Journal: PLOS One

Article Title: Gene electrotransfer with flow-through microchannel and lower alternating voltage generated induced pluripotent cells from human lymphoblastoid cell lines

doi: 10.1371/journal.pone.0333491

Figure Lengend Snippet: (a) A schematic drawing of a microchannel chip for gene electrotransfer. Cells and plasmid vector suspension were introduced into single-lined microchannel via pipette tip and pneumatic pressure. An alternating biphasic wave, either sinusoidal or triangular, was applied across the electrodes, forming an electric field around them that facilitated plasmid vector transfer into cells. (b) Fabricated microchannel chip with inlet and outlet biopsy punches of 1- and 3-mm diameter, respectively. An unused branch channel for droplet formation was also visible . The scale bar is 200 µm. (c) Experimental setup for flow generation and voltage application. A pneumatic pump (Flow EZ 7000 mbar, Fluigent) was attached to the pipette tip at the inlet. The electric power source consisted of a waveform generator (33500B, Agilent) connected to a bipolar amplifier (BA4825, NF Co.). A 1 kΩ resistor was connected to the electrodes, and an oscilloscope (PicoScope 6404E, P2056, Pico Technology) monitored the electrode voltage. The voltage amplitude decreased as the solution contacts the electrodes, and amplitude fluctuations indicated cell passage, which helped to monitor channel activity.

Article Snippet: The flow within the microchannel was visualized using an inverted microscope (Eclipse Ti2-U, Nikon) and a high-speed camera (HAS-D71G, Ditect).

Techniques: Electrotransfer, Plasmid Preparation, Suspension, Transferring, Activity Assay

Journal: PLOS One

Article Title: Gene electrotransfer with flow-through microchannel and lower alternating voltage generated induced pluripotent cells from human lymphoblastoid cell lines

doi: 10.1371/journal.pone.0333491

Figure Lengend Snippet: (a) Intensity of electric field in the microchannel without cells. (b) Intensity of electric field when a cell is centered in the channel between electrodes at 1 ms. (c) Transmembrane potential distribution on cell membrane at 1 ms based on asymptotic Smoluchowski equations. (d) Distribution of pore density on the plasma membrane. (e) Distribution of pore density on plasma membrane circumference at various application voltages at a time of 1 ms. (f) Pore density at q = 0, and 1 ms in various applications.

Article Snippet: The flow within the microchannel was visualized using an inverted microscope (Eclipse Ti2-U, Nikon) and a high-speed camera (HAS-D71G, Ditect).

Techniques: Membrane, Clinical Proteomics

Journal: PLOS One

Article Title: Gene electrotransfer with flow-through microchannel and lower alternating voltage generated induced pluripotent cells from human lymphoblastoid cell lines

doi: 10.1371/journal.pone.0333491

Figure Lengend Snippet: (a) Images of LCLs with bright field (BF), the fluorescence of Hoechst 33342 (blue), PI (red), and GFP expression (green). Bright-field images and fluorescent-filtered images with the filters of FITC, mCherry, and DAPI are taken, respectively. Positions of cells were slightly moved while taking with each fluorescent filter because LCLs are floating cells. Scale bar = 200 µm. (b) Microscopic images of microchannel around electrode pair. White arrows show the location of electrode elution. Bubbling and elution of electrodes are sometimes observed above 12.5 V application. Scale bar = 100 µm. (c) Transfection efficiency and viability of LCLs as a parameter of application voltages for sinusoidal wave and triangle wave. Significant differences between 0V application and other voltages are indicated by #. Significant differences between triangle and sinusoidal waves are depicted by *.

Article Snippet: The flow within the microchannel was visualized using an inverted microscope (Eclipse Ti2-U, Nikon) and a high-speed camera (HAS-D71G, Ditect).

Techniques: Fluorescence, Expressing, Transfection