Laser flow cytometric light scatter and fluorescence pulse width and pulse rise-time sizing of mammalian cells.

In laser flow cytometry, an increasingly popular technique of analytical cytology, quantitative measurements of interest include cell and nuclear diameters. Electronic circuitry for a new cell sizing technique has been developed which measured the time that signal pulses from either fluorescence or light scatter sensors exceed a preset constant fraction of the peak signal amplitude (pulse width) or the time that it takes a signal to rise between constant fractions of the peak signal amplitude on the rising side of the pulse (pulse rise-time). These pulse width or pulse rise-time measurements were related to cell or nuclear diameters and were used in combination to determine nuclear size to cell size ratios. This method of sizing was found to be independent of fluorescent or light-absorbing stain intensity, linearly related to cell or nuclear diameter, and capable of resolving small diameter differences.     

Analysis of Wrist Pulse Signal: Emotions and Physical Pain

BackgroundPulse diagnosis (wrist pulse signal) is a well-known traditional technique used for a health examination. It has the potential to detect cardiac and non-cardiac diseases.ObjectiveA study was conducted to investigate human emotions using wrist pulse signal assessment. The aim was to categorize anxiety, boredom, physical pain, and reference state by processing and analysis of acquired signals.MethodA protocol was designed to induce emotions. Data were acquired from 24 healthy volunteers. Signals were processed and further analyzed using paired t-test and Analysis of Variance (ANOVA). Machine learning algorithms, Linear Discriminant Function (LDF), Quadratic Discriminant Function (QDF), and Support Vector Machine using kernel Gaussian radial basis function (RBF-SVM) were used to evaluate significant features and classify the emotions.ResultsComputing significant plus ranked features performed better over randomly selected features for pairwise emotion classification. Here, the QDF classifier outperforms LDF. Additionally, ANOVA validated the effectiveness of statistically prominent features to classify emotional states. Ratio_Pulse_Strength, total_power, Spectral Entropy, and meancd5 came out as the four most significant features to classify the emotion “Anxiety”, “Boredom”, “Pain”, and “Reference” with positive prediction rate of 100%, 73%, 100%, and 86% respectively using RBF-SVM in the user-independent model.ConclusionPreviously, WPS has been used mainly to detect physical abnormality in the human body. The results endorse the potential of user-independent human emotion detection using a wrist pulse signal. The present work was focusing on a few emotional states. Results are encouraging and may be well applied to many more states.     

High quality photoplethysmograph signals from a laser Doppler flowmeter: preliminary studies of two simultaneous outputs from the finger

An improvement in the quality of a photoplethysmograph signal derived from a laser Doppler flowmeter probe has been achieved by incorporating an auxiliary fibre in the probe head. This fibre is positioned at an optimum distance from the laser light transmitting fibre and overcomes the problem of high frequency signals which mask the detailed features of the photoplethysmograph pulse when a small fibre separation is used. These two signals may be recorded simultaneously from the same site and a correlation between the Doppler output and the photoplethysmograph amplitude has been demonstrated in the finger. The amplitude is shown to be affected by the relative position of the point of measurement with respect to the heart, a factor which does not appear to influence significantly the Doppler output.     

Characteristics and mechanisms of the two types of photoelectric differential response of bacteriorhodopsin-based photocell

Bacteriorhodopsin (BR)-based photocells have been assigned possessing differential photoelectric response. But recently we found that the differential response described before, which occurred in milliseconds to seconds, outputting a positive pulse when light was on and a negative pulse when light was off, was not the intrinsic property of the BR molecule. It was partially caused by the measuring circuit. By measuring the photoelectric response signal of the BR film photocell to a short laser pulse, the impulse response function of the BR film photocell was obtained by data fitting with MATLAB software. A simulation system was accordingly developed. The output response signals of the BR film photocell under different stepping incident light were calculated. By simulation and analysis, it was concluded that the differential response caused by the intrinsic property of the BR molecule happened in microseconds time scale, and it produced a negative pulse when light was on and a positive pulse when light was off. It was much faster but much weaker than that described before.     

Threshold assessment,categorical perception,and the evolution of reliable signaling

Animals often use assessment signals to communicate information about their quality to a variety of receivers, including potential mates, competitors, and predators. But what maintains reliable signaling and prevents signalers from signaling a better quality than they actually have? Previous work has shown that reliable signaling can be maintained if signalers pay fitness costs for signaling at different intensities and these costs are greater for lower quality individuals than higher quality ones. Models supporting this idea typically assume that continuous variation in signal intensity is perceived as such by receivers. In many organisms, however, receivers have threshold responses to signals, in which they respond to a signal if it is above a threshold value and do not respond if the signal is below the threshold value. Here, we use both analytical and individual-based models to investigate how such threshold responses affect the reliability of assessment signals. We show that reliable signaling systems can break down when receivers have an invariant threshold response, but reliable signaling can be rescued if there is variation among receivers in the location of their threshold boundary. Our models provide an important step toward understanding signal evolution when receivers have threshold responses to continuous signal variation.     

The variability of mating signals and preferences in the brown planthopper,Nilaparvata lugens (Homoptera: Delphacidae)

Male and female brown planthoppers, Nilaparvata lugens(Stål) (Homoptera: Delphacidae), exchange substrate-transmitted signals prior to mating. The pulse repetition frequency of the male song is known to be involved in mate recognition and also to vary among geographical populations. Here the variability of male signals, female signals, and female preferences has been examined within a population. Female preference variation has been partitioned into variation in mean preference and variation in the window of preference of individuals. The genetic component of variation has been examined using isofemale lines. Male signal variation was limited (CV=8%) and was mainly within individuals. Female signal variation was greater (CV=15%). Female mean preference varied little (CV=10%) and was closely matched to the male signal mean, but the preference window was wide (> 4 male signal standard deviations on average) and variable (CV=56%). There was evidence for genetic variation only for preference window. These results are discussed in relation to theories of signal system evolution.     

Physiological MR signal variations within the brain at 3 T.

The echoplanar technique in magnetic resonance (MR) imaging allows the acquisition of a series of images from a selected slice with a temporal resolution of 10/s. Simultaneous recording of physiological information on pulse and respiration allows correlation of the MR signal intensity with physiological signals, which can be obtained for each pixel examined. Such correlations can be found within the cerebrospinal fluid (CSF) spaces and within vessels if a flow-sensitive MR measurement technique is used. The use of an MR scanner with a field strength of 3 T improves the signal/noise ratio, but there is a stronger signal decay due to local magnetic inhomogeneities. This study shows that 3-T systems can be used for correlation of MR and physiological signals and that clear differentiation between signals from CSF and from vessels can be obtained due to their strongly different signal decays.     

The use of backscattered electrons in analytical electron microscopy for the measurement of the mass of individual rat blood platelets

The backscattered electron signal, generated in individual cells, has been used to measure the dry mass of these cells. Absolute mass values were obtained by comparing the backscattered electron signals of cells to the signals of polystyrene-latex spheres of known mass. The technique was carried out in an automated analytical scanning transmission electron microscope and applied to rat blood platelets. The resulting mass distributions agreed well with the distribution measured with a method that uses the transmitted electron signal by means of densitometric analysis of electrographs. Also the range of masses was in agreement with values deduced from data in the literature. The fully automated technique has the advantage that it is direct, fast, and that thicker specimens can be measured than is possible using the transmitted electron signal. The method is intended for use in combination with quantitative electron probe X-ray microanalysis and is then able to produce elemental mass fractions of biological specimens at the subcellular level.     

The use of backscattered electrons in analytical electron microscopy for the measurement of the mass of individual rat blood platelets

Summary The backscattered electron signal, generated in individual cells, has been used to measure the dry mass of these cells. Absolute mass values were obtained by comparing the backscattered electron signals of cells to the signals of polystyrene-latex spheres of known mass. The technique was carried out in an automated analytical scanning transmission electron microscope and applied to rat blood platelets.The resulting mass distributions agreed well with the distribution measured with a method that uses the transmitted electron signal by means of densitometric analysis of electrographs. Also the range of masses was in agreement with values deduced from data in the literature.The fully automated technique has the advantage that it is direct, fast, and that thicker specimens can be measured than is possible using the transmitted electron signal. The method is intended for use in combination with quantitative electron probe X-ray microanalysis and is then able to produce elemental mass fractions of biological specimens at the subcellular level.In honour of Prof. van Duijn     

Viable sorting of intact multicellular spheroids by flow cytometry

A flow cytometric method has been developed for sorting viable, intact multicellular spheroids in order to obtain uniformly-sized populations with diameters in the range of 50-100 microns. A FACS II instrument was modified for this purpose by installing a 200-microns-diameter exit orifice and by making adjustments in the sheath flow, oscillator frequency, and number of droplets sorted. Polystyrene microspheres (44 and 88 microns diameter) and 41-96-microns-diameter spheroids could be sorted and recovered with 70-100% efficiency, an improvement over previous reports. Unstained, viable spheroids were simultaneously analyzed for small-angle forward light scatter, 90 degree light scatter, and autofluorescence using a 488-nm laser operating at 100 mW. Analysis of the data demonstrated a considerable variation in both the 90 degrees light scatter and the autofluorescence signals for a given forward angle light scattering signal. By setting narrow sort windows on the forward angle light scattering signal and either the 90 degree light scatter or autofluorescence signals, uniformly spherical spheroid populations could be recovered. These sorted populations had coefficients of variation of the mean diameter in the range of 5-9%. This represents a variation of less than one cell diameter, and is a major improvement over any other technique. There was no significant difference in the subsequent growth rates of sorted spheroids compared to the unsorted spheroids. This technique will apply when uniform populations of small spheroids are required, such as investigations of the contact effect or in the initiation of growth curve studies.     

Vibrational alarm communication in the damp-wood termite Zootermopsis nevadensis

Abstract. Vibrational alarm communication was studied in the New World, damp-wood termite Zootermopsis nevadensis (Isoptera: Termopsidae). Workers and soldiers react to disturbance such as sudden bright light or air currents by drumming their heads against the substratum. This drumming has been described as alarm signalling; its functional significance and perception by the nest mates, however, remained unclear. In the present study we analysed spectral and temporal properties and absolute amplitudes of the vibrational signals and used behavioural responses of the termites to determine the thresholds of the sense of vibration and to find out if and how the termites discriminate the conspecific alarm signals from the background noise.
The drumming signals are trains of pulses of vibrations of the substratum with a pulse repetition rate of about 20 Hz. The carrier frequency depends on the substratum; in the nests studied it was in the range 1–3 kHz. The highest vibrational amplitudes measured close to the signal emitters are usually about 10m/s² (acceleration, RMS). The threshold of the behavioural response is about 1m/s² over a wide range of frequencies (10 Hz to 5 kHz), indicating that the termites can detect these signals as vibrations of the substratum. The animals respond preferentially to temporal patterns similar those of the natural signals; temporal rather than spectral cues seem to be used for signal discrimination.     

Determination of the light pathlength elongation in leaves by measuring P700 quantitatively

P700 absorption change signals were measured at 800 nm for plant leaves and plant leaf extracts by illuminating with far-red light. The ratio of the signal amplitudes for the two sample types shows the degree of optical signal intensification. The intensification is due to optical path-length elongation, itself caused by scattering. The intensification was found to be between 6 and 15 for different leaves. For spinach leaves, the intensification varied less than 10% among samples. By replacing the air in the spongy layer of a leaf with an isotonic solution it was shown that nearly half of the intensification is due to the scattering at air-tissue interfaces. The comparison of the P700 signals of a leaf with its extract would seem to be a new technique for determining the optical scattering effects of leaves.     

Hyper-Rayleigh scattering of protein-modified gold nanoparticles

The nonlinear optical properties of protein-modified gold nanoparticles has been studied by the hyper-Rayleigh scattering (HRS) technique. HRS signals from the nanoparticles coated with goat-anti-human IgG have been obtained when pumped with a laser pulse with a wavelength of 1064 nm. The HRS signals of gold nanoparticles with IgG were larger than those of bare gold nanoparticles. This can be explained by a noncentrosymmetric effect. It was also found that the HRS signals from the IgG-coated gold nanoparticles could be greatly increased when the antigen was added due to gold nanoparticle aggregation. Our experiment found that the HRS method could produce a measurable signal with 10 microg/ml antigen added, while the colorimetric method using UV spectrum detection required 100 microg/ml of added antigen. The results show that the HRS measurement of immunogold nanoparticles could become a potential immunoassay in determining small levels of antigen in aqueous samples.     

Body slap: An innovative aggressive display by breeding male gray seals (Halichoerus grypus)

Aggression in male gray seals has been extensively studied; however it is often simplistically assumed that threat signals are mainly cephalic in nature for this species. We report on an undescribed and apparently new kind of threat signal used by male gray seals we term a Body Slap. The behavior has been observed at breeding sites in eastern England since 1993 but has not been studied ethologically or reported elsewhere. The aims of this study were to describe the behavior, test the influence of topographic variation on its frequency of occurrence, examine if it is used to signal dominance or submission, and to place it in intra‐ and interspecific contexts. Our results show Body Slaps were performed in 66.3% of interactions and by 57.2% of males; it was not performed by females. The Body Slap was positively associated with the Approach and Open‐Mouth Threat behaviors but was not related to dominance; nevertheless, display rates were greater for subsequent winners. These findings suggest that the Body Slap carries information about male resource holding potential and does not signal submission. This study furthers our understanding of geographic variants of male threat behaviors and of pinniped nonvocal communication.     

Attachment of oligonucleotide probes to poly carbodiimide-coated glass for microarray applications

Oligonucleotide-based DNA microarrays are becoming increasingly useful tools for the analysis of gene expression and single nucleotide polymorphisms (SNPs). Here, we present a method that permits the manufacture of microarrays from non-modified oligonucleotides on a poly carbodiimide-coated glass surface by UV-irradiation. The use of UV-irradiation facilitates an increase in the level of signal intensity, but it does not affect signal discrimination by the oligonucleotides immobilized on the surface. The signal intensity obtained for an array fabricated using non-modified oligonucleotides with UV-irradiation is ~7-fold greater than that without UV-irradiation. The detection of SNPs was tested to ascertain whether this technique could discriminate specific hybridization signals without causing significant UV-irradiation-induced damage to the immobilized oligonucleotides. We found that this immobilization method provides greater hybridization signals and a better match/mismatch ratio of SNPs than do the established aminosilane techniques. Application of this technology to manufacturing DNA microarrays for sequence analysis is discussed.     

On the conjugate hand reaction: Communication IV. Changing rates of conjugate hand reaction to sound and light modality signals upon surgical interventions in the subcortical brain structures in humans

The results of the study on conjugate reaction time (RT) of hands (the time of simple mental reaction) of 16 patients with Parkinsonism, cerebral palsy, and spastic torticollis before and after surgery are presented. Conjugation of the left hand and right hand RTs to sound and light modality signals with a warning signal has been analyzed to detect the morphological structures that influence the conjugate reaction. In some of the patients, no disturbance of RT conjugation was shown; in other patients, the coefficient of correlation used for assessment of the left and right hand RT conjugation significantly changed. The coefficient of correlation between the left hand and right hand RTs decreased in response either to the sound signal or simultaneously to the sound and light signals. Disturbances of the conjugate hand reaction were observed in the case of ventral-lateral thalamotomy, subthalamotomy, and pallidotomy.     

Integration of light and plastid signals

Light and plastid signals promote chloroplast biogenesis and are among the most potent inducers and repressors of photosynthesis-related gene expression, respectively. These signals can be likened to a 'gas and brake system' that promotes efficient chloroplast biogenesis and function. Recent findings indicate that a particular plastid signal can 'rewire' a light signaling network, converting it from an inducer into a repressor of particular photosynthesis-related genes. Therefore, a plastid signal appears to be an endogenous regulator of light signaling rather than a signal acting independently from light. This integration of light and plastid signals may allow plants to proactively manage chloroplast dysfunction when performing chloroplast biogenesis and maintenance in adverse light conditions.     

Wavelet Representation of the Corneal Pulse for Detecting Ocular Dicrotism

Purpose

To develop a reliable and powerful method for detecting the ocular dicrotism from non-invasively acquired signals of corneal pulse without the knowledge of the underlying cardiopulmonary information present in signals of ocular blood pulse and the electrical heart activity.

Methods

Retrospective data from a study on glaucomatous and age-related changes in corneal pulsation [PLOS ONE 9(7),(2014):e102814] involving 261 subjects was used. Continuous wavelet representation of the signal derivative of the corneal pulse was considered with a complex Gaussian derivative function chosen as mother wavelet. Gray-level Co-occurrence Matrix has been applied to the image (heat-maps) of CWT to yield a set of parameters that can be used to devise the ocular dicrotic pulse detection schemes based on the Conditional Inference Tree and the Random Forest models. The detection scheme was first tested on synthetic signals resembling those of a dicrotic and a non-dicrotic ocular pulse before being used on all 261 real recordings.

Results

A detection scheme based on a single feature of the Continuous Wavelet Transform of the corneal pulse signal resulted in a low detection rate. Conglomeration of a set of features based on measures of texture (homogeneity, correlation, energy, and contrast) resulted in a high detection rate reaching 93%.

Conclusion

It is possible to reliably detect a dicrotic ocular pulse from the signals of corneal pulsation without the need of acquiring additional signals related to heart activity, which was the previous state-of-the-art. The proposed scheme can be applied to other non-stationary biomedical signals related to ocular dynamics.     

Rapid photoperiodic responses in Japanese quail: is daylength measurement based upon a circadian system?

Experimental photoperiods, presented either once only or repeatedly, were used to assess the oscillatory and hourglass properties of the photoperiodic clock in Japanese quail. Gonadectomized quail on 8-hr daylengths respond to a single skeleton photoperiod consisting of two 8-hr light pulses separated by 2 hr of darkness (i.e., LDLD 8:2:8:6) with a marked increase in secretion rate of luteinizing hormone (LH). This response suggests that the second light pulse interacts with a "photoinducible phase" (phi i) lying some 10-16 hr from "dawn" (start of the first light pulse). If, however, groups of quail maintained on 8-hr daylengths are transferred to continuous darkness (DD), and the position of the phi i is sought by a single 8-hr light pulse applied at various times on the first or third day of DD, then an increase in circulating LH is, at best, barely detectable. It would appear that a strongly responsive phi i does not recur rhythmically in DD. Instead, the light pulse apparently acts primarily as a "dawn" signal that triggers a single cycle of photoinducibility, since a second 8-hr light pulse, placed to begin 2 hr after the end of the first, induces a large increase in plasma LH. Similar results are obtained if any single 8-hr light pulse presented to animals held in darkness is preceded, 10 hr earlier, by a short "dawn" light signal. Such dawn signals can be effective when very short; a pulse of only 30 sec can cause a subsequent phi i. The dawn pulse is effective at any circadian phase and leads to a single cycle in photoinducibility. In contrast, a much longer light pulse (perhaps not less than 4 hr) is needed to interact with phi i if significant gonadotropin secretion is to be stimulated. In confirmation of the findings described above, we found that Nanda-Hammer lighting schedules have remarkably little effect in stimulating gonadotropin secretion in gonadectomized quail. There is, for example, a very marked difference between the effectiveness of "resonating" schedules such as LD 6:6, which stimulates a high LH secretion rate since each "inductive" light pulse is preceded by an appropriate "dawn" signal, and a theoretically effective schedule such as LD 6:30, which induces a very small response by comparison. Such schedules (even theoretically noninductive ones) can, however, be made very highly inductive if alternate light pulses are preceded by an appropriately positioned 15-min light pulse to act as "dawn."     

Dynalets: A new method for modelling and compressing biological signals. Applications to physiological and molecular signals

The biological information coming from electrophysiologic sensors like ECG, pulse sensor or from molecular signal devices like NMR spectrometry has to be visualized and manipulated in a compressed way for an efficient medical use by clinicians, if stored in scientific data bases or in personalized patient records repositories. Here, we define a new transform called Dynalet based on Liénard ordinary differential equations susceptible to model the mechanism at the source of the studied signal, and we propose to apply this new technique first to the modelling and compression of real biological periodic signals like ECG and pulse rhythm. We consider that the cardiovascular activity results from the summation of cellular oscillators located in the cardiac sinus node and we show that, as a result, the van der Pol oscillator (a particular Liénard system) fits well the ECG signal and the pulse signal. The reconstruction of the original signal (pulse or ECG) using Dynalet transform is then compared with that of Fourier, counting the number of parameters to be set for obtaining an expected signal-to-noise ratio. Then, we apply the Dynalet transform to the modelling and compression of molecular spectra obtained by protein NMR spectroscopy. The reconstruction of the original signal (peak) using Dynalet transform is again compared with that of Fourier. After reconstructing visually the peak, we propose to periodize the signal and give it to hear, the whole process being called the protein “stethoscope”.