High Beat Frequency Research Articles

Kinematic characteristics of calanoid copepod appendage motion

Abstract Propulsion by copepods requires high-speed video and intensive analyses. Routine smooth swimming is compared with small relocation jumps and escape reactions, relying on kinematics analyses of appendage movement. In this study, we used high-speed video at 1200 frames per second to determine the kinematic parameters of three types of swimming in Eurytemora affinis females: routine steady swimming, small relocation jumps and escape reactions. The average speed varied in the range 0.30–0.82 cm s−1 during steady routine swimming. This value increased to 4.48 ± 1.01 cm s−1 during small relocation jumps, and reached 21.94 ± 2.68 cm s−1 during escape reactions. The small angular amplitude (40–50°) and the very high beat frequency (63–80 Hz) of the cephalic appendages during routine swimming indicate that E. affinis is a feeding-current feeder. The comparison of the angular displacements of the antennules, thoracic legs and abdomen with respect to the movement speed demonstrates that, both in the case of small relocation jumps and during escape reactions, the main propulsive role is played by the thoracic swimming legs and abdomen.

Structural Analysis of Sperm Centrioles Using N-STORM.

A prominent technical barrier when imaging swimming sperm is capturing a singular sperm cell's head and tail position simultaneously at a high resolution to understand their relationship in different stages of the sperm tail beating cycle. This is due to the sperm's high beating frequency, rotational movement, and the large difference in diameter between the head and tail. These intricacies increase the complexity of determining the position of a dynamic subcellular structure in the sperm neck, such as the centriole. We have developed a way to obtain this information by snap freezing mobile sperm at different stages of the sperm tail beating cycle and then analyzing them with super-resolution microscopy. This method captures the position of both the sperm head and tail at the microscale and centriolar substructure details at the nanoscale. This chapter describes the detailed procedures for the selection, preparation, antibody staining, 3D N-STORM imaging, and image quantification of bovine spermatozoa.

3D+t feature-based descriptor for unsupervised flagellar human sperm beat classification.

Human spermatozoa must swim through the female reproductive tract, where they undergo a series of biochemical and biophysical reactions called capacitation, a necessary step to fertilize the egg. Capacitation promotes changes in the motility pattern. Historically, a two-dimensional analysis has been used to classify sperm motility and clinical fertilization studies. Nevertheless, in a natural environment sperm motility is three-dimensional (3D). Imaging flagella of freely swimming sperm is a difficult task due to their high beating frequency of up to 25 Hz. Very recent studies have described several sperm flagellum 3D beating features (curvature, torsion, asymmetries, etc.). However, up to date, the 3D motility pattern of hyperactivated spermatozoa has not been characterized. The main difficulty in classifying these patterns in 3D is the lack of a ground truth reference since differences in flagellar beat patterns are very difficult to assess visually. Moreover, only around 10-20% of induced to capacitate spermatozoa are truly capacitated, i.e., hyperactivated. We used an image acquisition system that can acquire, segment, and track spermatozoa flagella in 3D+t. In this work, we propose an original three-dimensional feature vector formed by ellipses describing the envelope of the 3D+t spatio-temporal flagellar sperm motility patterns. These features allowed compressing an unlabeled 3D+t dataset to separate hyperactivated cells from others (capacitated from non-capacitated cells) using unsupervised hierarchical clustering. Preliminary results show three main clusters of flagellar motility patterns. The first principal component of these 3D flagella measurements correlated with 2D OpenCASA head determinations as a first approach to validate the unsupervised classification, showing a reasonable correlation coefficient near to 0.7. Clinical relevance- The novelty of this work is defining a 3D+t feature-based descriptor consisting of a set of ellipses enveloping the flagellar motion of human sperm for its unsu-pervised classification. This is a new promising tool to determine the viability of human sperm to fertilize the egg.

Engineering rotating apical-out airway organoid for assessing respiratory cilia motility

SummaryMotile cilia project from the airway apical surface and directly interface with inhaled external environment. Owing to cilia’s nanoscale dimension and high beating frequency, quantitative assessment of their motility remains a sophisticated task. Here we described a robust approach for reproducible engineering of apical-out airway organoid (AOAO) from a defined number of cells. Propelled by exterior-facing cilia beating, the mature AOAO exhibited stable rotational motion when surrounded by Matrigel. We developed a computational framework leveraging computer vision algorithms to quantify AOAO rotation and correlated it with the direct measurement of cilia motility. We further established the feasibility of using AOAO rotation to recapitulate and measure defective cilia motility caused by chemotherapy-induced toxicity and by CCDC39 mutations in cells from patients with primary ciliary dyskinesia. We expect our rotating AOAO model and the associated computational pipeline to offer a generalizable framework to expedite the modeling of and therapeutic development for genetic and environmental ciliopathies.

Swimming Kinematics of Cyprids of the Barnacle Balanus glandula.

Larvae of barnacles typically pass through naupliar and cyprid planktonic stages before settlement and metamorphosis. As the final larval stage, cyprids swim much faster than nauplii and in turbulent fluid environments with high shears as they seek habitat. Cyprids swim with six pairs of reciprocating thoracic appendages and use two anterior antennules during settlement. Our understanding of how thoracic appendages generate movement is limited due to short stroke intervals (∼5 ms) that impede observations of the shape and trajectory of appendages. Here, we used high-speed videography to observe both free-swimming and tethered cyprids of the intertidal acorn barnacle Balanus glandula to produce a comprehensive description of thoracic appendage swimming kinematics. Cyprids used a drag-based method of swimming: their six pairs of thoracic appendages moved through metachronal power strokes and synchronous recovery strokes similar to the thoracopod motions in calanoid copepods during escape swimming. During the power stroke, plumose setae on each appendage pair spread laterally into a high surface area and high drag paddle composed of a meshwork of fused setules. This interconnected setal array collapsed into a low surface area and low drag shape during the recovery stroke. These effective swimming appendages allowed cyprids to move upward at an average speed of 1.4 cm/s (∼25 body lengths/s) with an average beat frequency of 16 beats/s, and reach an instantaneous velocity of up to 6 cm/s. Beat frequency of the thoracic appendages was significantly associated with speed, with higher beat frequencies indicating faster swimming speed. At their average speed, cyprids moved at the intermediate Reynolds number of ∼10, in which both viscous and inertial forces affected movement. Cyprids could alter swimming direction by sweeping the posterior-most appendage pair to one side and beating the remaining thoracic appendages synchronously through the power stroke with greater motion on the outside of their turn. These results greatly enhance our understanding both of cyprid motility and how small planktonic organisms can use swimming appendages with fused setule arrays to reach high swimming speeds and affect directional changes.

FMCW LiDAR System to Reduce Hardware Complexity and Post-Processing Techniques to Improve Distance Resolution.

As the autonomous driving technology develops, research on related sensors is also being actively conducted. One system that is widely used today uses a light source with a wavelength in the 905 nm band for the pulse Light Detection And Ranging (LiDAR) system. This has the disadvantages of being harmful to the human eye and in making digital signal processing difficult at high sampling rates. The Frequency Modulated Continuous Wave (FMCW) LiDAR system has been proposed as an alternative. However, the FMCW LiDAR is formed with a high beat frequency by a method different from that of the FMCW Radar, which causes a hardware burden on the FFT (Fast Fourier Transform) module for interpreting the beat frequency information. In this paper, the FFT module that may occur in the FMCW LiDAR using Digital Down Convert (DDC) technology is extracted at 256 points, which is times smaller than the existing 8192 points, and the beat frequency is 0 to 50 m at 3 cm intervals. As a result of generating and restoring the distance, the performance of 0.03 m Root Mean Square Error (RMSE) compared to the conventional one was confirmed. In this process, the hardware module was directly mounted and verified on the FPGA. In the case of the Simple Threshold-Constant False Alarm Rate (ST-CFAR) provided, the RMSE was measured by generating beat frequencies from 0 to 50 m at 1 cm intervals, and as a result, the result of 0.019 m was confirmed at 0.03 m in the past.

Digital frequency offset-locked He–Ne laser system with high beat frequency stability, narrow optical linewidth and optical fibre output

We report on the enhancement of a previously-presented heterodyne laser source on the basis of two phase-locked loop (PLL) frequency coupled internal-mirror He–Ne lasers. Our new system consists of two digitally controlled He–Ne lasers with slightly different wavelengths, and offers high-frequency stability and very narrow optical linewidth. The digitally controlled system has been realized by using a FPGA controller and transconductance amplifiers. The light of both lasers was coupled into separate fibres for heterodyne interferometer applications. To enhance the laser performance we observed the sensitivity of both laser tubes to electromagnetic noise from various laser power supplies and frequency control systems. Furthermore, we describe how the linewidth of a frequency-controlled He–Ne laser can be reduced during precise frequency stabilisation. The digitally controlled laser source reaches a standard beat frequency deviation of less than 20 Hz (with 1 s gate time) and a spectral full width at half maximum (FWHM) of the beat signal less than 3 kHz. The laser source has enough optical output power to serve a fibre-coupled multi axis heterodyne interferometer. The system can be adjusted to output beat frequencies in the range of 0.1 MHz–20 MHz.

Biomechanics of swimming in developing larval fish.

Most larvae of bony fish are able to swim almost immediately after hatching. Their locomotory system supports several vital functions: fish larvae make fast manoeuvres to escape from predators, aim accurately during suction feeding and may migrate towards suitable future habitats. Owing to their small size and low swimming speed, larval fish operate in the intermediate hydrodynamic regime, which connects the viscous and inertial flow regimes. They experience relatively strong viscous effects at low swimming speeds, and relatively strong inertial effects at their highest speeds. As the larvae grow and increase swimming speed, a shift occurs towards the inertial flow regime. To compensate for size-related limitations on swimming speed, fish larvae exploit high tail beat frequencies at their highest speeds, made possible by their low body inertia and fast neuromuscular system. The shifts in flow regime and body inertia lead to changing functional demands on the locomotory system during larval growth. To reach the reproductive adult stage, the developing larvae need to adjust to and perform the functions necessary for survival. Just after hatching, many fish larvae rely on yolk and need to develop their feeding systems before the yolk is exhausted. Furthermore, the larvae need to develop and continuously adjust their sensory, neural and muscular systems to catch prey and avoid predation. This Review discusses the hydrodynamics of swimming in the intermediate flow regime, the changing functional demands on the locomotory system of the growing and developing larval fish, and the solutions that have evolved to accommodate these demands.

The effect of beat frequency on eye movements during free viewing

External periodic stimuli entrain brain oscillations and affect perception and attention. It has been shown that background music can change oculomotor behavior and facilitate detection of visual objects occurring on the musical beat. However, whether musical beats in different tempi modulate information sampling differently during natural viewing remains to be explored. Here we addressed this question by investigating how listening to naturalistic drum grooves in two different tempi affects eye movements of participants viewing natural scenes on a computer screen. We found that the beat frequency of the drum grooves modulated the rate of eye movements: fixation durations were increased at the lower beat frequency (1.7Hz) as compared to the higher beat frequency (2.4Hz) and no music conditions. Correspondingly, estimated visual sampling frequency decreased as fixation durations increased with lower beat frequency. These results imply that slow musical beats can retard sampling of visual information during natural viewing by increasing fixation durations.

DFB fiber laser static strain sensor based on beat frequency interrogation with a reference fiber laser locked to a FBG resonator.

We report on a high-resolution static strain sensor developed with distributed feedback (DFB) fiber laser. A reference FBG resonator is used for temperature compensation. Locking another independent fiber laser to the resonator using the Pound-Drever-Hall technique results in a strain power spectral density better than Sε(f) = (4.6 × 10-21) ε2/Hz in the frequency range from 1 Hz to 1 kHz, corresponding to a minimum dynamic strain resolution of 67.8 pε/√Hz. This frequency stabilized fiber laser is proposed to interrogate the sensing DFB fiber laser by the beat frequency principle. As a reasonable DFB fiber laser setup is realized, a narrow beat frequency line-width of 3.23 kHz and a high beat frequency stability of 0.036 MHz in 15 minutes are obtained in the laboratory test, corresponding to a minimum static strain resolution of 270 pε. This is the first time that a sub-0.5 nε level for static strain measurement using DFB fiber laser is demonstrated.

Imaging of the 3D dynamics of flagellar beating in human sperm.

The study of the mechanical and environmental factors that regulate a fundamental event such as fertilization have been subject of multiple studies. Nevertheless, the microscopical size of the spermatozoa and the high beating frequency of their flagella (up to 20 Hz) impose a series of technological challenges for the study of the mechanical factors implicated. Traditionally, due to the inherent characteristics of the rapid sperm movement, and to the technological limitations of microscopes (optical or confocal) to follow in three dimensions (3D) their movement, the analysis of their dynamics has been studied in two dimensions, when the head is confined to a surface. Flagella propel sperm and while their head can be confined to a surface, flagellar movement is not restricted to 2D, always displaying 3D components. In this work, we present a highly novel and useful tool to analyze sperm flagella dynamics in 3D. The basis of the method is a 100 Hz oscillating objective mounted on a bright field optical microscope covering a 16 microns depth space at a rate of ~ 5000 images per second. The best flagellum focused subregions were associated to their respective Z real 3D position. Unprecedented graphical results making evident the 3D movement of the flagella are shown in this work and supplemental material illustrating a 3D animation using the obtained experimental results is also included.

高功率可调谐双频激光全光纤放大实验研究

高功率可调谐双频激光全光纤放大实验研究

Temperature and fatigue effect on the maximum swimming speed of jack mackerel Trachurus japonicus

Swimming performance of jack mackerel Trachurus japonicus (18.2 ± 0.8 cm fork length (FL), n = 185) was examined in a flume tank by measuring the stride length at low and high tail beat frequencies with electromyogram monitoring and a muscle twitch experiment. Stride length was analyzed by monitoring the tail beat frequency according to the swimming speed at different temperatures of 10, 15 and 22 °C. In the electromyographic observations, the initiation of ordinary muscle activity occurred between 71.4 and 99.6 cm/s, that is 3.7 to 5.3 FL/s, when the tail beat frequency was over 6 Hz. The swimming speeds increased rectilinearly with the tail beat frequency at each water temperature both for the low and high tail beat frequency. Lower stride length was observed at the lowest temperature (10 °C) tested. The forced swimming exercise significantly affected the muscle contraction time to become longer than the control fish, which indicated a reduction of the maximum swimming speed performance.

Phylogenetic and Morphological Diversity of Novel Soil Cercomonad Species with a Description of Two New Genera (Nucleocercomonas and Metabolomonas)

Cercomonads are important components of microbial food webs in soils and aquatic sediments. Here, we investigated the general morphology, behaviour, life cycle and 18S rDNA phylogeny of cercomonad cultures from a German grassland soil habitat. We describe ten new species including two new genera from 23 strains. Three Cercomonas, two Eocercomonas and three Paracercomonas species are described. Based on large phylogenetic distance and distinct morphology, we erect two novel clade B genera near the root of the cercomonad tree. Nucleocercomonas nov. gen. bears a number of characters unusual for cercomonads: Its anterior flagellum is extremely long, it mostly does not glide, and in its most frequent life stage the cell body does not attach to the substratum, but produces unattached pseudopodia. Furthermore, it has a unique nucleus with a peripheral nucleolus that attaches to the nuclear envelope opposite the basal body connection. Metabolomonas nov. gen. is extremely metabolic. It is characterized by a very high beating frequency of the anterior flagellum, fast gliding, rapid changes in shape and strong cytoplasmic streams. A new genus Brevimastigomonas is erected for the previously described species Paracercomonas anaerobica. The general morphology of cercomonad species often does not correspond with their phylogenetic position: closely related species may have a very different morphology.

Frenetic activation of fish spermatozoa flagella entails short‐term motility, portending their precocious decadence

In most species, fish spermatozoa activate their motility on contact with the external medium (sea or fresh water depending of their reproductive habitat). Their flagella immediately develop waves propagated at high beat frequency (up to 70 beats s(-1)), which propel these sperm cells at high velocity (6-10 mm min(-1)), but for a quite short period of time, usually limited to minutes. Their specific inability to restore their energy content (mostly adenosine triphosphate) fast enough relatively to their high rate of energy consumption by flagellar contributes mainly to the activity arrest of motility, as the spermatozoa need to rely on early accumulated energy prior to activation. This review of the published data explains the present understanding of physico-chemical mechanisms by which flagellar motility is activated (mostly through osmotic and ionic regulation) and then propels sperm cells at speed. It aims also to describe the gradual arrest of their motility much of which occurs within a few minutes.

Frequency jitter and spectral width of an injection-seeded Q-switched Nd:YAG laser for a Doppler wind lidar

The design of a 50 Hz single longitudinal mode, diode-pumped and frequency-tripled Nd:YAG master oscillator power amplifier is described, and the first measurements of output parameters are presented. The laser oscillator is injection-seeded by a tuneable monolithic Nd:YAG ring laser and frequency stabilized by minimising the Q-switch build-up time. The laser system will be an integral part of an airborne instrument demonstrator for a first satellite based Doppler wind lidar to measure vertical profiles of one component of the atmospheric wind vector. This paper focuses on the investigation of the frequency jitter and the linewidth of the laser, which are measured on a pulse-to-pulse basis. For this purpose a compact, high accuracy beat frequency monitoring system has been developed at DLR. By operating the amplifier stage at half the repetition rate (50 Hz) of the oscillator, we could reduce the frequency stability from 10 MHz (rms) to 1.3 MHz (rms) (over a 14 s period). We have determined a mean linewidth of 15 MHz (FWHM) at 1064 nm. These measured laser parameters enable wind velocity measurements in the atmosphere (0–15 km) at an accuracy of 1 to 2 m/s.

Hyperactivation is the mode conversion from constant‐curvature beating to constant‐frequency beating under a constant rate of microtubule sliding

Flagellar beating of hyperactivated golden hamster spermatozoa was analyzed in detail using digital image analysis and was compared to that of nonhyperactivated (activated) spermatozoa in order to understand the change in flagellar beating during hyperactivation and the active microtubule sliding that brought about the change in flagellar beating. Hyperactivated flagellar beating, which was characterized by a sharp bend in the proximal midpiece and low beat frequency, was able to alter the waveform with little change in beat frequency (constant-frequency beating), whereas activated flagellar beating, which was characterized by a slight bend in the proximal midpiece and high beat frequency, was able to alter beat frequency with little change in the waveform (constant-curvature beating). These results demonstrate that flagellar beating of hyperactivated and activated spermatozoa were essentially different modes and that hyperactivation was the mode conversion from constant-curvature beating to constant-frequency beating. Detailed analysis of flagellar bends revealed that the increase in curvature in the proximal midpiece during hyperactivation was due to the increase in total length of microtubule sliding in a nearly straight region between bends, while the rate of microtubule sliding remained almost constant.

Modeling of the Kinetics of Intramolecular Processes and Transient Spectra with Allowance for Nonradiative Transitions

It has been shown that the kinetics of intramolecular processes and time-resolved spectra with allowance for the quantum beats of the resonant states of isomers or isolated subsystems of levels of one isomeric form can be described with the use of a molecular model interpreting the effect of beats as a nonradiative transition. We have obtained an expression for the nonradiative transition probability, which is directly proportional to the beat frequency and depends oscillatorily on time, thus modeling the effect of beats. The parameter of the molecular system model is the beat frequency directly related to the parameter characterizing the intramolecular interisomeric interactions (the corresponding nondiagonal element of the energy matrix) rather than the value of the nonradiative transition probability. The character of the change in the level populations and, accordingly, in the band intensities in the spectra in the proposed model is in good agreement with the experiment, including the fine structure of the time dependences — oscillations of the line intensities. In analyzing the temporal experiment with a high resolution, it is necessary to take into account the instrument function leading to quantitative and qualitative changes in the time dependences. The traditional model of nonradiative transitions with a constant probability value has a very limited range of applicability — very high beat frequencies compared to the probability of optical transitions.

Glycolysis plays a major role for adenosine triphosphate supplementation in mouse sperm flagellar movement.

The mammalian sperm must be highly motile for a long time to fertilize a egg. It has been supposed that ATP required for sperm flagellar movement depends predominantly on mitochondrial respiration. We assessed the contribution of mitochondrial respiration to mouse sperm motility. Mouse sperm maintained vigorous motility with high beat frequency in an appropriate solution including a substrate such as glucose. The active sperm contained a large amount of ATP. When carbonyl cyanide m-chlorophenylhydrazone (CCCP) was applied to suppress the oxidative phosphorylation in mitochondria, the vigorous motility was maintained and the amount of ATP was kept at the equivalent level to that without CCCP. When pyruvate or lactate was provided instead of glucose, both sperm motility and the amount of ATP were high. However, they were drastically decreased when oxidative phosphorylation was suppressed by addition of CCCP. We also found that sperm motility could not be maintained in the presence of respiratory substrates when glycolysis was suppressed. 2-Deoxy-d-glucose (DOG) had no effect on mitochondrial respiration assessed by a fluorescent probe, 5,5',6,6'-tetrachloro-1,1',3,3'-tetraethylbenzimidazolylcarbocyanine iodide (JC-1), but, it inhibited motility and decreased ATP content when pyruvate or lactate were provided as substrates. The present results suggest that glycolysis has an unexpectedly important role in providing the ATP required for sperm motility throughout the length of the sperm flagellum.

Two-longitudinal-mode He-Ne laser for heterodyne interferometers to measure displacement.

We propose a new configuration for a high-resolution heterodyne interferometer that employs a two-longitudinal-mode He-Ne laser with an intermode beat frequency of 600-1000 MHz. The high beat frequency is downconverted to 5 MHz such that the phase change of the interferometer output is precisely measured with a displacement resolution of 0.1 nm. A thermal control scheme is adopted to stabilize the cavity length of the He-Ne plasma tube such that a frequency stability of 2 parts in 10(9) is obtained by suppression of frequency drifts caused by t he phenomena of frequency pulling and polarization anisotropy. This two-longitudinal-mode He-Ne laser yields a high output power of 2.0 mW, which permits multiple measurements of as many as 10 machine axes simultaneously.