Kaoru Uesugi

Nano- and micro-structures can be realized by using photofabrication, crystal growth, chemical deposition, etching and other methods. This variety of fabrication methods encourages the development of water-repellent surfaces from the viewpoint of biomimetics. Water striders have attracted interest in biomimetics because their water-repellency property makes it possible for them to live on water surfaces. Micro-hairs, which are present on the surface of the water strider’s legs, can maintain an air layer between the water surface and the legs, providing the repellency property. While there have been various studies which have considered a physical model of the water strider’s water-repellency property, no model has taken directly measured water strider leg-rowing force into account. Therefore, in this study, we proposed a physical model of a water strider using Laplace pressure, and then we considered directly measured leg-rowing force to verify the model. First, we considered the relationship between water pressure around the micro-hairs and the intersection of the surface of the micro-hairs and the water surface using our proposed model. We found that when the micro-hairs were in contact with the water surface and the air layer was maintained, the maximum Laplace pressure P_max was 35.2 kPa. This meant that if Laplace pressure exceeded 35.2 kPa, the water pressure caused by the rowing motion of the strider legs pushed water into the space between micro-hairs. Additionally, we calculated maximum water pressure P_L which was loaded around the surface of the water strider's leg by rowing of the leg. P_L was 546 Pa, and this value was significantly smaller than P_max. This meant that the water pressure did not push water into the space between the leg micro-hairs and the water strider’s legs maintained their water repellency when moving.

A ring pump (RP) is a useful tool for microchannels and automated cell culturing. We have been developing RPs (a full-press ring pump, FRP; and a mid-press ring pump, MRP). However, damage to cells which were sent by the RP and the MRP was not investigated, and no other studies have compared the damage to cells between RPs and peristaltic pumps (PPs). Therefore, first, we evaluated the damage to cells that were sent by a small size FRP (s-FRP) and small size MRPs (s-MRPs; gap = 25 or 50 μm, respectively). “Small size” means that the s-FRP and the s-MRPs are suitable for microchannel-scale applications. The survival rate of cells sent by the s-MRPs was higher than those sent by the s-FRP, and less damage caused by the former. Second, we compared the survival rate of cells that were sent by a large size FRP (l-FRP), a large size MRP (l-MRP) (gap = 50 μm) and a PP. “Large size” means that the l-FRP and the l-MRP are suitable for automated cell culture system applications. We could not confirm any differences among the cell survival rates. On the other hand, when cells suspended in Dulbecco’s phosphate-buffered saline solution were circulated with the l-MRP (gap = 50 μm) and the PP, we confirmed a difference in cell survival rate, and less damage caused by the former.

In this report, we propose a micro vacuum chuck (MVC) which can connect three-dimensional (3D) tissues to a tensile test system by vacuum pressure. Because the MVC fixes the 3D tissue by vacuum pressure generated on multiple vacuum holes, it is expected that the MVC can fix 3D tissue to the system easily and mitigate the damage which can happen by handling during fixing. In order to decide optimum conditions for the size of the vacuum holes and the vacuum pressure, various sized vacuum holes and vacuum pressures were applied to a normal human cardiac fibroblast 3D tissue. From the results, we confirmed that a square shape with 100 µm sides was better for fixing the 3D tissue. Then we mounted our developed MVCs on a specially developed tensile test system and measured the bio-mechanical property (beating force) of cardiac 3D tissue which was constructed of human induced pluripotent stem cell-derived cardiomyocytes (hiPS-CM); the 3D tissue had been assembled by the layer-by-layer (LbL) method. We measured the beating force of the cardiac 3D tissue and confirmed the measured force followed the Frank-Starling relationship. This indicates that the beating property of cardiac 3D tissue obtained by the LbL method was close to that of native cardiac tissue.

Bio-actuators that use insect muscular tissue have attracted attention from researchers worldwide because of their small size, self-motive property, self-repairer ability, robustness, and the need for less environment management than mammalian cells. To demonstrate the potential of insect muscular tissue for use as bio-actuators, three types of these robots, a pillar actuator, a walker, and a twizzer, have been designed and fabricated. However, a model of an insect muscular tissue-powered swimming robot that is able to float and swim in a solution has not yet been reported. Therefore, in this paper, we present a prototype of an insect muscular tissue-powered autonomous micro swimming robot that operates at room temperature and requires no temperature and pH maintenance. To design a practical robot body that is capable of swimming by using the force of the insect dorsal vessel (DV), we first measured the contraction force of the DV. Then, the body of the swimming robot was designed, and the design was confirmed by a simulation that used the condition of measured contraction force. After that, we fabricated the robot body using polydimethylpolysiloxane (PDMS). The PDMS body was obtained from a mold that was fabricated by a stereo lithography method. Finally, we carefully attached the DV to the PDMS body to complete the assembly of the swimming robot. As a result, we confirmed the micro swimming robot swam autonomously at an average velocity of 11.7 μm/s using spontaneous contractions of the complete insect DV tissue. These results demonstrated that the insect DV has potential for use as a bio-actuator for floating and swimming in solution.

In single cell analysis, transport of foreign substances into a cell is an important technique. In particular, for accurate analysis, a method to transport a small amount (pico-liter order) of substance into the cell without leakage while retaining the cell shape is essential. Because the fusion of the cell and the giant liposome is a closed system to the outside, it may be possible to transport a precise, small amount of substances into the cell. Additionally, there is no possibility that a leaked substance would affect other systems. To develop the liposome-cell transportation system, knowledge about the behavior of substances in the liposome and the cell is important. However, only a few studies have observed the substance transport between a liposome and a cell. Here, we report observation of small amount of substance transport into a single C2C12 cell by using a giant liposome. Substance transport occurred by electrofusion between the cell and the giant liposome containing the substance, which is a closed system. First, to observe the electrofusion and substance transport from the moment of voltage application, we fabricated a microfluidic device equipped with electrodes. We introduced suspensions of cells and liposomes into the microfluidic device and applied alternating current (AC) and direct current (DC) voltages for electrofusion. We observed a small amount (22.4 ± 0.1%, 10.3 ± 0.4% and 9.1 ± 0.1%) of fluorescent substance (Calcein) contained in the liposomes was transported into the cell without leakage outside the cell, and we obtained the diffusion coefficient of Calcein in the cell as 137 ± 18 μm²/s. We anticipate that this system and the knowledge acquired will contribute to future realization of more accurate single cell analysis in a wide range of fields.

We report a new analytical model of the MEMS-based thermal property sensor for samples which are difficult to handle and susceptible to damage by thermal stimulus, such as living cells. Many sensor designs had been reported for thermal property measurements, but only a few of them have considered the analytical model of the single-sided measurement in which a measurement sample is placed on the sensor substrate. Even in the few designs that have considered the analytical model, their applicable limits are restricted to more than 1 mm length in practical situations. Our new model considers both the sample and the sensor substrate thermal properties and is applicable to a sensor length less than 1 µm. In order to minimize the influence of the heat stimulus to the sample, the model formulates the required heat dissipating time for different sensor geometries. We propose fast and precise detection circuit architecture to realize our model, and we discuss the sensor performance for a number of different designs.

The mechanical properties are indispensable parameter for evaluate the 3 dimensional tissues. In this report, we developed an integrated micro vacuum chuck (IMVC) which could fix the three-dimensional multilayered tissue to tensile test system without stress concentrations and tensile test system for IMVC. Newly developed system can adjust vacuum pressure because IMVC was connected to vacuum pump and pressure sensor directory. The three-dimensional multilayered tissues were fabricated with technique of layer-by-layer (LbL) nanofilm coatings. By using the IMVC, we succeeded to measure tensile force of the three-dimensional multilayered tissues without stress concentrations. In the result, the maximum tensile force was about 410 μN (strain: 0.6). When strain reached 0.5, we measured tensile force about 350 μN and this value was same order of conventional study. Therefore, it was considered that the IMVC was useful tool for fixing 3D tissues in measurement of mechanical properties of 3D tissue.