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In cells, motor proteins do not work alone but rather move in ensemble when they support several cellular functions including mitosis and beating of flagellum. Until now, collective transportation by motors was poorly understood due to the experimental difficulty in controlling the number and arrangement of motors, which are considered to affect the transportation. In this report, we propose a novel...
In this study, we developed a microfluidic platform for a three-dimensional tissue model with a perfusable capillary network, which will allow, for the first time, a perfusion-culture in a tissue model with a high cell density. Our group previously reported that a spheroid of lung fibroblasts induced angiogenic sprouts from microchannels [1]. In this study, we successfully connected angiogenic sprouts...
We propose a nano-patterning method to control the number of kinesin motors involving in a collective transport of a single microtubule (MT) filament. We fabricated Au nano-pillar array on a thermally oxidized silicon substrate. SiO2 surface was selectively coated with poly(ethylene glycol) (PEG) self-assembled monolayer (SAM) and was rendered inert to protein adsorption. Streptavidin was immobilized...
We developed Linear Zero Mode Waveguides (LZMWs) for simultaneous observation of kinesin displacement and ATP hydrolysis cycle under higher concentration of fluorescently labeled ATP than that used in conventional total internal reflection microscopy (TIRFM). Single-molecule fluorescent microscopy is a promising method to analyze correlation between the mechanical displacement and hydrolysis of ATP...
For the analysis of kinesin motility, we demonstrate an observation system to simultaneously visualize kinesin and ATP molecules at the single molecule level using Linear Zero-Mode Waveguides (LZMWs). LZMWs allow fluorescent imaging at up to micromolar concentration by confining the excitation volume, which is high enough for studying enzymatic reactions such as ATP hydrolysis. LZMWs were fabricated...
This paper describes a method for controlling the velocity of gliding microtubules by light irradiance on a gold-patterned surface. The light was used to control the temperature of the buffer solution by heat transfer from the thin gold film. Microtubule velocity increased approximately 1.8-fold from the initial velocity at the 13.5 W/cm2 on the gold-coated area. The proposed method also allows cyclical...
We propose a method to control gliding directions of kinesin-propelled microtubules (MTs) corresponding to their flexural rigidity (EI). We prepared two kinds of MTs having different EI and their trajectories within an electric field were clearly separated. Therefore, this study demonstrated the EI-altered MTs can be workhorses to sort/concentrate various combinations of target molecules loaded on...
In this paper, we present a poly-dimethylsiloxane (PDMS) microfluidic device to create a vascular network for a long-term spheroid culture, of which network and spheroid are consist of human umbilical vein endothelial cells (HUVEC) and normal human lung fibroblasts (LF), respectively. Following device design, fabrication, and fundamental evaluation of HUVEC sprouting conditions, we visualized that...
Microtubule gliding on a kinesin-coated surface may be applied as a nanoactuator to manipulate nanoscale materials in a microfluidic environment. However, controlling the velocity and direction of microtubule gliding has been a major challenge for bioengineering. To develop a novel method of molecular manipulation, we propose a technique to control the velocity of gliding microtubules by light irradiance,...
In this paper, we show a reconstruction of kinesin driven transport system in three dimensionally (3D) enclosed channels whose scale is similar to axons. Our experimental method enabled successful motility of a large number of kinesin-labeled Q-dots on microtubules (MTs) in enclosed channels. To control the direction of kinesin motility, we prepared a polarity-defined MT array in channels. Due to...
We developed a system to visualize molecular bindings as colocalization of two Q-dots carried by motor proteins on a microtubule (MT) array with predefined polarities in nanotracks. The MT array was prepared utilizing gliding of MTs in micro/nano structures and a dissociation method of retrogressing MTs. Pairs of molecules for cargo transport was designed and evaluated experimentally. One of molecular...
Piezoelectric shear strain was measured for c-axis oriented epitaxial Pb(Zr,Ti)O3 (PZT) thin films. The PZT films, with a composition near the morphotropic phase boundary (MPB), were epitaxially grown on (001)MgO substrates and then microfabricated into a rectangular shape. Lateral electrodes were deposited on both sides of the PZT films to apply an external electric field perpendicular to the polarization...
In this study, we fabricate multilayer ceramic capacitors (MLCCs) composed of BaTiO3 (BT) dielectric layers with Pt internal electrodes by radio frequency magnetron sputtering. We deposited BT layers with thickness of approximately 300 nm on Pt/Ti-coated SiO2/Si substrates through a movable shadow mask. The isolated internal Pt electrodes are prepared by sliding the shadow mask. Multilayered BT thin...
In this study, we have developed Ti-based piezoelectric MEMS scanner mirrors driven by PZT thin films. The 2 µm-thick PZT thin films were directly deposited on 50 µm-thick Ti substrates by rf-magnetron sputtering. The Ti substrates were preliminary microfabricated into the shape of the scanner mirror by wet-etching, therefore we could fabricate piezoelectric micro-actuators without conventional photolithographic...
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