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2011 International Symposium on Micro-NanoMechatronics and Human Science最新文献

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Dynamic transformation of a cell-sized liposome containing ganglioside 含有神经节苷脂的细胞大小脂质体的动态转化
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102232
Shruti Dhingra, Masamune Morita, Tsuyoshi Yoda, Mun'delanji C. Vestergaard, T. Hamada, M. Takagi
It is important to understand the physicochemical mechanisms that are responsible for the morphological changes in the cell membrane in the presence of various stimuli such as osmotic pressure. Role of micro domains such as lipid rafts in cellular processes is now beginning to unfold. In this study, we examined how constituent molecules affect the dynamical movement of liposomes. We used cell sized lipid vesicles to enable direct observation of these changes. We observed the effect of ganglioside (GM1) to osmotic stress induced membrane transformation in homogeneous and heterogeneous liposomes. Interestingly, it was observed that for the formation of sphero-stomatocyte there exists particular critical cut-off concentration. Also in the case of heterogeneous liposomes it was observed that at 10% molar ratio of GM1 almost all domains pinched out from the vesicles, forming their own homogeneous liposomes. Incorporation of GM1 into membrane leads to an increase of the line tension. Thus, necessary proteins can find themselves in one common raft and start the corresponding cascade of reactions.
了解在各种刺激(如渗透压)下细胞膜形态变化的物理化学机制是很重要的。微结构域如脂筏在细胞过程中的作用现在开始展开。在这项研究中,我们研究了组成分子如何影响脂质体的动态运动。我们使用细胞大小的脂质囊泡来直接观察这些变化。我们观察了神经节苷脂(GM1)对渗透胁迫诱导的均质和非均质脂质体膜转化的影响。有趣的是,我们观察到球形口细胞的形成存在特定的临界截止浓度。同样,在非均质脂质体的情况下,观察到在10%的GM1摩尔比下,几乎所有的结构域都从囊泡中挤出,形成它们自己的均质脂质体。GM1掺入膜中导致线张力增加。因此,必要的蛋白质可以发现自己在一个共同的筏,并开始相应的级联反应。
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引用次数: 2
Single molecule analysis of transcription factor-DNA complexes using atomic force microscopy 利用原子力显微镜对转录因子- dna复合物进行单分子分析
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102240
Masahiro Nakano, J. Teramoto, T. Shimada, Kaneyoshi Yamamoto, A. Ishihama
Transcription of the Escherichia coli genome is under the control of approximately 300 species of transcription factor. However, the regulatory roles remain unidentified for about 100 species of the transcription factor. As an approach to identify their regulation modes, we analyzed the dynamics of DNA-transcription factor interactions using the atomic force microscopy (AFM). Here we examined the DNA-binding mode of two types of transcription factor in the presence and absence of RNA polymerase: a novel growth-specific nucleoid protein Dan (DNA-binding protein under anaerobic growth conditions); and global regulators, CRP (cAMP receptor protein) and Cra (catabolite repressor and activator), for control of transport and metabolism of carbon sources.
大肠杆菌基因组的转录受大约300种转录因子的控制。然而,大约100种转录因子的调控作用仍未确定。为了确定它们的调控模式,我们使用原子力显微镜(AFM)分析了dna -转录因子相互作用的动力学。在这里,我们研究了两种类型的转录因子在存在和不存在RNA聚合酶的情况下的dna结合模式:一种新的生长特异性类核蛋白Dan(厌氧生长条件下的dna结合蛋白);以及控制碳源运输和代谢的全球调节因子,CRP (cAMP受体蛋白)和Cra(分解代谢抑制因子和激活因子)。
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引用次数: 0
Spheroid array formation by non-label cell manipulation using magneto-Archimedes effect 利用磁阿基米德效应的非标记细胞操作形成球体阵列
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102156
Y. Akiyama, K. Morishima
This paper demonstrates spheroid array formation by a label-free three-dimensional (3D) cell assembly method which is based on the magneto-Archimedes effect. A paramagnetic salt (Gd-DOTA) is added to the culturing medium to enhance the diamagnetic property of the cells, by which label-free magnetic manipulation becomes possible. In this study, a chamber including cells suspended in the paramagnetic medium was placed on a magnet array, and spots with low (that is, almost zero) magnetic flux densities were formed on the array with the magnet array, on which the cells aggregated. These cell aggregates became spheroids after one day of culture. The result suggests that this 3D cell assembly method is a promising approach to construct micro 3D tissues in a closed small space.
本文演示了一种基于磁阿基米德效应的无标记三维(3D)单元组装方法形成球体阵列。在培养基中加入顺磁盐(Gd-DOTA)以增强细胞的抗磁性,从而使无标记磁操作成为可能。在本研究中,将悬浮在顺磁介质中的细胞放置在磁体阵列上,在磁体阵列上形成低(即几乎为零)磁通密度的斑点,细胞聚集在该磁体阵列上。这些细胞聚集体在培养一天后变成球形。结果表明,这种三维细胞组装方法是在封闭的小空间中构建微型三维组织的一种很有前途的方法。
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引用次数: 4
Giant liposomes as microcapsules with large trapping volumes: Downsizing through various membrane filters and analysis with a calcein quenching method 巨脂质体作为具有大捕获体积的微胶囊:通过各种膜过滤器缩小尺寸并使用钙黄蛋白淬火方法进行分析
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102228
K. Tsumoto, Yuki Nakamura, M. Yamazoe, M. Tomita
Giant unilamellar vesicles (GUVs), or giant liposomes, are very similar to actual cells about the size (∼1 – 100 µm); therefore, GUVs have much larger trapping volumes than conventional liposomes (large unilamellar vesicles, LUVs; ∼100 nm). Albeit having such large trapping volumes, GUVs are rarely employed as microcapsules for biomedical purposes because of their difficulties of the preparation in physiological buffer solutions in large quantities with the narrow size distributions. Here, we comparatively downsized giant liposomes by making them filtrated through various membrane filters including polycarbonate, polytetrafluoroethylene and nitrocellulose/cellurose acetate membranes, and measured their trapping volumes using a calcein quenching method. Any filters with around 1–5 µm pore sizes were useful to reduce the size from more than 10 µm to less than 5 µm, and their trapping volumes were estimated to be around ten times higher than those of 100-nm LUVs, which are reasonable values; however, somewhat large fractions of vesicles that were much smaller than the pore sizes remained in the sample.
巨型单层囊泡(GUVs)或巨型脂质体的大小与实际细胞非常相似(约1 - 100µm);因此,guv具有比传统脂质体(大单层囊泡,luv;∼100海里)。尽管具有如此大的捕获体积,但由于guv难以在大量生理缓冲溶液中制备且尺寸分布狭窄,因此很少用作生物医学用途的微胶囊。本研究通过聚碳酸酯、聚四氟乙烯和硝化纤维素/醋酸纤维素膜等不同的膜过滤器对巨脂质体进行过滤,并采用钙黄蛋白猝灭法测量巨脂质体的捕获体积。任何孔径在1-5微米左右的过滤器都可以将孔径从大于10微米减小到小于5微米,其捕获体积估计比100纳米luv高10倍左右,这是合理的值;然而,比孔隙大小小得多的囊泡仍然存在于样品中。
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引用次数: 0
Inchworm type microrobot using friction force control mechanisms 采用摩擦力控制机构的尺蠖型微型机器人
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102192
Yukihiro Itatsu, A. Torii, A. Ueda
We studied inchworm type microrobots using piezoelectric elements. When the microrobot moves, friction between the microrobot and an operation surface disturbs the displacement of the microrobot. We propose the inchworm type microrobot which can control the friction force. The friction force is controlled by the use of squeeze film effect. The microrobot is composed of three friction force control mechanisms, two horizontal piezoelectric elements, and leaf springs. The friction force control mechanism is composed of a metal plate, a weight, and a stacked type piezoelectric element. When a vertical piezoelectric element vibrates vertically, the squeeze film effect occurs and the friction force control mechanism levitates. The leaf springs are used as vibration isolators. The friction force control mechanism levitating in air moves by the push and pull of the horizontal piezoelectric elements. In this study, first, the height of a bottom plate of the levitation mechanism is measured. Then we changed the control frequency and waveforms, and measured the velocity of the microrobot.
我们研究了利用压电元件的尺蠖型微型机器人。当微机器人运动时,微机器人与操作表面之间的摩擦会干扰微机器人的位移。提出了一种能够控制摩擦力的尺蠖型微型机器人。摩擦力是利用挤膜效应来控制的。该微型机器人由三个摩擦力控制机构、两个水平压电元件和钢板弹簧组成。摩擦力控制机构由金属板、砝码和堆叠式压电元件组成。当垂直压电元件垂直振动时,产生挤压膜效应,摩擦力控制机构悬浮。钢板弹簧用作隔振器。悬浮在空气中的摩擦力控制机构依靠水平压电元件的推拉运动。在本研究中,首先测量了悬浮机构底板的高度。然后改变控制频率和波形,测量微型机器人的速度。
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引用次数: 2
Micro droplet generation using micropore plates oscillated by ultrasonic torsional transducers 利用超声扭转换能器振动微孔板产生微液滴
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102165
Yusuke Kiyama, Yoshiyuki Tominaga, T. Kanda, K. Suzumori, Yoshiaki Yamada, Norihisa Seno
Monodisperse droplets have been an important subject in fields such as electronic materials, cosmetics productions and medical science. The goal of this study is to generate micro and uniform droplets in various viscosity liquids by using a micropore plate oscillated by an ultrasonic torsional transducer. When the micropore plate was not oscillated, droplets were generated irregularly. On the other hand, when the micropore plate was oscillated by the ultrasonic torsional transducer, droplets were generated regularly by ruffling liquid surface. We have generated droplets in the air by using pure water and silicone oil of various viscosities and evaluated droplets. We have designed the ultrasonic torsional transducer and a droplet generation device to apply a high pressure for the use of higher viscosity liquids. We have succeeded in generating droplets by using pure water when we used the ultrasonic torsional transducer. The average diameter of droplets was 30.0µm and the standard deviation was 0.98. We have also succeeded in generating 50cSt silicone oil droplets using the droplet generation device. The average diameter of droplets was 98.0µm and the standard deviation was 0.76.
单分散液滴已成为电子材料、化妆品生产和医学等领域的重要研究课题。本研究的目的是利用超声扭转换能器振荡的微孔板,在不同粘度的液体中产生微小而均匀的液滴。当微孔板不振荡时,液滴产生不规则。另一方面,当超声扭转换能器振荡微孔板时,液体表面皱折产生有规律的液滴。我们用纯净水和不同粘度的硅油在空气中产生液滴,并对液滴进行了评价。我们设计了超声波扭转换能器和液滴产生装置,用于高粘度液体的高压应用。我们在使用超声波扭转换能器时,成功地利用纯水产生了液滴。液滴平均直径为30.0µm,标准差为0.98。我们还利用液滴生成装置成功生成了50cSt的硅油液滴。液滴平均直径为98.0µm,标准差为0.76。
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引用次数: 3
Development of observation system to investigate both intracellular calcium concentration and mechanical stimuli to mammalian embryos 研究哺乳动物胚胎细胞内钙浓度和机械刺激的观察系统的建立
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102167
K. Matsuura, Koyo Watanabe, Mieko Kodama, Yuka Kuroda, K. Naruse
Using an air-actuating device, we investigated the cellular response to mechanical stimuli (MS) in mouse blastocysts. Both MS and intracellular calcium concentration ([Ca2+]i) were quantified based on time-resolved confocal microscopy images in the polydimethylsiloxane (PDMS) microfluidic channels by deforming a 0.1-mm membrane. [Ca2+]i was measured in a stained mouse embryo with Fluo-4 AM using confocal fluorescence microscopy. We captured a z-series stack of sections encompassing the entire embryo. When translocation velocities of the embryo and shear stress were 40 µm/s and 0.01 dyne/cm2, respectively, a 10% increase in the sum of fluorescent intensities (FI) was observed. When blastocysts were compressed, FI also increased in response to the applied MS. Compressive force estimated from the shape of the blastocysts was approximately 0.5–2.0 µN according to a force deformation curve for the mouse embryo. The average FI and sum of FIs increased by a factor of 1.1–1.2 times compared with those observed before MS. The increase in the sum of FI indicated that enhancement of [Ca2+]i would be induced by these MS.
利用空气驱动装置,研究了小鼠囊胚对机械刺激的细胞反应。通过形变0.1 mm的膜,利用时间分辨共聚焦显微镜图像定量测定聚二甲基硅氧烷(PDMS)微流控通道内的质谱和细胞内钙浓度([Ca2+]i)。用共聚焦荧光显微镜用Fluo-4 AM在染色的小鼠胚胎中测量[Ca2+]i。我们捕获了包含整个胚胎的z系列切片。当胚胎的移位速度和剪应力分别为40µm/s和0.01 dyne/cm2时,荧光强度(FI)总和增加10%。当囊胚被压缩时,FI也随着施加的ms而增加。根据小鼠胚胎的力变形曲线,囊胚形状估计的压缩力约为0.5-2.0µN。与MS前相比,平均FI和FI之和增加了1.1-1.2倍,FI之和的增加表明MS可诱导[Ca2+]i的增强。
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引用次数: 2
Novel device for transplantation of cell sheet and evaluation of thin polymer films by atomic force microscopy 一种新的细胞片移植装置及原子力显微镜对聚合物薄膜的评价
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102201
R. Takeuchi, K. Fukumori, Katsuhisa Sakaguchi, Y. Terajima, Tatsuya Shimizu, T. Okano, M. Umezu
Cell therapy is expected to a new tool to treat refractory diseases. In heart regeneration, it has been firstly conducted with needle injection of cell suspensions. Recently, cell sheet engineering emerged as another method of cell therapy. Cell sheet is prepared with a temperature responsive dish by temperature reduction. It is a thin-patch-like tissue construct and its thickness is several tens of micrometers. It is composed of cells and intrinsic extra cellular matrix only. The transplantation of the cell sheet has been already conducted in animal experiments and even in clinical trials. The cell sheet is transplanted at the surface of the heart, but it is difficult to transplant the cell sheet under the beating heart. To overcome this difficulty, we designed a device that was composed of two thin polymer films that have different friction. The films were made of polyurethane, polyethylene, or polypropylene. The cell sheet was set up on the device by sandwiching it with the less frictional film and the more frictional film. In this paper, using two different films having the different friction, the cell sheet was successfully transplanted to the static round polymer surface, the harvested heart, and even the beating heart of pig by removing the films step by step using the difference in friction. Also, surface properties such as friction, adhesion force and roughness of the films were studied by an atomic force microscopy (AFM). From the results of the study, the friction of the film was found to be likely proportional to the adhesion force and the inverse of roughness.
细胞疗法有望成为治疗难治性疾病的新工具。在心脏再生方面,首次采用针注射细胞悬浮液的方法进行。最近,细胞片工程作为细胞治疗的另一种方法出现了。细胞片是用温度反应盘通过温度还原法制备的。它是一种薄片状组织结构,其厚度为几十微米。它仅由细胞和固有的细胞外基质组成。细胞片的移植已经在动物实验甚至临床试验中进行。细胞片移植在心脏表面,但很难移植到跳动的心脏下。为了克服这一困难,我们设计了一种由两种具有不同摩擦的聚合物薄膜组成的装置。这些薄膜由聚氨酯、聚乙烯或聚丙烯制成。将电池片与摩擦较小的膜和摩擦较大的膜夹在一起,建立在设备上。本文采用两种摩擦力不同的膜,利用摩擦力的差异,逐步去除膜,将细胞片移植到静止的圆形聚合物表面、收获的心脏,甚至是跳动的猪心脏上。同时,利用原子力显微镜(AFM)研究了薄膜的摩擦、附着力和粗糙度等表面特性。从研究结果来看,薄膜的摩擦力可能与附着力成正比,与粗糙度成反比。
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引用次数: 0
Virtual building blocks using a 2.5D-display generating of tactile and force sensations 使用2.5 d显示器产生触觉和力感的虚拟积木
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102219
S. Tsuboi, M. Ohka
To develop new equipment for virtual reality, we study a combined display capable of stimulating the muscles and tendons of the forearms and tactile receptors in fingers to investigate tactile and force effects on simultaneous presentation. We arranged the handle of the display like a mouse so that anyone could comfortably use it without a priori knowledge. The present display is comprised of a master arm with a tactile display with a 4-by-12 array of stimulus pins driven by micro-actuators and an articulated manipulator to stimulate the muscles and tendons of the forearms and tactile receptors in fingers. Vertical movement of the virtual tactile pad in the virtual world is controlled in proportion to compressive force applied on the display pad, so that operators don't get tired during manipulations in 3D-space because their hands are supported by the manipulator. The compressive force is measured by a pressure sensor installed under the actuator array. Operators can work in 3D-space in spite of their manipulations in 2.5D-space. Evaluation experiments are performed using the display to verify its presentation capability. In the experiments, subjects try to build up virtual blocks in a line using the display. A precise building task is impossible for the force-only display because of no relative motion between the hands and the blocks, while the building task is completed by the combined display of tactile and force sensations with higher precision.
为了开发新的虚拟现实设备,我们研究了一种能够刺激前臂肌肉和肌腱以及手指触觉感受器的组合显示器,以研究触觉和力对同时呈现的影响。我们把显示器的手柄设计得像鼠标一样,这样任何人都可以在没有先验知识的情况下轻松地使用它。目前的显示器由一个带有触觉显示器的主臂和一个由微致动器驱动的4 × 12的刺激针阵列,以及一个关节操纵器组成,用于刺激前臂的肌肉和肌腱以及手指的触觉感受器。虚拟触觉垫在虚拟世界中的垂直运动与施加在显示垫上的压缩力成比例控制,因此在3d空间中操作时,由于操作者的手有机械手的支撑,因此不会感到疲劳。压缩力由安装在执行器阵列下的压力传感器测量。操作员可以在3d空间工作,尽管他们在2.5 d空间操作。利用该显示器进行了评估实验,以验证其显示能力。在实验中,受试者尝试用显示器在一条线上构建虚拟块。由于手和积木之间没有相对运动,单纯的力显示无法精确的完成建造任务,而触觉和力感觉的结合显示则具有更高的精度。
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引用次数: 0
Fabrication of a dynamic compression stimulus microdevice to cells for evaluating real-time cellular response 一种用于评估实时细胞反应的细胞动态压缩刺激微装置的制造
Pub Date : 2011-12-15 DOI: 10.1109/MHS.2011.6102212
Y. Nakashima, Y. Yang, K. Minami
This paper presents a microdevice intended to evaluate the real-time response of cells to compression stimulus for dynamic cell differentiation induction. This device was realized by the micro three-dimensional structure fabrication process using multiple exposures to photoresist. The microdevice consists of microchannels, a pressure inlet port, cell culture chambers and a diaphragm on the culture chamber for applying compressive pressure to cells. Compression stimulus to cells can be controlled by regulating of the diaphragm expansion with the pressure control. This device can make it to observe the behavior of cellular response stimulated by compressive pressure because this device is made of transparency materials and stimulates the cells without deforming the cell culture surface observed by an optical microscope. We demonstrated the validity of the fabrication process, and evaluated the performance of the test device. As a result of operation test, the diaphragm was deformed quickly by applied negative/positive pressure, and the diaphragm displacement became larger with increasing of the applied pressure. These results indicate that this device can control the intensity and the profile of cell stimulus by regulating the applied pressure.
本文提出了一种微型装置,用于评估细胞对压缩刺激的实时反应,以动态诱导细胞分化。该器件采用多次曝光光刻胶的微三维结构制造工艺实现。该微装置包括微通道、压力入口、细胞培养室和用于向细胞施加压缩压力的培养室上的隔膜。对细胞的压缩刺激可以通过控制压力来调节隔膜的膨胀来控制。由于该装置由透明材料制成,在不使光学显微镜观察到的细胞培养表面变形的情况下对细胞进行刺激,因此可以观察到细胞在压缩压力刺激下的反应行为。我们证明了制造工艺的有效性,并评估了测试装置的性能。运行试验结果表明,负压/正压作用下膜片变形快,且膜片位移随压力的增大而增大。这些结果表明,该装置可以通过调节施加的压力来控制细胞刺激的强度和轮廓。
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引用次数: 0
期刊
2011 International Symposium on Micro-NanoMechatronics and Human Science
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