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A pulse generator simulating slit-scan chromosome analysis signals.

A simple circuit is described for generating a variety of electronic pulses to test hardware and software for slit-scan chromosome analysis in a flow cytometer. The pulse shape can be changed to have different numbers of local minima, thereby simulating fluorescence pulses from acrocentric, monocentric, and dicentric chromosomes. Long pulses simulate aggregates of chromosomes. The pulse repetition rate as well as the pulse amplitude is variable. Although the circuitry is built with only three integrated circuits, the pulse-to-pulse variation in shape and height is quite small. After digitization of the analog signals, the constructed histograms of pulse integrals show a relative coefficient of variation below 1%. This signal generator provides a valuable tool for a number of electronic test applications that would otherwise require expensive standard particles analyzed in a well-tuned flow cytometer.

Animals↗

Pulse modulation of the excitation light source boosts the sensitivity of an arc lamp-based flow cytometer.

It has been found that the type of short arc high-pressure lamps used in some flow cytometers can be pulsed to reach intensities many times higher than that measured when they are operated on constant power. A 75 W mercury-xenon lamp was fed 20 microseconds current pulses super-imposed on its rated current of 5.4 A. Pulses of 50 A produced a 75-fold increase of the emission intensity at the excitation wavelength of FITC, which means that the excitation intensity in the flow chamber at this wavelength exceeded 100 mW. At the major emission lines of mercury, i.e., 366, 436, and 546 nm, the increase was about 25-fold, corresponding to intensities of the order of 300 mW. The light pulses were found to be reproducible to within 2% and the intensity was independent of the pulse frequency up to at least 250 pulses/s. Operated in this mode the lamp produced more than 1 x 10(8) 30 A pulses, which is sufficient to measure some 10,000 typical size samples. The rise time of the light pulses was about 5 microseconds. This was sufficiently short that the leading edge of the light scattering signal from a cell entering the excitation focus of the flow cytometer could be used to trigger the current pulse so that the cells were exposed to the full light pulse intensity while they were still within the focus.(ABSTRACT TRUNCATED AT 250 WORDS)

Flow Cytometry↗

Flow cytometric measurement of NK cell immunoconjugates by pulse width processing.

Pulse width analysis, in flow cytometry, has been widely used for optimal cell size resolution in cell kinetics analysis. Pulses, generated by scattered light or fluorescence of cells, are electronically analyzed for their height and width. The information generated from these two properties of the pulses is utilized to distinguish signals from single cells vs. signals from cell clumps or aggregates. Pulse width, unlike pulse height, is more sensitive to differences in cell diameter, and therefore can discriminate very small differences in it, which pulse height cannot. We have exploited this property of pulse widths to measure immunoconjugates between NK cells and their targets. Discrimination of the free target cells from the conjugated ones is possible by the pulse widths of only light scatter signals, both forward and/or orthogonal. This resolution was not obtained if pulse height of the same signals was visualized. Using this resolution it was possible to distinguish single cells from the aggregates between target and effector cells. We propose that this is a better method for distinguishing conjugates than the method in which prior vital staining of cells is used.

Animals↗

Continuous wave (CW) and pulsed laser effects on vascular tissues and occlusive disease in vitro.

Human arterial segments with occlusive defects and acute dog hearts were exposed, in vitro, to high-energy pulsed and continuous wave (CW) laser beams at argon (514 nm) and Nd-YAG (1,064 nm) wavelengths, using various pulse powers, durations and pulse repetition rates. The laser effects included vaporization of plaques in the arterial segments and penetration of the pericardial sac, evaporation of pericardial fluid, and discoloration of tissue with crater-like lesions in the impact zone, all as a result of vaporization of heart muscle tissues. The areas affected and depth of penetration depended on the wavelength, power, pulse duration, and mode of energy deposition. Focused nanosecond Nd-YAG laser pulses at repetition rates of 40-50 Hz caused ablation or vaporization of hard plaques and kidney stones in air and saline. Picosecond (mode-locked) argon laser pulses at repetition rates of 3.8 MHz--average power 6.5 W, peak power of 230 W--caused effective vaporization of hard plaques and kidney stones in air and saline. Picosecond argon laser pulses--average power 1 W, peak power 250 W--were not effective in vaporization. Transmission characteristics of the various types of laser pulses through fiber optic waveguides were determined. The energy and power density required to vaporize fatty and hard plaques and kidney stones were tabulated as a function of laser wavelength, pulse energy, duration, and repetition rates.

Animals↗

ED50 study of femtosecond terawatt laser pulses on porcine skin.

BACKGROUND AND OBJECTIVES: Terawatt (TW) lasers have become commonplace since the development of the chirped-pulse amplification method using Ti:sapphire and Nd:glass laser rods. We have measured the minimum visible lesion (MVL) thresholds for porcine1The animals involved in this study were procured, maintained, and used in accordance with the Federal Animal Welfare Act and the "Guide for the Care and Use of Laboratory Animals" prepared by the Institute of Laboratory Animal Resources-National Research Council. Brooks City-Base, TX has been fully accredited by the Association for Assessment and Accreditation of Laboratory Animal Care, International (AAALAC) since 1967. (Yucatan mini-pig) skin using TW laser pulses. STUDY DESIGN/MATERIALS AND METHODS: Our system produced laser pulses at 810 nm and sub-50 femtoseconds. These 1-2 TW laser pulses created multiple self-focusing (SF) filaments during propagation and were directed on the flanks of mini-pigs under anesthesia. We measured the pulse energies necessary to determine the ED(50) skin damage thresholds. RESULTS: The MVL ED(50) threshold at 1 hour was 8 mJ and increased to 21 mJ after 24 hours. Histological sections were obtained after 1-hour and 24-hour readings. CONCLUSIONS: The damage patterns on the skin indicated the number of filaments in the laser pulse. Many of the pulses produced only superficial damage that disappeared in 24 hours and that nearly three times the pulse energy was required to cause subdural or cellular damage. With further research, non-thermal tissue ablation using TW laser pulses could provide a viable alternative to current techniques of laser use in dermatology.

Animals↗

Errors in the measurements of T2 using multiple-echo MRI techniques. I. Effects of radiofrequency pulse imperfections.

In principle, multiple-echo magnetic resonance imaging (MRI) can be used to estimate the spin-spin relaxation time, T2, which can then be used for quantitative tissue characterization. Although multiple echoes can be used to enhance the signal-to-noise ratio in an image, by echo addition, rf pulse imperfections modify the echo amplitudes resulting in significant errors in the estimate of T2. Imperfect 180 degree pulses do not completely invert the transverse magnetization so that the magnitude of the transverse component is reduced and a longitudinal component is generated. Successive application of such imperfect pulses generates many components that interact in a complex manner. The amplitudes of successive echoes are affected whenever the transverse components refocus, whereas the longitudinal components may be rotated into the transverse plane by subsequent pulses and may often add to the image signal or give rise to an image artifact. These effects have been analyzed theoretically and have been demonstrated for a wide range of rf pulse imperfections using both simple and composite pulses, through computer simulations based on the numerical solution of the Bloch equations. The theoretical and simulation results have been substantiated through experiments performed on a mineral oil phantom using a resistive prototype MR scanner operating at 6.35 MHz. In this paper we report the effects of improper pulse amplitude or duration for nonselective rf pulses on resonance. We separately describe the other types of imperfections caused by off-resonance effects and the use of selective pulses.

Humans↗

Effect of differential saturation on the spatial localization performance of depth pulses.

Computer simulations of Depth pulse B (theta; (2 theta [+/- x, +/- y])2; acquire) and other Depth pulses, verified by experimental surface coil NMR studies utilizing phantom samples, reveal that the spatial localization performance of Depth pulses degrades when the repetition time is short relative to T1 because of differential saturation, i.e. T1 discrimination effects. Simulations of Depth pulse A (theta; 2 theta [+/- x, +/- y]; acquire) and Depth pulse B indicate that there is no phase-cycled pulse sequence delivery order which negates the untoward effect of T1 discrimination on spatial localization performance. The results of this study demonstrate the importance of consistent magnetization preparation prior to the delivery of each phase-cycled multiple pulse sequence in a Depth pulse cycle for obtaining optimal spatial localization performance. The untoward effects of inconsistent magnetization preparation, resulting from T1 discrimination, may be ameliorated by the application of many Depth pulse cycles.

Computer Simulation↗

Spatially selective RF pulses and the effects of digitization on their performance.

Spectrometers make use of D/A converters to generate RF and gradient shapes. This paper examines by exact simulations the time and amplitude digitization effects, inherent to the use of D/A converters, on the performance of amplitude modulated (AM) frequency selective RF pulses. By making use of Fourier theory and the small tip angle approximation, an approximate model of these effects on the magnetization slice profiles is derived and verified for several pulse types by computer simulations. This approximate model will be used to derive requirements for D/A converters with respect to spatial localization. The dynamics of the spin system allows pulse width modulation (PWM) as an alternative to AM for pulse envelope encoding. The effects of PWM on the slice profile are examined and compared with conventional AM pulses. It is shown by simulation and measurement that adiabatic PWM pulses can be found. In contrast to AM modulated adiabatic pulses, adiabatic PWM pulses have side bands with the same slice quality as the main slice and might therefore be useful as multislice selective pulses.

Magnetic Resonance Spectroscopy↗

Pulse timing and optical interface between a neodymium: yttrium aluminum garnet laser and a Fourier transform ion cyclotron resonance mass spectrometer.

Laser desorption/ionization combined with pulsed (time-of-flight or Fourier transform ion cyclotron resonance) mass spectrometric detection is a powerful technique for analysis of involatile compounds and mixtures. Such experiments were originally conducted with pulsed CO2 lasers. Although a pulsed CO2 laser can be operated in single-shot mode, Nd: YAG lasers perform best with multiple flashes for warm-up before the final Q-switch output light pulse, thus creating the need to synchronize the desired final laser-output pulse with the event sequence for mass spectrometric analysis. In this paper, we describe a new and simple interface (both optical and electronic components) between a Continuum (formerly Quantel) Model YG 660A Nd:YAG laser and an Extrel FTMS-2000 mass spectrometer. The optics are modified from a prior pulsed CO2 laser interface from Extrel. Synchronization between the Nd:YAG laser and the mass spectrometer event sequence is achieved by means of a simple timing circuit that uses an inexpensive pulsing device and is triggered by pulses generated directly from the Extrel 1280 data system and cell controller, in contrast to the only prior published method that required an auxiliary microcomputer. The present interface method is highly flexible, and makes possible complex sequence events involving laser pulses for e.g.: desorption/ionization of solids; photoionization of gaseous neutrals; and photodissociation and photodetachment of gaseous ions.

Lasers↗

Use of amplitude and frequency transformations to generate adiabatic pulses of wide bandwidth and low RF power deposition.

An amplitude transformation of adiabatic pulses stated previously [Baum et al., Phys. Rev. A 32, 3435-3447 (1985)] is generalized using the Bloch equations. Both amplitude and frequency transformations are used to create new adiabatic pulses of wide bandwidth and low RF power deposition. Several adiabatic pulses (including the tanh/tan pulse used for construction of BIR-4, SSAP, and BISEP pulses) are transformed into new pulses. These new pulses are demonstrated numerically and experimentally to operate at a significantly lower RF power level while maintaining the same performance or over a wider bandwidth while using the same RF power. They are expected to be useful for in vivo NMR experiments, especially for applications involving wide frequency dispersion and pulse sequences composed of many adiabatic half-passage pulses.

Magnetic Resonance Imaging↗

Design of selective adiabatic inversion pulses using the adiabatic condition

Adiabatic RF pulses play an important role in spin inversion due to their robust behavior in the presence of inhomogeneous RF fields. These pulses are characterized by the trajectory swept by the tip of the Beff vector and the rate of motion along it. In this paper, we describe a method by which optimized modulation functions can be constructed to render insensitivity to B1 inhomogeneity over a predetermined B1 range and over a wide band of frequencies. This is accomplished by requiring that the optimized pulse fulfill the adiabatic condition over this range of B1 inhomogeneity and over the desired frequency band for the complete duration of the pulse. A trajectory similar to the well-known sech/tanh adiabatic pulse, i.e., a half-ellipse, is used. The optimization process improves the slice profile by optimizing the rate of motion along this trajectory. The optimized pulse can be tailored to the specific design requirements; in particular, the transition sharpness may be traded off against the inverted bandwidth. Two design examples, including experimental results, demonstrate the superiority of the optimized pulses over the conventional sech/tanh pulse: in the first example, a large frequency band is to be inverted using a weak RF amplitude in a short time. In the second example, a pulse with a very sharp transition is required. Copyright 1997 Academic Press. Copyright 1997Academic Press

Journal Article↗

Broadband echo sequence using a pi composite pulse for the pure NQR of a spin I = 32 powder sample

This work presents a numerical approach to optimizing sequences with composite pulses for the pure NQR of a spin I = 32 powder sample. The calculations are based on a formalism developed in a previous paper, which allows a fast powder-averaging procedure to be implemented. The framework of the Cayley-Klein matrices to describe space rotations by 2 x 2 unitary and unimodular complex matrices is used to calculate the pulse propagators. The object of such a study is to design a high-performance echo sequence composed of a single preparation pulse and a three-pulse composite transfer pulse. We mean a sequence leading to a large excitation bandwidth with a good signal-to-noise ratio, a flat excitation profile near the irradiation frequency, and a good linearity of the phase as a function of frequency offset. Such a composite echo sequence is intended to give a better excitation profile than the classical Hahn (θ)-tau-(2θ) echo sequence. It is argued that in pure NQR of a powder sample, the sequence must be optimized as a whole since both the excitation and the reception of the signal depend on the relative orientation of the crystallites with respect to the coil axis. To our knowledge, this is the first time such a global approach is presented. An extensive numerical study of the composite echo sequence described above is performed in this article. The key of the discrimination between the sequences lies in using the first five reduced moments of the excitation profile as well as an estimator of the phase linearity. Based on such information, we suggest that the echo sequence that best fulfills our criterion is (1)(0)-tau-(0.35)(0)(2.1)(pi)(0.35)(0), the pulse angles omega(RF)t(p) being in radians. The subscripts are the relative pulse phases. We outlined the way to implement the spin echo mapping method to reconstruct large spectra with this sequence, and it is shown that it reduces the acquisition time by a factor of 1.7 if compared to the classical Hahn echo. Some other broadband echo sequences are also briefly discussed. We also study the effect of a small delay between the pulses of the composite pulse to take into account the experimental constraint imposed by the spectrometer. To be complete, we give an estimation of the performances of the Hahn echo, the stimulated echo, and the composite echo sequence optimized for a powder sample when applied to a monocrystal. Experiments performed on chloranil at ambient temperature confirm the predicted excitation profiles. Copyright 2000 Academic Press.

Journal Article↗

In vitro effects of high-energy pulsed ultrasound on human squamous cell carcinoma cells.

Human squamous cell carcinoma cells cloned from the hypopharynx (FaDu) and oral cavity (SCC-4) were exposed to high-energy pulsed ultrasound (HEPUS) in vitro to evaluate the effects of various physical parameters on cell viability. Such included the number of pulses, voltage applied, pulse repetition rate and cell density. The experimental piezoelectric ultrasound transducer used in the experiments generated pulses with a high negative pressure amplitude. By varying the repetition frequency from 0.6 to 8 Hz, cell viability was found to be least when pulse repetition was approximately 1 Hz. An increase in transducer voltage resulted in a linear decrease in cell viability. The cell survival rate dropped exponentially as a function of the number of pulses applied, reaching 4.2% after 2000 pulses. The cell survival rate exhibited no significant dependence on cell density when cells ranged from 1 to 3.5 x 10(6) cells ml-1. Data obtained with trypan blue dye exclusion were confirmed by measurements of intracellular lactate dehydrogenase released into an extracellular fluid supernatant. By applying HEPUS to tumor cells, almost complete destruction of the cells could be achieved in vitro. The degree of cell destruction achieved depended significantly on the number of pulses administered, the pulse repetition rate and the transducer voltage applied.

Carcinoma, Squamous Cell↗

A short red light pulse during dark phase of LD-cycle perturbs the hamster's circadian clock.

In this study we investigated the influence of red light< which naturally occurs during dawn and dusk, on locomotor activity and body temperature rhythms of Djungarian hamsters (Phodopus sungarus). A single weak red light pulse given 2 h before regular lights on had acute as well as long-term effects persisting for several days following exposure. The hamsters immediately stopped their locomotor activity, accompanied by a drop in body temperature. In the following undisturbed nights (ID 16:8) the nocturnal activity stopped earlier than usual. This lasting effect of the light pulse was more pronounced than the acute effect. The activity phase compressed gradually during 3 to 5 days after the light pulse was administered while time of activity onset was almost unaffected. It took 6 to 11 days for complete recovery of the original activity phase. The maximal activity compression and the recovery period depended on the duration of the single red light pulse and its intensity. Red light pulses of 15 min duration were about twice effective as 1 min pulses; and the effect of a red light pulse of 130 mW/m2 was about 1.5 times stronger than a 30 mW/m2 red light pulse. The maximal value of activity phase compression reached in this experiment was 2.5 + 0.2 h with a recovery period of 11.1 +/- 0.3 days following a given red light pulse of 90 mW/m2 and 15 min. The "morning oscillator" seems to be persistently affected. This indicates a very high photosensitivity of the Djungarian hamster's circadian system to red light.

Animals↗

Large phase-shifts of circadian rhythms caused by induced running in a re-entrainment paradigm: the role of pulse duration and light.

Bouts of induced wheel-running, 3 h long, accelerate the rate of re-entrainment of hamsters' activity rhythms to light-dark (LD) cycles that have been phase-advanced by 8 h (Mrosovsky and Salmon 1987). The bouts of running are given early in the first night of the new LD cycle, and by the second night the phase advance in activity onset already averages 7 h. Such large shifts contrast with the mean phase advance of less than 1 h at the peak of the phase response curve when hamsters in constant darkness (DD) experience 2-h pulses of induced activity (Reebs and Mrosovsky 1989). The present paper investigates pulse duration and light as possible causes for the discrepancy in shift amplitude between these two studies. In a first experiment, pulses of induced wheel-running 1 h, 3 h, or 5 h long were given at circadian times (CT) 6 and 22-2 to hamsters free-running in DD. Pulses given at CT 6 caused phase-advances of up to 2.8 h, whereas pulses at CT 22-2 resulted in delays of up to 1.0 h. Shifts after 3-h and 5-h pulses did not differ, but were larger than after 1-h pulses, and larger than after the 2-h pulses given in DD by Reebs and Mrosovsky (1989). Thus 3 h appears to be the minimum pulse duration necessary to obtain maximum phase-shifting effects. In a second experiment, the re-entrainment design of Mrosovsky and Salmon (1987) was repeated with the light portion of the shifted LD cycle eliminated.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Comparison of rectangular and exponential current pulses for evoking sensation.

There exists a paucity of quantitative information comparing the stimulating currents for different waveforms. Thus, the objectives of this study was to compare the threshold peak current (I) for sensation using rectangular and exponential; that is, capacitor-discharge, cathodal pulses of equivalent pulse duration (d). In 10 human subjects, stimuli were applied to the skin of the forearm, and I was determined alternately for each current waveform at each of several pulse durations (d). Strength-duration curves for sensation were obtained using d of 0.01-50 ms. The threshold peak current (Ir) for a rectangular pulse of duration d was compared to the threshold peak current (Icd) for a capacitor-discharge pulse of duration d, where d was the time constant; that is, the time required for the current to decrease to 1/e, or 37% of its peak value. Chronaxie, the pulse duration at which I is twice the infinite-duration current asymptote (i.e., the rheobase), was calculated for each waveform and subject using the Weiss-Lapicque expression for excitability. Icd was found to be always higher than Ir of equivalent duration. Chronaxie for the capacitor-discharge pulse was, on the average, twice that for the rectangular pulse (p less than 0.01). Moreover, the ratio Icd/Ir increased with decreasing d. These results indicate that these two waveforms are not equivalent on the basis of an equal-charge requirement for excitation, particularly at the short pulse durations. Furthermore, they suggest the need of a better expression to describe the excitability characteristics of tissues.

Adult↗

Laser-induced ablation of a steel sample in different ambient gases by use of collinear multiple laser pulses.

The sensitivity of laser-induced breakdown spectroscopy of solid samples depends on the number of ablated and excited analytes. Laser ablation of solid samples can be enhanced by using collinear multiple laser pulses, for example double or triple pulses, rather than single laser pulses with the same total laser pulse energy. The ablation rates and the plasma conditions are affected by the ambient gas. In this study laser ablation was examined by varying the interpulse separation of the multiple pulses, within double and triple-pulse bursts, and the gas mass density at constant gas pressure. Different ambient gases and gas mixtures consisting of argon, oxygen, and nitrogen were used to study their effect on ablation rates. In a pure argon atmosphere (99.999% v/v Ar) the ablation burst number required to penetrate a steel plate of thickness 100 microm is reduced by a factor of approximately six by use of triple-pulse bursts with a symmetric interpulse separation of 15 micros rather than single pulses with the same total burst energy of 105 mJ. For double and single pulses the factors are 1.6 for Ar and 2.8 for synthetic air. Analyte lines are 4 to 8 times more intense if an argon atmosphere, rather than air, is used.

Journal Article↗

LIBS using dual- and ultra-short laser pulses.

Pre-ablation dual-pulse LIBS enhancement data for copper, brass and steel using ns laser excitation are reported. Although large enhancements are observed for all samples, the magnitude of the enhancement is matrix dependent. Whereas all of the dual-pulse studies used ns laser excitation we see interesting effects when using ps and fs laser excitation for single-pulse LIBS. LIBS spectra of copper using 1.3 ps and 140 fs laser pulses show much lower background signals compared to ns pulse excitation. Also, the atomic emission decays much more rapidly with time. Because of relatively low backgrounds when using ps and fs pulses, non-gated detection of LIBS is shown to be very effective. The plasma dissipates quickly enough using ps and fs laser pulses, that high pulse rates, up to 1,000 Hz, are effective for increasing the LIBS signal, for a given measurement time. Finally, a simple near-collinear dual-pulse fiber-optic LIBS probe is shown to be useful for enhanced LIBS measurements.

Journal Article↗