrophobic NPs stimulate surface distortion and creation of pores, which depending on NP concentration leads to an increase in the ionic conductivity of membranes. Concentration dependence demonstrating percolation threshold was analyzed in the frame of the fractal theory approach.

Time-resolved mode analysis in Er-doped self-sweeping ring fiber laser

We present here a detailed study of the mode composition of an Er-doped ring self-sweeping fiber laser. Time-resolved heterodyne measurements prove that the radiation consists of slightly overlapping rectangular mode packets, each of which corresponds to generation of individual longitudinal modes. Optical frequency of each following packet sequentially increases by two free spectral ranges of the laser cavity. The presence of parametric processes during sequential growth of the neighboring mode packets is experimentally demonstrated for the first time in a self-sweeping laser. Instantaneous linewidth of the laser radiation does not exceed 4 kHz during every step of single longitudinal mode generation, which is also accompanied by a small change of the optical frequency (chirp) of ∼40 kHz associated with an integral change of the laser cavity refractive index. The results are useful for calculations and modeling of laser systems based on dynamic gratings.

Single-cell and bulk ICP-MS investigation of accumulation patterns of Pt-based metallodrugs in cisplatin-sensitive and -resistant cell models.

In research enabling preclinical development and attaining a deeper understanding of the behavior of metallodrugs in cancer cells with acquired resistance, intracellular Pt accumulation could be considered an important biomarker and analytical focus. In this work, Pt accumulation patterns in terms of the number of cells and Pt mass in single cells were precisely defined by using inductively coupled plasma-mass spectrometry (ICP-MS) operating in a fast time-resolved analysis mode. This technique is otherwise known as single-cell (SC)-ICP-MS. By applying the nascent and validated SC-ICP-MS technique, comparisons across three Pt drugs (cisplatin, carboplatin, and oxaliplatin) in the A2780 and A2780cis ovarian cancer cell models could be made. Additional roles of transporters on top of passive diffusion and the drugs' bioactivity could be postulated. The SC-ICP-MS-based observations also served as a cross-validation point to augment preexisting research findings on Pt-resistance mechanisms. Conjectures regarding S and Fe metabolism were also derived based on an additional and direct ICP-MS analysis of endogenous elements. Overall, our work not only confirms the utility of SC-ICP-MS in chemotherapeutic research, but also provided insights into further ICP-MS-based analytical capacities to be developed.

Light Scattering and Turbidimetry Techniques for the Characterization of Nanoparticles and Nanostructured Networks.

Light scattering and turbidimetry techniques are classical tools for characterizing the dynamics and structure of single nanoparticles or nanostructured networks. They work by analyzing, as a function of time (Dynamic Light Scattering, DLS) or angles (Static Light Scattering, SLS), the light scattered by a sample, or measuring, as a function of the wavelength, the intensity scattered over the entire solid angle when the sample is illuminated with white light (Multi Wavelength Turbidimetry, MWT). Light scattering methods probe different length scales, in the ranges of (DLS), or (Wide Angle SLS), or (Low Angle SLS), and some of them can be operated in a time-resolved mode, with the possibility of characterizing not only stationary, but also aggregating, polymerizing, or self-assembling samples. Thus, the combined use of these techniques represents a powerful approach for studying systems characterized by very different length scales. In this work, we will review some typical applications of these methods, ranging from the field of colloidal fractal aggregation to the polymerization of biologic networks made of randomly entangled nanosized fibers. We will also discuss the opportunity of combining together different scattering techniques, emphasizing the advantages of a global analysis with respect to single-methods data processing.

On the Dynamics of Transient Plasmas Generated by Nanosecond Laser Ablation of Several Metals.

The dynamics of transient plasma generated by UV ns-laser ablation of selected metals (Co, Cu, Ag, Bi) were investigated by the Langmuir Probe method in angle- and time-resolved modes. Multiple ionic and electronic structures were seen for all plasmas with some corresponding to anions or nanoparticle-dominated structures. The addition of an Ar atmosphere energetically confined the plasma and increased the charge density by several orders of magnitude. For pressure ranges exceeding 0.5 Pa fast ions were generated in the plasma as a result of Ar ionization and acceleration in the double layer defining the front of the plasma plume. Several correlations between the target nature plasma properties were attempted. The individual plasma structure expansion velocity increases with the melting point and decreases with the atomic mass while the corresponding charged particle densities decrease with the melting point, evidencing the relationship between the volatility of the sample and the overall abated mass.

A Novel Approach for the Determination of the Ge Isotope Ratio Using Liquid-Liquid Extraction and Hydride Generation by Multicollector Inductively Coupled Plasma Mass Spectrometry.

In this work, a method for the accurate and precise determination of the Ge isotope ratio in synthetic water and natural samples of geological origin using multicollector inductively coupled plasma mass spectrometry (MC-ICPMS) with hydride generation was developed. The method was based on the liquid–liquid extraction of Ge to eliminate all elements affecting the generation of germanium hydrides. The standard-sample bracketing method was used to correct instrumental bias. Registration of analytical signal in time-resolved mode gave way to choose signals with best parameters and improved the precision of the results. Controlling the pH by using acetic buffer boosted the sensitivity by nearly five times in comparison to hydride generation methods suggested by other authors. The newly developed method is much simpler and quicker, does not need laborious Ge separation with ion-exchange resins, and thanks to its superior sensitivity, allows measurements of the Ge isotopic ratio in materials with relatively low Ge content. Delta values of the 74Ge/70Ge isotope ratio were measured in standard reference materials for which reference values were available in the GeoREM database. We demonstrated that the accuracy and precession of this method are equally good or better than methods proposed by other authors.

Singlet oxygen luminescence detector based on low-cost InGaAs avalanche photodiode

Molecular oxygen excited to singlet state (Singlet oxygen, 1O2) becomes highly reactive and cytotoxic chemical. 1O2 is commonly generated by photoexcitation of dyes (photosensitizers), including the photodynamic therapy and diagnostics of cancer. However, the formation of singlet oxygen is often unwanted for various light-sensitive compounds, e.g. it causes the photobleaching of fluorescent probes. In either case, during a development of new photosensitive chemicals and drugs there is a need to evaluate the amount of 1O2 formed during photoexcitation. The direct approach in measuring the amount of singlet oxygen is based on the detection of its luminescence at 1270 nm. However, this luminescence is usually weak, which implies the use of highly sensitive single-photon detectors. Thus the existing instruments are commonly complicated and expensive. Here we suggest an approach and report a device to measure the 1O2 luminescence using low-cost InGaAs avalanche photodiode and simple electronics. The measurements can be performed in stationary (not time-resolved) mode in organic solvents such as tetrachloromethane (CCl4), ethanol and DMSO. In particular, we performed spectral-resolved measurements of the singlet oxygen luminescence in CCl4 with the device and demonstrated high complementarity to literature data. The simple setup allows to evaluate the efficiency (or speed) of singlet oxygen generation and hence facilitates the development and characterization of new photosensitizers and other photosensitive chemicals.

Time-resolved spectral-domain optical coherence tomography with CMOS SPAD sensors.

We present a first spectral-domain optical coherence tomography (SD-OCT) system deploying a complementary metal-oxide-semiconductor (CMOS) single-photon avalanche diode (SPAD) based, time-resolved line sensor. The sensor with 1024 pixels achieves a sensitivity of 87 dB at an A-scan rate of 1 kHz using a supercontinuum laser source with a repetition rate of 20 MHz, 38 nm bandwidth, and 2 mW power at 850 nm centre wavelength. In the time-resolved mode of the sensor, the system combines low-coherence interferometry (LCI) and massively parallel time-resolved single-photon counting to control the detection of interference spectra on the single-photon level based on the time-of-arrival of photons. For proof of concept demonstration of the combined detection scheme we show the acquisition of time-resolved interference spectra and the reconstruction of OCT images from selected time bins. Then, we exemplify the temporal discrimination feature with 50 ps time resolution and 249 ps timing uncertainty by removing unwanted reflections from along the optical path at a 30 mm distance from the sample. The current limitations of the proposed technique in terms of sensor parameters are analysed and potential improvements are identified for advanced photonic applications.

Volume and compressibility differences between protein conformations revealed by high-pressure NMR

Proteins often interconvert between different conformations in ways critical to their function. Although manipulating such equilibria for biophysical study is often challenging, the application of pressure is a potential route to achieve such control by favoring the population of lower volume states. Here, we use this feature to study the interconversion of ARNT PAS-B Y456T, which undergoes a dramatic +3 slip in the β-strand register as it switches between two stably folded conformations. Using high-pressure biomolecular NMR approaches, we obtained the first, to our knowledge, quantitative data testing two key hypotheses of this process: the slipped conformation is both smaller and less compressible than the wild-type equivalent, and the interconversion proceeds through a chiefly unfolded intermediate state. Data collected in steady-state pressure and time-resolved pressure-jump modes, including observed pressure-dependent changes in the populations of the two conformers and increased rate of interconversion between conformers, support both hypotheses. Our work exemplifies how these approaches, which can be generally applied to protein conformational switches, can provide unique information that is not easily accessible through other techniques.

Acquisition of kHz-frequency two-dimensional surface temperature field using phosphor thermometry and proper orthogonal decomposition assisted long short-term memory neural networks

A reconstruction technique of kHz time-resolved two-dimensional (2D) surface temperature field was achieved with the discrete point measurements and low sampling rate 2D thermographic phosphor (TP) thermometry measurements using a long short-term memory (LSTM) based artificial intelligence framework. The 2D surface temperature field of a 350 °C plate with a 2.5 Hz swing cooling jet was measured using TP thermometry at a sampling rate of 20 Hz. At the same time, high-frequency thermocouples with a sampling rate of 1 kHz were recorded for the construction of LSTM neural networks training and for validation. The 20 Hz 2D surface temperature field was analyzed with proper orthogonal decomposition to acquire the energy modes and model coefficients. The mode coefficients are then trained with the discrete but high-frequency time-resolved temperature information from the thermocouple by LSTM to acquire the time-resolved mode coefficients. Finally, the high-frequency time-resolved 2D surface temperature field is obtained by reconstructing modes and the time-resolved coefficients. The reconstructed result shows that the current technique can obtain high time-resolved and spatially resolved 2D surface temperature fields very well.

Quantitative Elemental Analysis of a Single Cell by Using Inductively Coupled Plasma-Mass Spectrometry in Fast Time-Resolved Analysis Mode.

The elemental composition of a single yeast, green alga, or red blood cell (RBC) was precisely determined by using inductively coupled plasma-mass spectrometry (ICP-MS) operating in fast time-resolved analysis (TRA) mode. The technique is known as single-cell (SC)-ICP-MS. Phosphorus, sulfur, magnesium, zinc, and iron were detected in the three types of cell. The elemental composition of yeast and green alga obtained by SC-ICP-MS was consistent with results obtained from conventional ICP-MS measurements following acid digestion of the cells. Slight differences were found in the measured values between SC-ICP-MS and the conventional ICP-MS results for RBC. However, the SC-ICP-MS results for S and Fe in RBC were closer to the estimated values for these elements that were calculated from the level of hemoglobin in RBCs. The data suggest that SC-ICP-MS is suitable for the analysis of various cell types, namely, fungus, plant, and animal cells.