Continuous-flow polymerase chain reaction microfluidics by using spiral capillary channel embedded on copper

This paper presents a novel method for performing polymerase chain reaction (PCR) amplification by using spiral channel fabricated on copper where a transparent polytetrafluoroethylene ( PTFE) capillary tube was embedded. The channel with 25 PCR cycles was gradually developed in a spiral manner from inner to outer. The durations of PCR mixture at the denaturation, annealing and extension zones were gradually lengthened at a given flow rate, which may benefit continuous-flow PCR amplification as the synthesis ability of the Taq polymerase enzyme usually weakens with PCR time. Successful continuous-flow amplification of DNA fragments has been demonstrated. The PCR products of 249, 500 and 982 bp fragments could be obviously observed when the flow rates of PCR mixture were 7.5, 7.5 and 3.0 mm s(-1), respectively, and the required amplification times were about 25, 25, and 62 min, respectively. Besides, the successful segmented-flow PCR of three samples ( 249, 500 and 982 bp) has also been reported, which demonstrates the present continuous-flow PCR microfluidics can be developed for high-throughput genetic analysis.

Transient flow patterns and bubble slug lengths in parallel microchannels with oxygen gas bubbles produced by catalytic chemical reactions

Transient flow patterns and bubble slug lengths were investigated with oxygen gas (O-2) bubbles produced by catalytic chemical reactions using a high speed camera bonded with a microscope. The microreactor consists of an inlet liquid plenum, nine parallel rectangular microchannels followed by a micronozzle, using the MEMS fabrication technique. The etched surface was deposited by the thin platinum film, which is acted as the catalyst. Experiments were performed with the inlet mass concentration of the hydrogen peroxide from 50% to 90% and the pressure drop across the silicon chip from 2.5 to 20.0 kPa. The silicon chip is directly exposed in the environment thus the heat released via the catalytic chemical reactions is dissipated into the environment and the experiment was performed at the room temperature level. It is found that the two-phase flow with the catalytic chemical reactions display the cyclic behavior. A full cycle consists of a short fresh liquid refilling stage, a liquid decomposition stage followed by the bubble slug flow stage. At the beginning of the bubble slug flow stage, the liquid slug number reaches maximum, while at the end of the bubble slug flow stage the liquid slugs are quickly flushed out of the microchannels. Two or three large bubbles are observed in the inlet liquid plenum, affecting the two-phase distributions in microchannels. The bubble slug lengths, cycle periods as well as the mass flow rates are analyzed with different mass concentrations of hydrogen peroxide and pressure drops. The bubble slug length is helpful for the selection of the future microreactor length ensuring the complete hydrogen peroxide decomposition. Future studies on the temperature effect on the transient two-phase flow with chemical reactions are recommended.

Oscillation flow induced by underwater supersonic gas jets

This paper describes an experimental study on the oscillation flow characteristics of submerged supersonic gas jets issued from Laval nozzles. The flow pattern during the jet development and the jet expansion feedback phenomenon are studied using a high-speed camera and a pressure measurement system. The experimental results indicate that along the downstream distance, the jet has three flow regimes: (1) momentum jet; (2) buoyant jet; (3) plume. In the region near the nozzle exit a so-called bulge phenomenon is found. Bulging of the jet occurs many times before the more violent jet expansion feedback occurs. During the feedback process, the jet diameter can become several times that of the original one depending on the jet Mach number. The frequencies of the jet bulging and the jet expansion feedback are measured.

Effects of forced solution flow on lysozyme crystal growth

In order to study quantitatively the effects of forced solution on crystal growth, we designed a new set of experimental equipment, in particular, a microchannel mixer was used as crystallization container so that the consumption of protein samples was much reduced and thus an exact syringe pump could be used for precise control of the flow rates. Since the mixer’s section was designed to be rectangular, the solution velocity in its center was steady and constant, and thus repeatable experiments were facilitated. Experimental results showed that the effects of forced solution on protein crystal growth were different under different levels of supersaturation, and new results were obtained for cases of high supersaturation. When the supersaturation is σ = 2.3, with increasing flow rates the growth rates of the lysozyme crystal’s (110) face hardly change when the flow rates are lower than 1300 μm/s, and decrease quickly afterwards. When the flow rate reaches 2000 μm/s, the crystal nearly ceases to grow. When the supersaturation is σ = 2.7, with increasing flow rates the (110) face growth rates increase at the beginning then reach the maximum values at 1700 μm/s – 1900 μm/s and decrease afterwards, approaching zero or so when the flow rate reaches 12000 μm/s. The higher the supersaturation, the larger the flow rate at which the crystal ceases to grow. © 2010 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim

Quasi-3D Cytoskeletal Dynamics of Osteocytes under Fluid Flow

Osteocytes respond to dynamic fluid shear loading by activating various biochemical pathways, mediating a dynamic process of bone formation and resorption. Whole-cell deformation and regional deformation of the cytoskeleton may be able to directly regulate this process. Attempts to image cellular deformation by conventional microscopy techniques have been hindered by low temporal or spatial resolution. In this study, we developed a quasi-three-dimensional microscopy technique that enabled us to simultaneously visualize an osteocyte's traditional bottom-view profile and a side-view profile at high temporal resolution. Quantitative analysis of the plasma membrane and either the intracellular actin or microtubule (MT) cytoskeletal networks provided characterization of their deformations over time. Although no volumetric dilatation of the whole cell was observed under flow, both the actin and MT networks experienced primarily tensile strains in all measured strain components. Regional heterogeneity in the strain field of normal strains was observed in the actin networks, especially in the leading edge to flow, but not in the MT networks. In contrast, side-view shear strains exhibited similar subcellular distribution patterns in both networks. Disruption of MT networks caused actin normal strains to decrease, whereas actin disruption had little effect on the MT network strains, highlighting the networks' mechanical interactions in osteocytes.

Investigations of phase inversion and frictional pressure gradients in upward and downward oil–water flow in vertical pipes

The present study has attempted to investigate phase inversion and frictional pressure gradients during simultaneous vertical flow of oil and water two-phase through upward and downward pipes. The liquids selected were white oil (44 mPa s viscosity and 860 kg/m3 density) and water. The measurements were made for phase velocities varying from 0 to 1.24 m/s for water and from 0 to 1.87 m/s for oil, respectively. Experiments were carried either by keeping the mixture velocity constant and increasing the dispersed phase fraction or by keeping the continuous phase superficial velocity constant and increasing the dispersed phase superficial velocity. From the experimental results, it is shown that the frictional pressure gradient reaches to its lower value at the phase inversion point in this work. The points of phase inversion are always close to an input oil fraction of 0.8 for upward flow and of 0.75 for downward flow, respectively. A few models published in the literature are used to predict the phase inversion point and to compare the results with available experimental data. Suitable methods are suggested to predict the critical oil holdup at phase inversion based on the different viscosity ratio ranges. Furthermore, the frictional pressure gradient is analyzed with several suitable theoretical models according to the existing flow patterns. The analysis reveals that both the theoretical curves and the experimental data exhibit the same trend and the overall agreement of predicted values with experimental data is good, especially for a high oil fraction.

Multi-mode Vortex-induced Vibration of Slender Cable Experiencing Shear Flow

The dynamic characteristics of slender cable often present serried modes with low frequencies due to large structure flexibility resulted from high aspect ratio (ratio of length to diameter of cable), while the flow velocity distributes non-uniformly along the cable span actually in practical engineering. Therefore, the prediction of the vertex-induce vibration of slender cable suffered from multi-mode and high-mode motions becomes a challenging problem. In this paper a prediction approach based on modal energy is developed to deal with multi-mode lock-in. Then it is applied to the modified wake-oscillator model to predict the VIV displacement and stress responses of cable in non-uniform flow field. At last, illustrative examples are given of which the VIV response of flexible cable in nonlinear shear flow field is analyzed. The effects of flow velocity on VIV are explored. Our results show that both displacement and stress responses become larger as the flow velocity increasing; especially higher stress response companied with higher frequency vibration should be paid enough attention in practical design of SFT because of its remarkable influence on structure fatigue life.

Measurement of neutron flow with the external target experimental device on CSR

With the construction of the neutron detection wall at the external target position on Heavy Ion Research Facility in Lanzhou-Cooling Storage Ring (HIRFL-CSR), it will be possible to detect high energy neutron. A BUU model is applied to simulate the flow in both symmetric (Ni+Ni, Pb+Pb) and asymmetric(Pb+Ni) systems. It is shown that at above several hundreds MeV/u, the flow signals are very obvious and depend clearly on the centrality of the collisions. Based on the products in the forward angle less than 20 degrees, the simulation also reveals that the determination of the reaction plane and the selection of the impact parameter, both of which are essential in the flow measurement, are well implemented. The double event and its influence on the determination of the neutron flow are also simulated.

Protective effects of melatonin against low- and high-LET irradiation

To investigate the protective effects of melatonin against high-LET ionizing radiation, V79 Chinese hamster cells were irradiated with 100 keV/mu m carbon beam. Parallel experiments were performed with 200 kV X-rays. To avoid the impact from extra solvents, melatonin was dissolved directly in culture medium. Cells were cultured in melatonin medium for 1 hr before irradiation. Cell inactivation was measured with conventional colony forming assay, medium containing 6-thioguanine was used for the selection of mutants at hprt locus, and the cell cycle was monitored by flow cytometry. Both carbon beam and X-rays induced cell inactivation, hprt gene mutation and cell cycle G2 block dose-dependently. But carbon beam showed stronger effects as indicated by all three endpoints and the relative biological effectiveness (RBE) was 3.5 for cell killing (at 10% survival level) and 2.9 for mutation induction (at 5 x 10(-5) mutants/ cell level). Melatonin showed protective effects against ionizing radiation in a dose-dependent manner. In terms of cell killing, melatonin only increased the survival level of those samples exposed to 8Gy or larger of X-rays or 6 Gy or larger of carbon beam. In the induction of hprt mutation and G2 block, melatonin reduced such effects induced by carbon beam but not by X-rays. The results suggest that melatonin reduces the direct interaction of particles with cells rather than an indirect interaction. Further studies are required to disclose the underlying mechanisms.

High-efficiency transfer and expression of AdCMV-p53 in human hepatocellular carcinoma cells induced by low-dose carbon-ion radiation

Objective To investigate whether the irradiation with C-beam could enhance adenovirus-mediated transfer and expression of p53 in human hepatocellular carcinoma. Materials and methods HepG2 cells were exposed to C-beam or gamma-ray and then infected with replicationdeficient adenovirus recombinant vectors containing human wild-type p53 or green fluorescent protein, respectively. The transfer efficiency and expression level of the exogenous gene were detected by flow cytometric analysis. Cell survival fraction was detected by clonogenic assay. Results The transfer frequency in C-beam or gamma-irradiated groups increased by 50-83% and 5.7-38.0% compared with the control, respectively (P < 0.05). Compared with C-beam alone, p53 alone, and gamma-ray with p53, the percentages of p53 positive cells for 1 Gy C-beam with p53 increased by 56.0-72.0%, 63.5-82.0%, and 31.3-72.5% on first and third day after the treatments, respectively (P < 0.05). The survival fractions for the 2Gy C-bearn and AdCMV-p53 infection groups decreased to similar to 2%. Conclusion C-beam irradiation could significantly promote AdCMV-green fluorescent protein transfer and expression of p53.

TRITON-(HE)-H-3 RELATIVE AND DIFFERENTIAL FLOWS AND THE HIGH DENSITY BEHAVIOR OF NUCLEAR SYMMETRY ENERGY

Using a transport model coupled with a phase-space coalescence after-burner we study the triton-He-3 relative and differential transverse flows in semi-central Sn-132 + Sn-124 reactions at a beam energy of 400 MeV/nucleon. We find that the triton-He-3 pairs carry interesting information about the density dependence of the nuclear symmetry energy. The t-He-3 relative flow can be used as a particularly powerful probe of the high-density behavior of the nuclear symmetry energy.