An efficient cell transformation method is presented that utilizes droplet electroporation on a microfluidic chip. Two types of green microalgae, a wall-less mutant and a wild type of Chlamydomonas reinhardtii, are used as model cells. The PDMS-glass electroporation chip is simply composed of a flow-focusing microstructure for generating cell-encapsulating droplets and a serpentine channel for better mixing of the content in the droplet, and five pairs of parallel microelectrodes on the glass slide, without involving any expensive electrical equipment. The transformation efficiency via the microfluidic electroporation is shown to be more than three orders of magnitude higher for the wall-less mutant, and more than two orders of magnitude higher for the wild type, which has its cell wall intact, than bulk phase electroporation under identical conditions. Furthermore, the microfluidic transformation is remarkably efficient even at a low DNA/cell ratio, facilitating ways of controlling the transgenic copy number, which is important for the stability of the transgene expression.
This paper presents a microfluidic chip for highly efficient separation of red blood cells (RBCs) from whole blood on the basis of their native magnetic properties. The glass chip was fabricated by photolithography and thermal bonding. It consisted of one inlet and three outlets, and a nickel wire of 69-microm diameter was positioned in the center of a separation channel with 149-microm top width and 73-microm depth by two parallel ridges (about 10 microm high). The two ridges were formed simultaneously during the wet etching of the channels. The nickel wire for generating the magnetic gradient inside the separation channel was introduced from the side of the chip through a guide channel. The external magnetic field was applied by a permanent magnet of 0.3 T placed by the side of the chip and parallel to the main separation channel. The RBCs were separated continuously from the 1:40 (v/v) diluted blood sample at a flow rate in the range 0.12-0.92 microL/min (9-74 mm/min) with the chip, and up to 93.7% of the RBCs were collected in the middle outlet under a flow rate of 0.23 microL/min. The cell sedimentation was alleviated by adjusting the specific density of the supporting media with bovine serum albumin. Quantum dot labeling was introduced for visual fluorescence tracking of the separation process. The uneven distribution phenomenon of the blood cells around the nickel wire was reported and discussed.
Ferromagnetic Fe3O4-doped MgB2 bulks were first fabricated in this work by the hot pressing method. It was found that Fe3O4 does not react with Mg or B during the fabrication process. Peak Jc values of the 5 wt% Fe3O4-doped MgB2 are higher than 106 A cm−2 in the temperature range 5–30 K. Especially at 30 K, the peak Jc is 1.02 × 106 A cm−2 for the 5 wt% Fe3O4-doped MgB2, the highest values at 30 K found in the literature, and about seven times that of the 5 wt% SiC-doped MgB2 sample. The drop in Jc with increasing field for the Fe3O4-doped MgB2 is significantly slower than that of the SiC-doped MgB2 at 30 K. These results indicate that the Fe3O4-doped MgB2 is a potential superconductor to be used at temperatures greater than 25 K which is a critical temperature for large-scale practical applications.
Bead injection in a lab-on-valve (LOV) system was adopted for DNA purification via micro solid-phase extraction (SPE) with a renewable silica microcolumn packed in a channel of the LOV unit. The complex matrix components in human whole blood, including proteins, were well eliminated by choosing properly the sample loading and elution media. The DNA purification process was monitored on-line by using laser-induced fluorescence in a demountable side part of the LOV unit incorporating optical fibers. The practical applicability of the entire system was demonstrated by separation/purification of lambda-DNA in a simulated matrix and human blood genetic DNA by performing SPE, in situ monitoring of the purified products, and postcolumn PCR amplification. When DNAs in a simulated matrix (10.0 ng microl-1 lambda-DNA, 50 ng microl-1 bovine serum albumin, 1.0% Triton X-100) were processed in the present system and laser-induced fluorescence was monitored at 610 nm, an overall extraction/collection efficiency of 70% was achieved by employing identical sample loading and an elution flow rate of 0.5 microl s-1, along with a precision of 3.8% relative standard deviation. DNA separation and purification from human whole-blood samples were performed under similar conditions.
We report the phase evolution, microstructure, and critical current density (Jc) of the SiC doped MgB2 superconductors. In our study, all samples were fabricated by hot pressing with a heating rate of 200 °C min−1. The results show that the reaction of 2Mg+SiC = Mg2Si+C can occur at about 500 °C, about 50 °C lower than the formation temperature of MgB2. On the other hand, according to the experimental results and thermodynamic calculations, boron (B) does not react with SiC to form B4C and Si, neither does MgB2 react with SiC to form Mg2Si and B4C. The MgB2 phase was formed via both a solid–solid reaction (during the heating process between about 500 and 650 °C) and a liquid–solid reaction (>650 °C) after the melting of Mg, and the two reactions resulted in differences in the grain size of the MgB2. From scanning electron microscopy, the Mg2Si particles are homogeneously distributed within the MgB2 matrix, with particle sizes ranging from 35 to 230 nm. From the perspective of superconductivity, the C substitution results in strong electron scattering centers in the MgB2 structure. It reduces the electron mean free path and thus may significantly enhance the magnetic Jc. The peak Jc of the 5 wt% SiC doped MgB2 reaches above 106 A cm−2 at 5 K, and decreases slowly with increasing field, remaining high, above 105 A cm−2, at 7 T.
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