Data acquisition and imaging using wavelet transform: a new path for high speed transient force microscopy

This paper introduces a fundamentally new approach for dynamic AFM data acquisition and imaging based on applying the wavelet transform on the data stream from the photodetector.

Applying an updated method of markers to defining transient force impact under multiphase flowing

Numerical simulation of transient hydrodynamic forces from shaped gas cavities formed in liquid under active interaction of liquid and a jet source of high-temperature gas and intensive heat and mass transfer is performed. To solve the task, a method of coarse particle markers with, as opposed to the classical one, an additional stage, when moving boundaries of different media in cells with interfaces of these media are as if stitched, is updated. In addition, problems of inter-media heat and mass transfer by condensation and evaporation are simultaneously solved. The predicted results are compared with the experimental data. Validation and verification are performed by comparing the analysis results with the experimental data. The applicability of the updated method of coarse particle markers to defining transient force impact under multiphase flowing is demonstrated.

The transient force profile of low-speed droplet impact: measurements and model

The transient force exerted by a low-speed liquid droplet impinging onto a flat rigid surface is investigated experimentally. The measurements employ a high-sensitivity piezo-electric sensor, along with a high-speed camera, and cover four decades in droplet Reynolds number and greater than two decades in Weber number. Across these ranges, the peak of individual force profiles span from 3 mN to over 1300 mN. Once normalised, the force–time profiles support the existence of an inertially dominated self-similar regime. Within this regime, previous numerical and theoretical studies predict a $\sqrt{t}$ dependence of impact normal force during the initial pre-peak rise. While our measurements confirm this finding, they also indicate that, after the peak force the profiles exhibit an exponential decay. This long-time decay law suggests treatment of the momentum transport from the droplet using a lumped model. An observed linear dependence between the force and force decay rate supports this approach. The reason for the efficacy of treating this system via a lumped model apparently connects to the physics right at the surface that limit the rate of momentum transport from the droplet to the surface. This is explored by estimating the momentum transfer by solely using the deforming droplet shape, but under the condition of negligible momentum gradients within the droplet. The short- and long-time solutions are combined and the resulting model equation is shown to accurately cover the entire force–time profile.

Regulation of myofilament force and loaded shortening by skeletal myosin binding protein C

Myosin binding protein C (MyBP-C) is a 125–140-kD protein located in the C-zone of each half-thick filament. It is thought to be an important regulator of contraction, but its precise role is unclear. Here we investigate mechanisms by which skeletal MyBP-C regulates myofilament function using rat permeabilized skeletal muscle fibers. We mount either slow-twitch or fast-twitch skeletal muscle fibers between a force transducer and motor, use Ca2+ to activate a range of forces, and measure contractile properties including transient force overshoot, rate of force development, and loaded sarcomere shortening. The transient force overshoot is greater in slow-twitch than fast-twitch fibers at all Ca2+ activation levels. In slow-twitch fibers, protein kinase A (PKA) treatment (a) augments phosphorylation of slow skeletal MyBP-C (sMyBP-C), (b) doubles the magnitude of the relative transient force overshoot at low Ca2+ activation levels, and (c) increases force development rates at all Ca2+ activation levels. We also investigate the role that phosphorylated and dephosphorylated sMyBP-C plays in loaded sarcomere shortening. We test the hypothesis that MyBP-C acts as a brake to filament sliding within the myofilament lattice by measuring sarcomere shortening as thin filaments traverse into the C-zone during lightly loaded slow-twitch fiber contractions. Before PKA treatment, shortening velocity decelerates as sarcomeres traverse from ∼3.10 to ∼3.00 µm. After PKA treatment, sarcomeres shorten a greater distance and exhibit less deceleration during similar force clamps. After sMyBP-C dephosphorylation, sarcomere length traces display a brief recoil (i.e., “bump”) that initiates at ∼3.06 µm during loaded shortening. Interestingly, the timing of the bump shifts with changes in load but manifests at the same sarcomere length. Our results suggest that sMyBP-C and its phosphorylation state regulate sarcomere contraction by a combination of cross-bridge recruitment, modification of cross-bridge cycling kinetics, and alteration of drag forces that originate in the C-zone.

Myofibrillar regulatory mechanisms of stretch activation in mammalian striated muscle

Stretch activation is described as a delayed increase in force after an imposed stretch. This process is essential in the flight muscles of many insects and is also observed, to some degree, in mammalian striated muscles. The mechanistic basis for stretch activation remains uncertain, although it appears to involve cooperative activation of the thin filaments (12, 80). The purpose of this study was to address myofibrillar regulatory mechanisms of stretch activation in mammalian striated muscle. For these studies, permeabilized rat slow-twitch and fast-twitch skeletal muscle fibers were mounted between a force transducer and motor, and a slack-re-stretch maneuver was performed over a range of Ca[superscript 2+] activation levels. Following slack-re-stretch there was a stretch activation process that often resulted in a transient force overshoot (P[subscript TO]), which was quantified relative to steady-state isometric force. P[subscript TO] was highly dependent upon Ca[superscript 2+] activation level, and the relative magnitude of P[subscript TO] was greater in slow-twitch fibers than fast-twitch fibers. In both slow-twitch and fast-twitch fibers, force redevelopment involved a fast, Ca[superscript 2+] activation dependent process (k1) and a slower, less activation dependent process (k2). Interestingly, the two processes converged at low levels of Ca[superscript 2+] activation in both fiber types. P[subscript TO] also contained a relaxation phase, which progressively slowed as Ca[superscript 2+] activation levels increased and was more Ca[superscript 2+] activation dependent in slow-twitch fibers. These results suggest that stretch activation may not be solely regulated by the extent of apparent cooperative activation of force due to a higher relative level of stretch activation in the less cooperative slow-twitch skeletal muscle fiber. Next, we investigated an additional potential molecular mechanism by regulating stretch activation in mammalian striated muscle. Along these lines, our lab has previously observed that PKA-induced phosphorylation of cMyBP-C and cTnI elicited a significant increase in transient force overshoot following slack-re-stretch maneuver in permeabilized cardiac myocytes (29). Interestingly, in slow-twitch skeletal muscle fibers MyBP-C but not ssTnI is phosphorylated by PKA (28). We, thus, took advantage of this variation in substrates phosphorylated by PKA to investigate the effects of PKA-induced phosphorylation of MyBP-C on stretch activation in slow-twitch skeletal muscle fibers. Following PKA treatment of skinned slow-twitch skeletal muscle fibers, the magnitude of P[subscript TO] more than doubled, but this only occurred at low levels of Ca[superscript 2+] activation (i.e., [approximately]25% maximal Ca[superscript 2+] activated force). Also, force redevelopment rates were significantly increased over the entire range of Ca[superscript 2+] activation levels following PKA treatment. In a similar manner, force decay rates showed a tendency of being faster following PKA treatment, however, were only statistically significantly faster at 50% Ca[superscript 2+] activation. Overall, these results are consistent with a model whereby stretch transiently increases the number of cross-bridges made available for force generation and PKA phosphorylation of MyBP-C enhances these stretch activation processes.