35904594
Targeted cell immobilization by ultrasound microbeam.
Various techniques exerting mechanical stress on cells have been developed to investigate cellular responses to externally controlled stimuli. Fundamental mechanotransduction processes about how applied physical forces are converted into biochemical signals have often been examined by transmitting such forces through cells and probing its pathway at cellular levels. In fact, many cellular biomechanics studies have been performed by trapping (or immobilizing) individual cells, either attached to solid substrates or suspended in liquid media. In that context, we demonstrated two-dimensional acoustic trapping, where a lipid droplet of 125 µm in diameter was directed transversely toward the focus (or the trap center) similar to that of optical tweezers. Under the influence of restoring forces created by a 30 MHz focused ultrasound beam, the trapped droplet behaved as if tethered to the focus by a linear spring. In order to apply this method to cellular manipulation in the Mie regime (cell diameter > wavelength), the availability of sound beams with its beamwidth approaching cell size is crucial. This can only be achieved at a frequency higher than 100 MHz. We define ultrasound beams in the frequency range from 100 MHz to a few GHz as ultrasound microbeams because the lateral beamwidth at the focus would be in the micron range. Hence a zinc oxide (ZnO) transducer that was designed and fabricated to transmit a 200 MHz focused sound beam was employed to immobilize a 10 µm human leukemia cell (K-562) within the trap. The cell was laterally displaced with respect to the trap center by mechanically translating the transducer over the focal plane. Both lateral displacement and position trajectory of the trapped cell were probed in a two-dimensional space, indicating that the retracting motion of these cells was similar to that of the lipid droplets at 30 MHz. The potential of this tool for studying cellular adhesion between white blood cells and endothelial cells was discussed, suggesting its capability as a single cell manipulator.
39975301
Effect of ultrasonic attenuation on the feasibility of acoustic tweezers.
A modified mathematical formulation for the calculation of axial radiation force was developed to incorporate the effect of ultrasonic attenuation. Axial forces, Fresnel coefficients, average internal attenuation factors and effective internal reflection coefficients were calculated. Thermal and mechanical indices were also computed to address the safety issues in the implementation of acoustic tweezers and were found to be negligible. The results show that the overall distribution of axial forces is barely affected by attenuation. Furthermore, it is found that attenuation actually works against the scattering force and may therefore reinforce the axial trapping force. For a particle size of 180 microm, the maximum trapping force increases from 29.8 x 10(-11) N to 30.3 x 10(-11) N by 1.7% when attenuation is included. In light of these results, it appears that acoustic tweezers may still be feasible beyond the focal point even under the influence of attenuation.
1587472
Calibration of sound forces in acoustic traps
A two-dimensional or transverse acoustic trapping and its capability to noninvasively manipulate micrometersized particles with focused sound beams were experimentally demonstrated in our previous work. To apply this technique, as in optical tweezers, for studying mechanical properties of and interactions among biological particles such as cells, the trapping forces must be calibrated against known forces, i.e., viscous drag forces exerted by fluid flows. The trapping forces and the trap stiffness were measured under various conditions and the results were reported in this paper. In the current experimental arrangement, because the trapped particles were positioned against an acoustically transparent mylar membrane, the ultrasound beam intensity distribution near the membrane must be carefully considered. The total intensity field (the sum of incident and scattering intensity fields) around the droplet was thus computed by finite element analysis (FEA) with the membrane included, and it was then used in the ray acoustics model to calculate the trapping forces. The membrane effect on trapping forces was discussed by comparing effective beam widths with and without the membrane. The FEA results found that the broader beam width, caused by the scattered beams from the neighboring membrane and the droplet, resulted in the lower intensity, or smaller force, on the droplet. The experimental results showed that the measured forces were as high as 64 nN. The trap stiffness, approximated as a linear spring, was estimated by linear regressions and found to be 1.3 to 4.4 nN/μm, which was on a larger scale than that of optical trapping estimated for red blood cells, a few tenths of piconewtons/nanometer. The experimental and theoretical results were in good agreement.
21739268
Transverse acoustic trapping using a gaussian focused ultrasound.
The optical tweezer has become a popular device to manipulate particles in nanometer scales and to study the underlying principles of many cellular or molecular interactions. Theoretical analysis was previously carried out at the authors' laboratory, to show that similar acoustic trapping of microparticles may be possible with a single beam ultrasound. This article experimentally presents the transverse trapping of 125 microm lipid droplets under an acoustically transparent mylar film, which is an intermediate step toward achieving acoustic tweezers in three-dimension. Despite the lack of axial trapping capability in the current experimental arrangement, it was found that a 30 MHz focused beam could be used to laterally direct the droplets toward the focus. The spatial range within which acoustic traps may guide droplet motion was in the range of hundreds of micrometers, much greater than that of optical traps. This suggests that this acoustic device may offer an alternative for manipulating microparticles in a wider spatial range.
13196701
A biomimetic mass-flow transducer utilizing all-optofluidic generation of self-digitized, pulse code-modulated optical pulse trains.
We present a new mass-flow transducer producing responses in the form of optical pulse trains that are encoded with information on the strength and position of the stimulus. We implemented the self-digitization and encoding capabilities all-optofluidically, without involving external electronics, by integrating one optical fiber cantilever with multiple polymer optical waveguides on a microfluidic platform. The transducer can also be configured to respond only to transitional stimuli. These features closely mimic the rate-coding, action potential labeling, and rapid adaptation processes observed in biological mechanoreceptors and allow multiple transducers to transmit signals over a single, shared channel. We fabricated the transducer using polymer-based soft-lithography techniques. Its characterization confirmed the stimulus strength-dependent generation of optical pulses and the feasibility of multiplexing 2(n-1) to 2(n) transducers using n waveguides.
4429405
Regulation of cell movement is mediated by stretch-activated calcium channels
Intracellular calcium regulates many of the molecular processes that are essential for cell movement. It is required for the production of actomyosin-based contractile forces, the regulation of the structure and dynamics of the actin cytoskeleton,, and the formation and disassembly of cell–substratum adhesions,. Calcium also serves as a second messenger in many biochemical signal-transduction pathways. However, despite the pivotal role of calcium in motile processes, it is not clear how calcium regulates overall cell movement. Here we show that transient increases in intracellular calcium, [Ca2+]i, during the locomotion of fish epithelial keratocytes, occur more frequently in cells that become temporarily ‘stuck’ to the substratum or when subjected to mechanical stretching. We find that calcium transients arise from the activation of stretch-activated calcium channels, which triggers an influx of extracellular calcium. In addition, the subsequent increase in [Ca2+]i is involved in detachment of the rear cell margin. Thus, we have defined a mechanism by which cells can detect and transduce mechanical forces into biochemical signals that can modulate locomotion.
6921183
The effect of storage conditions on the permeability of porcine buccal mucosa
The impact of storage conditions on the permeability of porcine buccal mucosa to [3H]water and [14C]mannitol was assessed. The fresh porcine buccal tissue (fresh tissue) was obtained by utilizing pig heads within 24 hours of slaughter. The stored and frozen porcine buccal tissues (stored tissue and frozen tissue) were obtained after the storage of the tissue intact in the pig heads at 4 degrees C or -20 degrees C, respectively, for 24 h. The results demonstrated that the barrier properties of the porcine buccal mucosa were maintained with regard to [3H]water permeability when stored at 4 degrees C for 24 h. However, freezing the tissue resulted in tissue damage illustrated by a significant increase in [3H]water permeability. [14C]Mannitol does not appear to be a suitable model solute to assess the ex vivo permeability of porcine buccal mucosa due to its extremely low permeability.
12699756
Chicoric acid: chemistry, distribution, and production
Though chicoric acid was first identified in 1958, it was largely ignored until recent popular media coverage cited potential health beneficial properties from consuming food and dietary supplements containing this compound. To date, plants from at least 63 genera and species have been found to contain chicoric acid, and while the compound is used as a processing quality indicator, it may also have useful health benefits. This review of chicoric acid summarizes research findings and highlights gaps in research knowledge for investigators, industry stakeholders, and consumers alike. Additionally, chicoric acid identification, and quantification methods, biosynthesis, processing improvements to increase chicoric acid retention, and potential areas for future research are discussed.
7202726
Improved shim method based on the minimization of the maximum off-resonance frequency for balanced steady-state free precession (bSSFP).
In this work, a shim method that minimizes the maximum off-resonance frequency (min-max shim) in balanced steady-state free precession (bSSFP) is tested for brain imaging at 3T with constant and linear shim terms. The method demonstrates improvement of spatial coverage and banding artifact reduction over standard least-squares shimming. In addition, a new method (modified min-max shim) is introduced. This method reduces boundary band regions where the artifact is inevitable due to the excessive off-resonance frequency distribution. In comparison to standard least-squares shimming, the min-max based shim method either eliminates or reduces the size of banding artifacts. The method can be used to increase the signal-to-noise ratio (SNR) in bSSFP imaging or to increase the functional contrast in bSSFP functional MRI (fMRI) by allowing a longer usable repetition time (TR).