Ms Time Resolution Research Articles

Subsecond-Resolved Molecular Measurements in the Living Body Using Chronoamperometrically Interrogated Aptamer-Based Sensors.

Electrochemical, aptamer-based (E-AB) sensors support the continuous, real-time measurement of specific small molecules directly in situ in the living body over the course of many hours. They achieve this by employing binding-induced conformational changes to alter electron transfer from a redox-reporter-modified, electrode-attached aptamer. Previously we have used voltammetry (cyclic, alternating current, and square wave) to monitor this binding-induced change in transfer kinetics indirectly. Here, however, we demonstrate the potential advantages of employing chronoamperometry to measure the change in kinetics directly. In this approach target concentration is reported via changes in the lifetime of the exponential current decay seen when the sensor is subjected to a potential step. Because the lifetime of this decay is independent of its amplitude (e.g., insensitive to variations in the number of aptamer probes on the electrode), chronoamperometrically interrogated E-AB sensors are calibration-free and resistant to drift. Chronoamperometric measurements can also be performed in a few hundred milliseconds, improving the previous few-second time resolution of E-AB sensing by an order of magnitude. To illustrate the potential value of the approach we demonstrate here the calibration-free measurement of the drug tobramycin in situ in the living body with 300 ms time resolution and unprecedented, few-percent precision in the determination of its pharmacokinetic phases.

Time resolved measurements of hydrogen ion energy distributions in a pulsed 2.45 GHz microwave plasma

A plasma diagnostic study of the Ion Energy Distribution Functions (IEDFs) of H+, H2+, and H3+ ions in a 2.45 GHz hydrogen plasma reactor called TIPS is presented. The measurements are conducted by using a Plasma Ion Mass Spectrometer with an energy sector and a quadrupole detector from HIDEN Analytical Limited in order to select an ion species and to measure its energy distribution. The reactor is operated in the pulsed mode at 100 Hz with a duty cycle of 10% (1 ms pulse width). The IEDFs of H+, H2+, and H3+ are obtained each 5 μs with 1 μs time resolution throughout the entire pulse. The temporal evolution of the plasma potential and ion temperature of H+ is derived from the data. It is shown that the plasma potential is within the range of 15–20 V, while the ion temperature reaches values of 0.25–1 eV during the pulse and exhibits a fast transient peak when the microwave radiation is switched off. Finally, the ion temperatures are used to predict the transverse thermal emittance of a proton beam extracted from 2.45 GHz microwave discharges.

Separation of Silver Nanoparticles with Different Coatings by Capillary Electrophoresis Coupled to ICP-MS in Single Particle Mode.

The possibility of separating mixtures of Ag nanoparticles (NPs) with similar sizes but different surface coatings using capillary electrophoresis coupled to single particle inductively coupled mass-spectrometry (CE-SP-ICP-MS) was investigated. In two-component mixtures, it was possible to separate 40 nm sized polyvinylpirrolidone (PVP)- and citrate-coated NPs, 40 nm sized polyethylene glycol (PEG)- and citrate-coated NPs, and 60 nm sized PVP- and citrate-coated NPs. The separation of a more complex mixture containing NPs with the different coatings and sizes was successful, and each component, namely, 20, 40, and 60 nm sized citrate-coated and 40 and 60 nm sized PVP-coated NPs, could be distinguished. The theoretically expected migration order was confirmed by experimental results with selected Ag NPs. On the basis of the experimental observations, a separation mechanism that considers the effect of stable vs displaceable coatings during NP migration in CE is suggested. The ICP-MS was equipped with a prototype data acquisition system (μsDAQ) that provided 5 μs time resolution.

High-Speed "4D" Computational Microscopy of Bacterial Surface Motility.

Bacteria exhibit surface motility modes that play pivotal roles in early-stage biofilm community development, such as type IV pili-driven "twitching" motility and flagellum-driven "spinning" and "swarming" motility. Appendage-driven motility is controlled by molecular motors, and analysis of surface motility behavior is complicated by its inherently 3D nature, the speed of which is too fast for confocal microscopy to capture. Here, we combine electromagnetic field computation and statistical image analysis to generate 3D movies close to a surface at 5 ms time resolution using conventional inverted microscopes. We treat each bacterial cell as a spherocylindrical lens and use finite element modeling to solve Maxwell's equations and compute the diffracted light intensities associated with different angular orientations of the bacterium relative to the surface. By performing cross-correlation calculations between measured 2D microscopy images and a library of computed light intensities, we demonstrate that near-surface 3D movies of Pseudomonas aeruginosa translational and rotational motion are possible at high temporal resolution. Comparison between computational reconstructions and detailed hydrodynamic calculations reveals that P. aeruginosa act like low Reynolds number spinning tops with unstable orbits, driven by a flagellum motor with a torque output of ∼2 pN μm. Interestingly, our analysis reveals that P. aeruginosa can undergo complex flagellum-driven dynamical behavior, including precession, nutation, and an unexpected taxonomy of surface motility mechanisms, including upright-spinning bacteria that diffuse laterally across the surface, and horizontal bacteria that follow helicoidal trajectories and exhibit superdiffusive movements parallel to the surface.

Implementation of Online Preconcentration and Microsecond Time Resolution to Capillary Electrophoresis Single Particle Inductively Coupled Plasma Mass Spectrometry (CE-SP-ICP-MS) and Its Application in Silver Nanoparticle Analysis.

Capillary electrophoresis (CE) coupled to single particle inductively coupled plasma mass-spectrometry (SP-ICP-MS) was used for the first time with a prototype data acquisition (μsDAQ) system that features 5 μs time resolution (100% duty cycle) to separate and quantify mixtures of silver nanoparticles (Ag NPs). Additionally, an online preconcentration technique, reversed electrode polarity stacking mode (REPSM), was applied for Ag NPs analysis with CE-SP-ICP-MS for the first time. After optimization, best results were achieved using a injection time of 110 s and a constant pressure of 50 mbar in hydrodynamic injection mode. It was possible to detect 14.3 ± 1.5× more 20 nm sized, 21.0 ± 4.2× more 40 nm sized, and 27.7 ± 4.9× more 60 nm sized Ag NPs compared to the standard injection time of only 3 s. The effect of applied voltage on the NPs separation was studied, and a CE separation at 20 kV was found to be optimal for the present setup. The capability of CE-SP-ICP-MS for quantification of particle number concentration was investigated, and detection limits in the submicrogram-per-liter range were achieved. The possibility to separate 20, 40, and 60 nm sized Ag NPs simultaneously present in a mixture was demonstrated over a broad concentration range.

Time‐resolved ultrasonic spectroscopy for bubbles

We show that ultrasound can provide time‐resolved measurements of the size distribution and the concentration of bubbles in a liquid. The potential of the technique is demonstrated by following disappearance of bubbles having an average radius of 20 μm with a 10 ms time resolution. We show that our technique can detect small concentrations of bubbles, with a large spectrum of accessible bubble radii (from 80 nm to 40 μm for a gas volume fraction of ), and with a sub‐millisecond time resolution. This new technique could be a valuable tool for investigating rapid processes such as nucleation or dissolution of bubbles. © 2017 American Institute of Chemical Engineers AIChE J, 63: 4666–4672, 2017

Highly Heterogeneous Nature of the Native and Unfolded States of the B Domain of Protein A Revealed by Two-Dimensional Fluorescence Lifetime Correlation Spectroscopy

Elucidating the protein folding mechanism is crucial to understand how proteins acquire their unique structures to realize various biological functions. With this aim, the folding/unfolding of small globular proteins has been extensively studied. Interestingly, recent studies have revealed that even such small proteins represent considerably complex processes. In this study, we examined the folding/unfolding process of a small α-helical protein, the B domain of protein A (BdpA), at equilibrium using two-dimensional fluorescence lifetime correlation spectroscopy with 10 μs time resolution. The results showed that although the BdpA is a two-state folder, both the native and unfolded states are highly heterogeneous and the conformational conversion within each ensemble occurs within 10 μs. Furthermore, it was shown that the average structures of both ensembles gradually change and become more elongated as the denaturant concentration increases. The analysis on two mutants suggested that fraying of the N-terminal helix is the origin of the inhomogeneity of the native state. Because the direct observation of the ensemble nature of the native state at the single-molecule level has not been reported, the data obtained in this study give new insights into complex conformational properties of small proteins.

Temporal evolution of two-dimensional electron temperature and ion flux on a substrate in a pulsed-power inductively coupled plasma

The two-dimensional temporal behavior of plasma was investigated at the wafer-level when applied power was turned on/off, which is one cycle of the pulsed plasma. For the measurement of transient plasma, the floating harmonic method and a post-processing method were used to obtain the ion flux and the electron temperature with a 1 ms time resolution. The results show that the transient behaviors of plasma such as antenna power absorption are observed in pulsed inductively coupled plasma. Antenna-shaped ion flux is observed when the power is turned on, and the radial profiles of the ion flux are followed by a diffusion solution when the power is turned off. Furthermore, the electron temperature shows pulsed plasma characteristics.

Development of hard X-ray spectrometer with high time resolution on the J-TEXT tokamak

A hard X-ray (HXR) spectrometer has been developed to study the runaway electrons during the sawtooth activities and during the runaway current plateau phase on the J-TEXT tokamak. The spectrometer system contains four NaI scintillator detectors and a multi-channel analyzer (MCA) with 0.5ms time resolution. The dedicated peak detection circuit embedded in the MCA provides a pulse height analysis at count rate up to 1.2 million counts per second (Mcps), which is the key to reach the high time resolution. The accuracy and reliability of the system have been verified by comparing with the hardware integrator of HXR flux. The temporal evolution of HXR flux in different energy ranges can be obtained with high time resolution by this dedicated HXR spectrometer. The response of runaway electron transport with different energy during the sawtooth activities can be studied. The energy evolution of runaway electrons during the plateau phase of runaway current can be obtained.

Fast quantitative ROS detection based on dual-color single rare-earth nanoparticle imaging reveals signaling pathway kinetics in living cells.

Reactive oxygen species (ROS), and notably hydrogen peroxide H2O2, are cellular second messengers that are known to control a variety of signaling processes. They can finely regulate the dynamics of signal transduction, cell response and ultimately tissue function. However, there are very few local, quantitative and time-resolved descriptions of their cellular organization at the scale of molecular reactions, due to the lack of efficient sensors. We thus developed a novel nanoprobe-based ROS detection system using the simultaneous imaging of single lanthanide nanoparticles (YAG:Ce and chemically reduced Gd0.6Eu0.4VO4). We reveal that both particle luminescence signals are controlled by their H2O2 local environment. By simultaneously tracking their luminescence, we devised a new approach providing a quantitative (0.5 μM accuracy in the 1-10 μM range) H2O2 measurement with a 500 ms time resolution, surpassing all existing methods by two orders of magnitude, and revealing previously inaccessible molecular events controlling ROS concentration. We used this nanoprobe in living cells to track fast signaling pathways, by measuring the dynamics of H2O2 intracellular concentrations, induced by endothelin-1 (ET-1) stimulation. We thus revealed the mechanisms controlling ROS production, notably the activity modulation of the ROS-producing enzyme NADPH oxidase by fast (<10 s) EGFR transactivation, and measured quantitatively their kinetic parameters through a minimal analytical model. Altogether, these results illustrate how lanthanide nanoparticle-based sensors are a powerful tool to dynamically probe molecular mechanisms shaping the oxidative cell response.

V4: The Small Angle Scattering Instrument (SANS) at BER II

V4 is a small-angle neutron scatting instrument with an accessible range of scattering vector 0.01 nm-1 &lt; Q &lt; 8.5 nm-1. Outstanding features of the instrument are the polarized neutron option and the list mode data acquisition, allowing for time-resolved measurements with µs time resolution.

A single-pulse shock tube coupled with high-repetition-rate time-of-flight mass spectrometry and gas chromatography for high-temperature gas-phase kinetics studies.

Shock tubes are frequently used to investigate the kinetics of chemical reactions in the gas phase at high temperatures. Conventionally, two complementary arrangements are used where either time-resolved intermediate species measurements are conducted after the initiation of the reaction or where the product composition is determined after rapid initiation and quenching of the reaction through gas-dynamic processes. This paper presents a facility that combines both approaches to determine comprehensive information. A single-pulse shock tube is combined with high-sensitivity gas chromatography/mass spectrometry for product composition and concentration measurement as well as high-repetition-rate time-of-flight mass spectrometry for time-dependent intermediate concentration determination with 10 μs time resolution. Both methods can be applied simultaneously. The arrangement is validated with investigations of the well-documented thermal unimolecular decomposition of cyclohexene towards ethylene and 1,3-butadiene at temperatures between 1000 and 1500 K and pressures ranging from 0.8 to 2.4 bars. The comparison shows that the experimental results for both detections are in very good agreement with each other and with literature data.

Optimization and application of cooled avalanche photodiodes for spectroscopic fluctuation measurements with ultra-fast charge exchange recombination spectroscopy.

The Ultra-Fast Charge Exchange Recombination Spectroscopy (UF-CHERS) diagnostic is a highly specialized spectroscopic instrument with 2 spatial channels consisting of 8 spectral channels each and a resolution of ∼0.25 nm deployed at DIII-D to measure turbulent ion temperature fluctuations. Charge exchange emissions are obtained between 528 and 530 nm with 1 μs time resolution to study plasma instabilities. A primary challenge of extracting fluctuation measurements from raw UF-CHERS signals is photon and electronic noise. In order to reduce dark current, the Avalanche Photodiode (APD) detectors are thermo-electrically cooled. State-of-the-art components are used for the signal amplifiers and conditioners to minimize electronic noise. Due to the low incident photon power (≤1 nW), APDs with a gain of up to 300 are used to optimize the signal to noise ratio. Maximizing the APDs' gain while minimizing the excess noise factor (ENF) is essential since the total noise of the diagnostic sets a floor for the minimum level of detectable broadband fluctuations. The APDs' gain should be high enough that photon noise dominates electronic noise, but not excessive so that the ENF overwhelms plasma fluctuations. A new generation of cooled APDs and optimized preamplifiers exhibits significantly enhanced signal-to-noise compared to a previous generation. Experiments at DIII-D have allowed for characterization and optimization of the ENF vs. gain. A gain of ∼100 at 1700 V is found to be near optimal for most plasma conditions. Ion temperature and toroidal velocity fluctuations due to the edge harmonic oscillation in quiescent H-mode plasmas are presented to demonstrate UF-CHERS' capabilities.

Watching Paint Dry: The Impact of Diiodooctane on the Kinetics of Aggregate Formation in Thin Films of Poly(3-hexylthiophene)

We have investigated how the addition of 1,8-diiodooctane (DIO) alters the formation of disordered and ordered phases in a film of poly(3-hexyl-thiophene-2,5-diyl) (P3HT). By combining in situ time-resolved absorption spectroscopy with 60 ms time resolution, optical and transmission electron microscopy and spatially resolved photoluminescence spectroscopy, we show that, in addition to the excitonic coupling, the film formation process during spin-coating as well as the subsequent long-time film drying process differ significantly when DIO is added to a solution of P3HT. During spin-coating, the addition of DIO reduces the actual time for transformation from disordered to ordered phase, even though it increases the time until the disorder–order transition sets in. In place of a solidification front, we observe an all-over solidification throughout the entire film. The phase separation between nonaggregated and aggregated phase increases when using DIO, with compositional variation in the content of aggrega...

Measurement of duration and signal‐to‐noise ratio of astronomical transients using a Spectral Kurtosis spectrometer

AbstractFollowing our prior theoretical and instrumental work addressing the problem of automatic real‐time radio frequency interference (RFI) detection and excision from astronomical signals, the wideband Spectral Kurtosis (SK) spectrometer design we proposed is currently being considered as an alternative to the traditional spectrometers when building the new generation of radio instruments. The unique characteristic of an SK spectrometer is that it accumulates both power and power‐squared, which are then used to compute an SK statistical estimator proven to be very effective in detecting and excising certain types of RFI signals. In this paper we introduce a novel measurement technique that exploits the power and power square statistics of an SK spectrometer to determine durations and signal‐to‐noise ratios of transient signals, whether they are RFI or natural signals, even when they are below the time resolution of the instrument. We demonstrate this novel experimental technique by analyzing a segment of data recorded by the Expanded Owens Valley Solar Array Subsystem Testbed (EST) during a solar radio burst in which microwave spike bursts occurred with durations shorter than the 20 ms time resolution of the instrument. The duration of one well‐observed spike is quantitatively shown to be within a few percent of 8 ms despite the 20 ms resolution of the data.

Investigation of driving forces of cerebrospinal fluid motion by power and frequency mapping based on asynchronous phase contrast technique.

To classify the cardiac- and respiratory-driven cerebrospinal fluid (CSF) motions, asynchronous 2D phase contrast (PC) of magnetic resonance imaging (MRI) with 217 ms time resolution in conjunction with power and frequency mapping was performed for 7 healthy subjects under respiration guidance. In the frequency domain, the cardiac-driven motion was at around 1.29±0.21 Hz and respiratory-driven motion was at 0.16±0.01 Hz under 6 sec respiratory cycle. Two different techniques were proposed for characterizing the motions; one was power-map (P-map) depicting integrated power spectrum in a selected band, and the other was frequency-map (F-map) delineating the frequency of maximum peak in power spectral density (PSD). These maps visualized the anatomical distributions of the two motions. Portions of the cardiac- and respiratory-driven CSF motions in the spinal subarachnoid space were 58.1±22.2 and 9.50±3.83 %, respectively. Power and frequency mapping clearly classified the cardiac-driven and respiratory-driven CSF motions.

Mechanical Response of Shape Memory Alloys Under a Rapid Heating Pulse - Part II

We report on a new experimental system for exploring the thermo-mechanical response of shape memory wires under a rapid heating pulse. The new setup enables the force and displacement generated by the phase transforming wire to be measured with a μs time resolution. In addition, part of the tests incorporated high-speed infrared and visible light photography. The experimental system enables exploring several unique dynamic effects. In particular, stress levels of 1.6 GPa with negligible plastic deformation and elastic strain rates of 103 s−1 are observed. Force measurements reveal a dead time of ~20 μs between the end of the electric pulse and the onset of the stress rise in the SMA wire. The high strain rate, generated by the phase transformation, induces string-like vibrations of the SMA wire that result in strong stress vibrations. The developed experimental system opens the way for studying the kinetics of the phase transformation.

Single Particle Plasmon Sensors as Label-Free Technique To Monitor MinDE Protein Wave Propagation on Membranes.

We use individual gold nanorods as pointlike detectors for the intrinsic dynamics of an oscillating biological system. We chose the pattern forming MinDE protein system from Escherichia coli (E. coli), a prominent example for self-organized chemical oscillations of membrane-associated proteins that are involved in the bacterial cell division process. Similar to surface plasmon resonance (SPR), the gold nanorods report changes in their protein surface coverage without the need for fluorescence labeling, a technique we refer to as NanoSPR. Comparing the dynamics for fluorescence labeled and unlabeled proteins, we find a reduction of the oscillation period by about 20%. The absence of photobleaching allows us to investigate Min proteins attaching and detaching from lipid coated gold nanorods with an unprecedented bandwidth of 100 ms time resolution and 1 h observation time. The long observation reveals small changes of the oscillation period over time. Averaging many cycles yields the precise wave profile that exhibits the four phases suggested in previous reports. Unexpected from previous fluorescence-based studies, we found an immobile static protein layer not dissociating during the oscillation cycle. Hence, NanoSPR is an attractive label-free real-time technique for the local investigation of molecular dynamics with high observation bandwidth. It gives access to systems, which cannot be fluorescently labeled, and resolves local dynamics that would average out over the sensor area used in conventional SPR.

Collision dynamics of laser produced carbon plasma plumes

We present preliminary experimental observations of the collision processes between two orthogonal laser produced plasmas in a low pressure neutral gas background. A Nd:YAG laser, 340 mJ, 3.5 ns, at 1.06 μm, operating at 10 Hz, is used in the experiments. The main laser beam is divided in two beams by a 50% beam splitter, and then focused over two rotating graphite targets, with characteristic fluence 3.5 J/cm2. Experiments are conducted in a range from a base pressure of 0.3 mTorr, up to 50 mTorr argon. The dynamics of the laser plasmas is characterized by time resolved and time integrated optical emission spectroscopy (OES), with 20 ns and 10 ms time resolution, and 50 ns time resolved plasma imaging of visible plasma emission. Clear effects of the neutral gas background on the postcollision plasma dynamics are identified. The overall dynamics of the post-collision plasma is found to be consistent with high collisionality of the carbon plasma plumes, which results in full stagnation on collisioning.

Variations in Fusion Pore Formation in Cholesterol-Treated Platelets

Exocytosis is a highly regulated intercellular communication process involving various membrane proteins, lipids, and cytoskeleton restructuring. These components help control granule fusion with the cell membrane, creating a pore through which granular contents are released into the extracellular environment. Platelets are an ideal model system for studying exocytosis due to their lack of a nucleus, resulting in decreased membrane regulation in response to cellular changes. In addition, platelets contain fewer granules than most other exocytosing cells, allowing straightforward measurement of individual granule release with carbon-fiber microelectrode amperometry. This technique monitors the concentration of serotonin, an electroactive molecule found in the dense-body granules of platelets, released as a function of time, with 50 μs time resolution, revealing biophysical characteristics of the fundamental exocytotic process. Variations in fusion pore formation and closure cause deviations from the classic current versus time spike profile and may influence diffusion of serotonin molecules from the site of granule fusion. Physiologically, the delivery of smaller packets of chemical messengers or the prolonged delivery of chemical messengers may represent how cells/organisms tune biological response. The goals of this work are twofold: 1) to categorize secretion features that deviate from the traditional mode of secretion and 2) to examine how changing the cholesterol composition of the platelet membrane results in changes in the pore formation process. Results herein indicate that the expected traditional mode of release is actually in the minority of granule content release events. In addition, results indicate that as the cholesterol content of the plasma membrane is increased, pore opening is less continuous.