cAMP and the Fibrous Sheath Protein CABYR (Ca2+-Binding Tyrosine-Phosphorylation-Regulated Protein) Is Required for 4D Sperm Movement.

A new life starts with successful fertilization whereby one sperm from a pool of millions fertilizes the oocyte. Sperm motility is one key factor for this selection process, which depends on a coordinated flagellar movement. The flagellar beat cycle is regulated by Ca2+ entry via CatSper, cAMP, Mg2+, ADP and ATP. This study characterizes the effects of these parameters for 4D sperm motility, especially for flagellar movement and the conserved clockwise (CW) path chirality of murine sperm. Therefore, we use detergent-extracted mouse sperm and digital holographic microscopy (DHM) to show that a balanced ratio of ATP to Mg2+ in addition with 18 µM cAMP and 1 mM ADP is necessary for controlled flagellar movement, induction of rolling along the long axis and CW path chirality. Rolling along the sperm's long axis, a proposed mechanism for sperm selection, is absent in sea urchin sperm, lacking flagellar fibrous sheath (FS) and outer-dense fibers (ODFs). In sperm lacking CABYR, a Ca2+-binding tyrosine-phosphorylation regulated protein located in the FS, the swim path chirality is preserved. We conclude that specific concentrations of ATP, ADP, cAMP and Mg2+ as well as a functional CABYR play an important role for sperm motility especially for path chirality.

Multi-ciliated microswimmers-metachronal coordination and helical swimming.

The dynamics and motion of multi-ciliated microswimmers with a spherical body and a small number N (with [Formula: see text]) of cilia with length comparable to the body radius, is investigated by mesoscale hydrodynamics simulations. A metachronal wave is imposed for the cilia beat, for which the wave vector has both a longitudinal and a latitudinal component. The dynamics and motion is characterized by the swimming velocity, its variation over the beat cycle, the spinning velocity around the main body axis, as well as the parameters of the helical trajectory. Our simulation results show that the microswimmer motion strongly depends on the latitudinal wave number and the longitudinal phase lag. The microswimmers are found to swim smoothly and usually spin around their own axis. Chirality of the metachronal beat pattern generically generates helical trajectories. In most cases, the helices are thin and stretched, i.e., the helix radius is about an order of magnitude smaller than the pitch. The rotational diffusion of the microswimmer is significantly smaller than the passive rotational diffusion of the body alone, which indicates that the extended cilia contribute strongly to the hydrodynamic radius. The swimming velocity is found to increase with the cilia number N with a slightly sublinear power law, consistent with the behavior expected from the dependence of the transport velocity of planar cilia arrays on the cilia separation.

Chiral-filament self-assembly on curved manifolds.

Rod-like and banana-shaped proteins, like BAR-domain proteins and MreB proteins, adsorb on membranes and regulate the membrane curvature. The formation of large filamentous complexes of these proteins plays an important role in cellular processes like membrane trafficking, cytokinesis and cell motion. We propose a simplified model to investigate such curvature-dependent self-assembly processes. Anisotropic building blocks, modeled as trimer molecules, which have a preferred binding site, interact via pair-wise Lennard-Jones potentials. When several trimers assemble, they form an elastic ribbon with an intrinsic curvature and twist, controlled by bending and torsional rigidity. For trimer self-assembly on the curved surface of a cylindrical membrane, this leads to a preferred spatial orientation of the ribbon. We show that these interactions can lead to the formation of helices with several windings around the cylinder. The emerging helix angle and pitch depend on the rigidities and the intrinsic curvature and twist values. In particular, a well-defined and controllable helix angle emerges in the case of equal bending and torsional rigidity. The dynamics of filament growth is characterized by three regimes, in which filament length increases with the power laws tz in time, with z≃ 3/4, z = 1/2, and z = 0 for short, intermediate, and long times, respectively. A comparison with the solutions of the Smoluchowski aggregation equation allows the identification of the underlying mechanism in the short-time regime as a crossover from size-independent to diffusion-limited aggregation. Thus, helical structures, as often observed in biology, can arise by self-assembly of anisotropic and chiral proteins.

Interferometric delay tracking for low-noise Mach-Zehnder-type scanning measurements.

Precise delay control is of paramount importance in optical pump-probe measurements. Here, we report on a high-precision delay tracking technique for mechanical scanning measurements in a Mach-Zehnder interferometer configuration. The setup employs a 1.55-µm continuous-wave laser beam propagating along the interferometer arms. Sinusoidal phase modulation at 30 MHz, and demodulation of the interference signal at the fundamental frequency and its second harmonic, enables delay tracking with sampling rates of up to 10 MHz. At an interferometer arm length of 1 m, root-mean-square error values of the relative delay tracking below 10 attoseconds for both stationary and mechanically scanned (0.2 mm/s) operation are demonstrated. By averaging several scans, a precision of the delay determination better than 1 as is reached. We demonstrate this performance with a mechanical chopper periodically interrupting one of the interferometer arms, which opens the door to the combination of high-sensitivity lock-in detection with (sub-)attosecond-precision relative delay determination.

Frequency-modulated atomic force microscopy operation by imaging at the frequency shift minimum: the dip-df mode.

In frequency modulated non-contact atomic force microscopy, the change of the cantilever frequency (Δf) is used as the input signal for the topography feedback loop. Around the Δf(z) minimum, however, stable feedback operation is challenging using a standard proportional-integral-derivative (PID) feedback design due to the change of sign in the slope. When operated under liquid conditions, it is furthermore difficult to address the attractive interaction regime due to its often moderate peakedness. Additionally, the Δf signal level changes severely with time in this environment due to drift of the cantilever frequency f0 and, thus, requires constant adjustment. Here, we present an approach overcoming these obstacles by using the derivative of Δf with respect to z as the input signal for the topography feedback loop. Rather than regulating the absolute value to a preset setpoint, the slope of the Δf with respect to z is regulated to zero. This new measurement mode not only makes the minimum of the Δf(z) curve directly accessible, but it also benefits from greatly increased operation stability due to its immunity against f0 drift. We present isosurfaces of the Δf minimum acquired on the calcite CaCO3(101̅4) surface in liquid environment, demonstrating the capability of our method to image in the attractive tip-sample interaction regime.

Photothermal excitation setup for a modified commercial atomic force microscope.

High-resolution imaging in liquids using frequency modulation atomic force microscopy is known to suffer from additional peaks in the resonance spectrum that are unrelated to the cantilever resonance. These unwanted peaks are caused by acoustic modes of the liquid and the setup arising from the indirect oscillation excitation by a piezoelectric transducer. Photothermal excitation has been identified as a suitable method for exciting the cantilever in a direct manner. Here, we present a simple design for implementing photothermal excitation in a modified Multimode scan head from Bruker. Our approach is based on adding a few components only to keep the modifications as simple as possible and to maintain the low noise level of the original setup with a typical deflection noise density of about 15 fm/√[Hz] measured in aqueous solution. The success of the modification is illustrated by a comparison of the resonance spectra obtained with piezoelectric and photothermal excitation. The performance of the systems is demonstrated by presenting high-resolution images on bare calcite in liquid as well as organic adsorbates (Alizarin Red S) on calcite with simultaneous atomic resolution of the underlying calcite substrate.

Convolution theory for dynamic systems: a bottom-up approach to the viscoelasticity of polymeric networks.

Biological materials such as the cytoskeleton are characterized by remarkable viscoelastic properties and therefore represent the subject of numerous micro- and macrorheological experimental studies. By generalizing the previously introduced dynamic convolution theory (DCT) to two dimensions, we devise a bottom-up approach for the viscoelastic properties of extended, crosslinked semiflexible polymer networks. Brownian dynamics (BD) simulations serve to determine the dynamic linear self- and cross-response properties of isolated semiflexible polymers to externally applied forces and torques; these response functions are used as input to the DCT. For a given network topology, the frequency-dependent response of the network subject to a given external force/torque distribution is calculated via the DCT allowing to resolve both micro- and macrorheological properties of the networks. A mapping on continuum viscoelastic theory yields the corresponding viscoelastic bulk moduli. Special attention is drawn to the flexibility of crosslinkers, which couple angular degrees of freedom at the network nodes and which are found to sensitively affect the resulting rheological properties of the polymeric meshwork.

Towards dynamical network biomarkers in neuromodulation of episodic migraine.

Computational methods have complemented experimental and clinical neurosciences and led to improvements in our understanding of the nervous systems in health and disease. In parallel, neuromodulation in form of electric and magnetic stimulation is gaining increasing acceptance in chronic and intractable diseases. In this paper, we firstly explore the relevant state of the art in fusion of both developments towards translational computational neuroscience. Then, we propose a strategy to employ the new theoretical concept of dynamical network biomarkers (DNB) in episodic manifestations of chronic disorders. In particular, as a first example, we introduce the use of computational models in migraine and illustrate on the basis of this example the potential of DNB as early-warning signals for neuromodulation in episodic migraine.

Thermal noise limit for ultra-high vacuum noncontact atomic force microscopy.

The noise of the frequency-shift signal Δf in noncontact atomic force microscopy (NC-AFM) consists of cantilever thermal noise, tip-surface-interaction noise and instrumental noise from the detection and signal processing systems. We investigate how the displacement-noise spectral density d(z) at the input of the frequency demodulator propagates to the frequency-shift-noise spectral density d(Δ) (f) at the demodulator output in dependence of cantilever properties and settings of the signal processing electronics in the limit of a negligible tip-surface interaction and a measurement under ultrahigh-vacuum conditions. For a quantification of the noise figures, we calibrate the cantilever displacement signal and determine the transfer function of the signal-processing electronics. From the transfer function and the measured d(z), we predict d(Δ) (f) for specific filter settings, a given level of detection-system noise spectral density d(z) (ds) and the cantilever-thermal-noise spectral density d(z) (th). We find an excellent agreement between the calculated and measured values for d(Δ) (f). Furthermore, we demonstrate that thermal noise in d(Δ) (f), defining the ultimate limit in NC-AFM signal detection, can be kept low by a proper choice of the cantilever whereby its Q-factor should be given most attention. A system with a low-noise signal detection and a suitable cantilever, operated with appropriate filter and feedback-loop settings allows room temperature NC-AFM measurements at a low thermal-noise limit with a significant bandwidth.

Surface modification of luminescent lanthanide phosphate nanorods with cationic "Quat-primer" polymers.

"Quat-primer" polymers bearing cationic groups were investigated as a surface modifier for Tb-doped cerium phosphate green-emitting fluorescent nanorods (NRs). The NRs were synthesized by a microwave process without using any complex agents or ligands and were characterized with different analytical tools such as X-ray diffraction, transmission electron microscopy, and fluorescence spectroscopy. Poly(ethyleneimine) partially quarternized with glycidyltrimethylammonium chloride was synthesized separately and characterized in detail. (1)H and (13)C NMR spectroscopic studies revealed that the quaternary ammonium group was covalently attached to the polymer. UV-vis spectroscopy was used to examine the stability of the colloidal dispersions of the bare NRs as well as the modified NRs. ζ potential, thermogravimetric analysis, and atomic force microscopy studies were carried out to confirm that the positively charged Quat-primer polymer is adsorbed on the negatively charged surface of the NRs, which results in high dispersion stability. Emission spectra of the modified NRs indicated that there was no interference of the Quat-primer polymer with the fluorescence behavior.

Clear signature of the (2 x 1) reconstruction of calcite (1014).

Calcite is a mineral of fundamental importance that plays a crucial role in many fields of research such as biomineralization, biomolecule adsorption, and reactivity as well as industrial and daily life applications. Consequently, the most stable cleavage plane of calcite has been studied extensively using both direct imaging techniques such as atomic force microscopy as well as spectroscopic and diffraction techniques. Several surface structures have been reported for the (1014) cleavage plane of calcite differing from the simple bulk-truncated structure and an ongoing controversy exists in literature whether the cleavage plane exhibits a (2 x 1) reconstruction or not. We study the (1014) cleavage plane using high-resolution noncontact atomic force microscopy (NC-AFM) under ultrahigh vacuum conditions and obtain a clear signature of the (2 x 1) reconstruction. This reconstruction is observed in very narrow tip-surface distance ranges only, explaining why in some experiments the reconstruction has been observed and in others not. Moreover, as all sample preparation is performed in ultrahigh vacuum, the possibility of the (2 x 1) reconstruction being adsorbate-induced appears rather unlikely. Additionally, tip-induced surface changes are ruled out as origin for the observed reconstruction either. In conclusion, our study suggests that the (2 x 1) reconstruction is a true surface property of the (1014) cleavage plane of calcite.

True atomic-resolution imaging of (1014) calcite in aqueous solution by frequency modulation atomic force microscopy.

Calcite (CaCO3) is one of the most abundant minerals on earth and plays an important role in a wide range of different fields including, for example, biomineralization and environmental geochemistry. Consequently, surface processes and reactions such as dissolution and growth as well as (macro)molecule adsorption are of greatest interest for both applied as well as fundamental research. An in-depth understanding of these processes requires knowledge about the detailed surface structure in its natural state which is quite often a liquid environment. We have studied the most stable cleavage plane of calcite under liquid conditions using frequency modulation atomic force microscopy. Using this technique, we achieved true atomic-resolution imaging, demonstrating the high-resolution capability of frequency modulation atomic force microscopy in liquids. We could reproduce contrast features reported before using contact mode atomic force microscopy, originating from the protruding oxygen atom of the carbonate groups. Besides this contrast, however, our results, indeed, indicate that we obtain more detailed structural information, revealing the calcium sublattice of the (1014) cleavage plane.