247 results on '"Sakuma, Shinya"'
Search Results
202. Active size controlled on-chip droplet dispensing by magnetically driven microtool
- Author
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Yamanishi, Yoko, primary, Kihara, Yuki, additional, Sakuma, Shinya, additional, and Arai, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
203. T1501-2-4 Continuous Particle Separation Using Magnetically Driven Microtool in Microfluidic Chip
- Author
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MARUYAMA, Hisataka, primary, SAKUMA, Shinya, additional, CHAPURLAT, Benoit, additional, YAMANISHI, Yoko, additional, and ARAI, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
204. T1501-2-3 On-chip Active Droplet Dispensing by Hybrid Magnetically Driven Microtool
- Author
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YAMANISHI, Yoko, primary, KIHARA, Yuki, additional, SAKUMA, Shinya, additional, and ARAI, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
205. 2A2-L02 Magnetically Driven Robot-on-a-chip (MDRoC) : Part 2: Design and Fabrication of Hybrid MMT
- Author
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KIHARA, Yuki, primary, YAMANISHI, Yoko, additional, SAKUMA, Shinya, additional, and ARAI, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
206. 2A2-K08 All-in-One Unified Microfluidic Chip for Automation of Embryonic Cell Manipulation Based on Micro-Robotics
- Author
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Sakuma, Shinya, primary, Yamanishi, Yoko, additional, Arai, Fumihito, additional, Arai, Tatsuo, additional, Hasegawa, Akiyuki, additional, Tanikawa, Tamio, additional, Ichikawa, Akihiko, additional, Satoh, Osamu, additional, Nakayama, Akihiro, additional, Aso, Hiroshi, additional, Goto, Mitsuhiro, additional, Takahashi, Seiya, additional, and Matsukawa, Kazutsugu, additional
- Published
- 2009
- Full Text
- View/download PDF
207. Sorting of Micro-particles using Magnetically Driven Micro-tools
- Author
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Yamanishi, Yoko, primary, Sakuma, Shinya, additional, Onda, Kazuhisa, additional, and Arai, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
208. 2A2-L01 Magnetically Driven Robot-on-a-chip (MDRoC) : Part 3: Continuous Microparticle Filtration Using Magnetically Driven Microtool
- Author
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MARUYAMA, Hisataka, primary, SAKUMA, Shinya, additional, CHAPURLAT, Benoit, additional, YAMANISHI, Yoko, additional, and ARAI, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
209. 2A2-L03 Magnetically Driven Robot-on-a-chip (MDRoC) : Part 1: On-chip Particle Manipulations using 2DOF MMT
- Author
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YAMANISHI, Yoko, primary, SAKUMA, Shinya, additional, KIHARA, Yuki, additional, and ARAI, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
210. 2A1-J12 On-chip Micro-droplet Dispenser with Disposable Structure
- Author
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MIZUNUMA, Takehito, primary, SAKUMA, Shinya, additional, YAMANISHI, Yoko, additional, MARUYAMA, Hisataka, additional, and ARAI, Fumihito, additional
- Published
- 2009
- Full Text
- View/download PDF
211. Polymer-Based Magnetically Driven Microtool for On-Chip Particle Sorting
- Author
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Sakuma, Shinya, primary, Yamanishi, Yoko, additional, and Arai, Fumihito, additional
- Published
- 2008
- Full Text
- View/download PDF
212. On-chip cell manipulation by magnetically modified soft microactuators
- Author
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Yamanishi, Yoko, primary, Sakuma, Shinya, additional, and Arai, Fumihito, additional
- Published
- 2008
- Full Text
- View/download PDF
213. 2P1-D14 On-chip cell manipulation systems : Part 7: Development of magnetically driven microtool composed of permanent magnet and microparticle manipulation
- Author
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SAKUMA, Shinya, primary, YAMANISHI, Yoko, additional, KIHARA, Yuki, additional, and ARAI, Fumihito, additional
- Published
- 2008
- Full Text
- View/download PDF
214. 2P1-D15 On-chip cell manipulation systems : Part5: Microparticle loading system by magnetically driven 3D microtool
- Author
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YAMANISHI, Yoko, primary, SAKUMA, Shinya, additional, and ARAI, Fumihito, additional
- Published
- 2008
- Full Text
- View/download PDF
215. 2P1-D13 On-chip cell manipulation systems : Part6: On-chip Droplet Dispensing by Magnetically Driven Microtool
- Author
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YAMANISHI, Yoko, primary, KIHARA, Yuki, additional, SAKUMA, Shinya, additional, and ARAI, Fumihito, additional
- Published
- 2008
- Full Text
- View/download PDF
216. Magnetically Modified Soft Micro Actuators for Oocyte Manipulation
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Yamanishi, Yoko, primary, Sakuma, Shinya, additional, and Arai, Fumihito, additional
- Published
- 2007
- Full Text
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217. Ising vectors and automorphism groups of commutant subalgebras related to root systems
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Lam, Ching Hung, primary, Sakuma, Shinya, additional, and Yamauchi, Hiroshi, additional
- Published
- 2006
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218. The McKay-Thompson Series of Some Elements of Order 3,5,and 7 in the Monster
- Author
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Sakuma, Shinya, primary
- Published
- 2003
- Full Text
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219. Vertex operator algebra with two Miyamoto involutions generating <f>S3</f>
- Author
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Sakuma, Shinya and Yamauchi, Hiroshi
- Subjects
- *
OPERATOR algebras , *AUTOMORPHISMS - Abstract
In this article we study a VOA with two Miyamoto involutions generating
S3 . In [math.GR/0112031], Miyamoto showed that a VOA generated by two conformal vectors whose Miyamoto involutions generate an automorphism group isomorphic toS3 is isomorphic to one of the four candidates he listed. We construct one of them and prove that our VOA is actually the same asVA(e,f) studied by Miyamoto. We also show that there is an embedding into the moonshine VOA. Using our VOA, we can define the 3A-triality of the Monster. [Copyright &y& Elsevier]- Published
- 2003
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220. The McKay-Thompson Series of Some Elements of Order 3, 5, and 7 in the Monster.
- Author
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Sakuma, Shinya
- Subjects
- *
AUTOMORPHISMS , *BIVECTORS , *FINITE simple groups , *GROUP theory - Abstract
In this article, we explicitly calculate the McKay-Thompson series corresponding to 3A, 7A, 3B, 5B and 15C elements of the Monster simple group by using the framed VOA structure of the Moonshine VOA and automorphisms which permute 48 mutually orthogonal rational conformal vectors with central charge 1/2. [ABSTRACT FROM AUTHOR]
- Published
- 2003
- Full Text
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221. Local Ablation by a Microelectric Knife: Enucleation of an Oocyte
- Author
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Yamanishi, Yoko, Kuriki, Hiroki, Sakuma, Shinya, Onda, Kazuhisa, and Arai, Fumihito
- Abstract
The electric knife is one of the indispensable surgical devices widely used in surgical operation. However, it has not been remarkably improved since its invention several decades ago. For example, thermal damage of the current radiofrequency electric knife, which is one of the most nonintrusive type of electric knives, is more than several hundred micrometers. A laser device, which was invented approximately 70 years ago, has been developed remarkably, and various research report the interaction between the cells and laser to fulfill the cellular-order ablation [1], [2].
- Published
- 2012
- Full Text
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222. 6-Transposition Property of τ-Involutions of Vertex Operator Algebras.
- Author
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Sakuma, Shinya
- Subjects
- *
VERTEX operator algebras , *OPERATOR algebras , *NONASSOCIATIVE algebras , *VECTOR algebra , *ISOMORPHISM (Mathematics) , *COMMUTATIVE algebra - Abstract
In this paper, we study the subalgebra generated by two Ising vectors in the Griess algebra of a vertex operator algebra. We show that its structure is uniquely determined by some inner products of Ising vectors. We prove that the order of the product of two τ-involutions is less than or equal to 6, and we determine the inner product of two Ising vectors. [ABSTRACT FROM PUBLISHER]
- Published
- 2007
- Full Text
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223. Spatiotemporally controlled microvortices provide advanced microfluidic components.
- Author
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Saito M, Arai F, Yamanishi Y, and Sakuma S
- Abstract
Microvortices are emerging components that impart functionality to microchannels by exploiting inertia effects such as high shear stress, effective fluid diffusion, and large pressure loss. Exploring the dynamic generation of vortices further expands the scope of microfluidic applications, including cell stimulation, fluid mixing, and transport. Despite the crucial role of vortices' development within sub-millisecond timescales, previous studies in microfluidics did not explore the modulation of the Reynolds number (Re) in the range of several hundred. In this study, we modulated high-speed flows (54 < [Formula: see text] < 456) within sub-millisecond timescales using a piezo-driven on-chip membrane pump. By applying this method to microchannels with asymmetric geometries, we successfully controlled the spatiotemporal development of vortices, adjusting their behavior in response to oscillatory flow directions. These different vortices induced different pressure losses, imparting the microchannels with direction-dependent flow resistance, mimicking a diode-like behavior. Through precise control of vortex development, we managed to regulate this direction-dependent resistance, enabling the rectification of oscillatory flow resembling a diode and the ability to switch its rectification direction. This component facilitated bidirectional flow control without the need for mechanical valves. Moreover, we demonstrated its application in microfluidic cell pipetting, enabling the isolation of single cells. Consequently, based on modulating high-speed flow, our approach offers precise control over the spatiotemporal development of vortices in microstructures, thereby introducing innovative microfluidic functionalities., Competing Interests: Competing interests statement:The authors declare no competing interest.
- Published
- 2024
- Full Text
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224. Optimization of QuEChERS Extraction for Determination of Carotenoids, Polyphenols, and Sterols in Orange Juice Using Design of Experiments and Response Surface Methodology.
- Author
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Iwasaki Y, Yamada S, Sakuma S, Kanba S, Youda C, Ono M, Ito R, Kamei J, and Akiyama H
- Abstract
Several compounds with different physical properties are present in foods, biological components, and environmental samples, and there are cases in which these must be analyzed simultaneously. However, it is difficult to extract compounds with different physical properties from the same sample using a single method. In the present study, we examined the optimal conditions for the QuEChERS extraction of several kinds of compounds from orange juice using design of experiments (DoE) and response surface methodology (RSM) to determine the optimal ratio of organic solvent to sodium chloride. We determined the optimal extraction conditions, which were within the design space, using 100% tetrahydrofuran (THF) as the extraction organic solvent and NaCl:MgSO
4 = 75:25 as the salt. The developed LC/MS/MS method using QuEChERS extraction achieved specific detection and precise quantification. Finally, we measured the polyphenols, sterols, and carotenoids in citrus juice using the optimized QuEChERS extraction method before LC/MS/MS analysis. Most of the analytes were quantifiable in orange juice. The optimized method achieved ease of operation, the extraction of analytes from food samples in a short time (within 30 min), minimization of analytical residues, and reliability. The DoE and RSM approach may contribute to better optimization of the extraction conditions for the lowest number of experiments.- Published
- 2023
- Full Text
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225. Viscosity-aided electromechanical poration of cells for transfecting molecules.
- Author
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Huang W, Sakuma S, Tottori N, Sugano SS, and Yamanishi Y
- Subjects
- Viscosity, Transfection, Microbubbles, Electricity
- Abstract
Cell poration technologies offer opportunities not only to understand the activities of biological molecules but also to investigate genetic manipulation possibilities. Unfortunately, transferring large molecules that can carry huge genomic information is challenging. Here, we demonstrate electromechanical poration using a core-shell-structured microbubble generator, consisting of a fine microelectrode covered with a dielectric material. By introducing a microcavity at its tip, we could concentrate the electrical field with the application of electric pulses and generate microbubbles for electromechanical stimulation of cells. Specifically, the technology enables transfection with molecules that are thousands of kDa even into osteoblasts and Chlamydomonas , which are generally considered to be difficult to inject. Notably, we found that the transfection efficiency can be enhanced by adjusting the viscosity of the cell suspension, which was presumably achieved by remodeling of the membrane cytoskeleton. The applicability of the approach to a variety of cell types opens up numerous emerging gene engineering applications.
- Published
- 2022
- Full Text
- View/download PDF
226. On-Demand Metallization System Using Micro-Plasma Bubbles.
- Author
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Yamashita Y, Sakuma S, and Yamanishi Y
- Abstract
3D wiring technology is required for the integration of micro-nano devices on various 3D surfaces. However, current wiring technologies cannot be adapted to a variety of materials and surfaces. Here, we propose a new metal deposition method using only a micro-plasma bubble injector and a metal ion solution. Micro-plasma bubbles were generated on demand using pulses, and the localized reaction field enables metal deposition independent of the substrate. Three different modes of micro-plasma bubble generation were created depending on the power supply conditions and mode suitable for metal deposition. Furthermore, using a mode in which one bubble was generated for all pulses among the three modes, copper deposition on dry/wet materials, such as chicken tissue and glass substrates, was achieved. In addition, metal deposition of copper, nickel, chromium, cobalt, and zinc was achieved by simply changing the metal ion solution. Finally, patterning on glass and epoxy resin was performed. Notably, the proposed metal deposition method is conductivity independent. The proposed method is a starting point for 3D wiring of wet materials, which is difficult with existing technologies. Our complete system makes it possible to directly attach sensors and actuators to living organisms and robots, for example, and contribute to soft robotics and biomimetics.
- Published
- 2022
- Full Text
- View/download PDF
227. Microfluidic Bioreactor Made of Cyclo-Olefin Polymer for Observing On-Chip Platelet Production.
- Author
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Kumon H, Sakuma S, Nakamura S, Maruyama H, Eto K, and Arai F
- Abstract
We previously proposed a microfluidic bioreactor with glass-Si-glass layers to evaluate the effect of the fluid force on platelet (PLT) production and fabricated a three-dimensional (3D) microchannel by combining grayscale photolithography and deep reactive ion etching. However, a challenge remains in observing the detailed process of PLT production owing to the low visibility of the microfluidic bioreactor. In this paper, we present a transparent microfluidic bioreactor made of cyclo-olefin polymer (COP) with which to observe the process of platelet-like particle (PLP) production under a bright-field, which allows us to obtain image data at a high sampling rate. We succeeded in fabricating the COP microfluidic bioreactor with a 3D microchannel. We investigated the bonding strength of COP-COP layers and confirmed the effectiveness of the microfluidic bioreactor. Results of on-chip PLP production using immortalized megakaryocyte cell lines (imMKCLs) derived from human-induced pluripotent stem cells show that the average total number of produced PLPs per imMKCL was 17.6 PLPs/imMKCL, which is comparable to that of our previous glass-Si-glass microfluidic bioreactor (17.4 PLPs/imMKCL). We succeeded in observing PLP production under a bright-field using the presented microfluidic bioreactor and confirmed that PLP fragmented in a narrow area of proplatelet-like protrusions.
- Published
- 2021
- Full Text
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228. Design of Electrohydrodynamic Devices with Consideration of Electrostatic Energy.
- Author
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Sato T, Sakuma S, Hijikuro M, Maeda S, Anyoji M, and Yamanishi Y
- Abstract
The importance of actuators that can be integrated with flexible robot structures and mechanisms has increased in recent years with the advance of soft robotics. In particular, electrohydrodynamic (EHD) actuators, which have expandable integrability to adapt to the flexible motion of soft robots, have received much attention in the field of soft robotics. Studies have deepened the understanding of steady states of EHD phenomena but nonsteady states are not well understood. We herein observe the development process of fluid in a microchannel adopting a Schlieren technique with the aid of a high-speed camera. In addition, we analyze the behavior of fluid flow in a microchannel that is designed to have pairs of parallel plate electrodes adopting a computational fluid dynamics technique. Results indicate the importance of considering flow generated by electrostatic energy, which tends to be ignored in constructing and evaluating EHD devices, and by the body force generated by the ion-drag force. By considering these effects, we estimate the development process of EHD flow and confirm the importance of considering the generation of vortices and their interactions inside the microchannel during the development of EHD devices., Competing Interests: The authors declare that there are no conflicts of interest regarding the publication of this article., (Copyright © 2021 Tasuku Sato et al.)
- Published
- 2021
- Full Text
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229. Raman image-activated cell sorting.
- Author
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Nitta N, Iino T, Isozaki A, Yamagishi M, Kitahama Y, Sakuma S, Suzuki Y, Tezuka H, Oikawa M, Arai F, Asai T, Deng D, Fukuzawa H, Hase M, Hasunuma T, Hayakawa T, Hiraki K, Hiramatsu K, Hoshino Y, Inaba M, Inoue Y, Ito T, Kajikawa M, Karakawa H, Kasai Y, Kato Y, Kobayashi H, Lei C, Matsusaka S, Mikami H, Nakagawa A, Numata K, Ota T, Sekiya T, Shiba K, Shirasaki Y, Suzuki N, Tanaka S, Ueno S, Watarai H, Yamano T, Yazawa M, Yonamine Y, Di Carlo D, Hosokawa Y, Uemura S, Sugimura T, Ozeki Y, and Goda K
- Subjects
- Animals, Humans, Cell Separation methods, Spectrum Analysis, Raman methods
- Abstract
The advent of image-activated cell sorting and imaging-based cell picking has advanced our knowledge and exploitation of biological systems in the last decade. Unfortunately, they generally rely on fluorescent labeling for cellular phenotyping, an indirect measure of the molecular landscape in the cell, which has critical limitations. Here we demonstrate Raman image-activated cell sorting by directly probing chemically specific intracellular molecular vibrations via ultrafast multicolor stimulated Raman scattering (SRS) microscopy for cellular phenotyping. Specifically, the technology enables real-time SRS-image-based sorting of single live cells with a throughput of up to ~100 events per second without the need for fluorescent labeling. To show the broad utility of the technology, we show its applicability to diverse cell types and sizes. The technology is highly versatile and holds promise for numerous applications that are previously difficult or undesirable with fluorescence-based technologies.
- Published
- 2020
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230. Intelligent image-activated cell sorting 2.0.
- Author
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Isozaki A, Mikami H, Tezuka H, Matsumura H, Huang K, Akamine M, Hiramatsu K, Iino T, Ito T, Karakawa H, Kasai Y, Li Y, Nakagawa Y, Ohnuki S, Ota T, Qian Y, Sakuma S, Sekiya T, Shirasaki Y, Suzuki N, Tayyabi E, Wakamiya T, Xu M, Yamagishi M, Yan H, Yu Q, Yan S, Yuan D, Zhang W, Zhao Y, Arai F, Campbell RE, Danelon C, Di Carlo D, Hiraki K, Hoshino Y, Hosokawa Y, Inaba M, Nakagawa A, Ohya Y, Oikawa M, Uemura S, Ozeki Y, Sugimura T, Nitta N, and Goda K
- Subjects
- Algorithms, Cell Separation, Flow Cytometry, Neural Networks, Computer, Software
- Abstract
The advent of intelligent image-activated cell sorting (iIACS) has enabled high-throughput intelligent image-based sorting of single live cells from heterogeneous populations. iIACS is an on-chip microfluidic technology that builds on a seamless integration of a high-throughput fluorescence microscope, cell focuser, cell sorter, and deep neural network on a hybrid software-hardware data management architecture, thereby providing the combined merits of optical microscopy, fluorescence-activated cell sorting (FACS), and deep learning. Here we report an iIACS machine that far surpasses the state-of-the-art iIACS machine in system performance in order to expand the range of applications and discoveries enabled by the technology. Specifically, it provides a high throughput of ∼2000 events per second and a high sensitivity of ∼50 molecules of equivalent soluble fluorophores (MESFs), both of which are 20 times superior to those achieved in previous reports. This is made possible by employing (i) an image-sensor-based optomechanical flow imaging method known as virtual-freezing fluorescence imaging and (ii) a real-time intelligent image processor on an 8-PC server equipped with 8 multi-core CPUs and GPUs for intelligent decision-making, in order to significantly boost the imaging performance and computational power of the iIACS machine. We characterize the iIACS machine with fluorescent particles and various cell types and show that the performance of the iIACS machine is close to its achievable design specification. Equipped with the improved capabilities, this new generation of the iIACS technology holds promise for diverse applications in immunology, microbiology, stem cell biology, cancer biology, pathology, and synthetic biology.
- Published
- 2020
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231. Author Correction: A practical guide to intelligent image-activated cell sorting.
- Author
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Isozaki A, Mikami H, Hiramatsu K, Sakuma S, Kasai Y, Iino T, Yamano T, Yasumoto A, Oguchi Y, Suzuki N, Shirasaki Y, Endo T, Ito T, Hiraki K, Yamada M, Matsusaka S, Hayakawa T, Fukuzawa H, Yatomi Y, Arai F, Di Carlo D, Nakagawa A, Hoshino Y, Hosokawa Y, Uemura S, Sugimura T, Ozeki Y, Nitta N, and Goda K
- Abstract
An amendment to this paper has been published and can be accessed via a link at the top of the paper.
- Published
- 2019
- Full Text
- View/download PDF
232. High-speed microparticle isolation unlimited by Poisson statistics.
- Author
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Iino T, Okano K, Lee SW, Yamakawa T, Hagihara H, Hong ZY, Maeno T, Kasai Y, Sakuma S, Hayakawa T, Arai F, Ozeki Y, Goda K, and Hosokawa Y
- Abstract
High-speed isolation of microparticles (e.g., microplastics, heavy metal particles, microbes, cells) from heterogeneous populations is the key element of high-throughput sorting instruments for chemical, biological, industrial and medical applications. Unfortunately, the performance of continuous microparticle isolation or so-called sorting is fundamentally limited by the trade-off between throughput, purity, and yield. For example, at a given throughput, high-purity sorting needs to sacrifice yield, or vice versa. This is due to Poisson statistics of events (i.e., microparticles, microparticle clusters, microparticle debris) in which the interval between successive events is stochastic and can be very short. Here we demonstrate an on-chip microparticle sorter with an ultrashort switching window in both time (10 μs) and space (10 μm) at a high flow speed of 1 m s
-1 , thereby overcoming the Poisson trade-off. This is made possible by using femtosecond laser pulses that can produce highly localized transient cavitation bubbles in a microchannel to kick target microparticles from an acoustically focused, densely aligned, bumper-to-bumper stream of microparticles. Our method is important for rare-microparticle sorting applications where both high purity and high yield are required to avoid missing rare microparticles.- Published
- 2019
- Full Text
- View/download PDF
233. Label-free chemical imaging flow cytometry by high-speed multicolor stimulated Raman scattering.
- Author
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Suzuki Y, Kobayashi K, Wakisaka Y, Deng D, Tanaka S, Huang CJ, Lei C, Sun CW, Liu H, Fujiwaki Y, Lee S, Isozaki A, Kasai Y, Hayakawa T, Sakuma S, Arai F, Koizumi K, Tezuka H, Inaba M, Hiraki K, Ito T, Hase M, Matsusaka S, Shiba K, Suga K, Nishikawa M, Jona M, Yatomi Y, Yalikun Y, Tanaka Y, Sugimura T, Nitta N, Goda K, and Ozeki Y
- Subjects
- Cell Line, Tumor, Humans, Microalgae cytology, Microalgae metabolism, Staining and Labeling, Flow Cytometry methods, Imaging, Three-Dimensional, Spectrum Analysis, Raman methods
- Abstract
Combining the strength of flow cytometry with fluorescence imaging and digital image analysis, imaging flow cytometry is a powerful tool in diverse fields including cancer biology, immunology, drug discovery, microbiology, and metabolic engineering. It enables measurements and statistical analyses of chemical, structural, and morphological phenotypes of numerous living cells to provide systematic insights into biological processes. However, its utility is constrained by its requirement of fluorescent labeling for phenotyping. Here we present label-free chemical imaging flow cytometry to overcome the issue. It builds on a pulse pair-resolved wavelength-switchable Stokes laser for the fastest-to-date multicolor stimulated Raman scattering (SRS) microscopy of fast-flowing cells on a 3D acoustic focusing microfluidic chip, enabling an unprecedented throughput of up to ∼140 cells/s. To show its broad utility, we use the SRS imaging flow cytometry with the aid of deep learning to study the metabolic heterogeneity of microalgal cells and perform marker-free cancer detection in blood., Competing Interests: Conflict of interest statement: Y.O., Y.S., Y.W., and K.G. are inventors of a patent application covering SRS imaging with a wavelength-switched laser. All other authors declare no conflict of interest., (Copyright © 2019 the Author(s). Published by PNAS.)
- Published
- 2019
- Full Text
- View/download PDF
234. A practical guide to intelligent image-activated cell sorting.
- Author
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Isozaki A, Mikami H, Hiramatsu K, Sakuma S, Kasai Y, Iino T, Yamano T, Yasumoto A, Oguchi Y, Suzuki N, Shirasaki Y, Endo T, Ito T, Hiraki K, Yamada M, Matsusaka S, Hayakawa T, Fukuzawa H, Yatomi Y, Arai F, Di Carlo D, Nakagawa A, Hoshino Y, Hosokawa Y, Uemura S, Sugimura T, Ozeki Y, Nitta N, and Goda K
- Subjects
- Cells, Cultured, Humans, Lab-On-A-Chip Devices, Microalgae cytology, Signal Processing, Computer-Assisted, Software, Flow Cytometry methods, Image Processing, Computer-Assisted methods, Neural Networks, Computer, Single-Cell Analysis methods
- Abstract
Intelligent image-activated cell sorting (iIACS) is a machine-intelligence technology that performs real-time intelligent image-based sorting of single cells with high throughput. iIACS extends beyond the capabilities of fluorescence-activated cell sorting (FACS) from fluorescence intensity profiles of cells to multidimensional images, thereby enabling high-content sorting of cells or cell clusters with unique spatial chemical and morphological traits. Therefore, iIACS serves as an integral part of holistic single-cell analysis by enabling direct links between population-level analysis (flow cytometry), cell-level analysis (microscopy), and gene-level analysis (sequencing). Specifically, iIACS is based on a seamless integration of high-throughput cell microscopy (e.g., multicolor fluorescence imaging, bright-field imaging), cell focusing, cell sorting, and deep learning on a hybrid software-hardware data management infrastructure, enabling real-time automated operation for data acquisition, data processing, intelligent decision making, and actuation. Here, we provide a practical guide to iIACS that describes how to design, build, characterize, and use an iIACS machine. The guide includes the consideration of several important design parameters, such as throughput, sensitivity, dynamic range, image quality, sort purity, and sort yield; the development and integration of optical, microfluidic, electrical, computational, and mechanical components; and the characterization and practical usage of the integrated system. Assuming that all components are readily available, a team of several researchers experienced in optics, electronics, digital signal processing, microfluidics, mechatronics, and flow cytometry can complete this protocol in ~3 months.
- Published
- 2019
- Full Text
- View/download PDF
235. Local traction force in the proximal leading process triggers nuclear translocation during neuronal migration.
- Author
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Umeshima H, Nomura KI, Yoshikawa S, Hörning M, Tanaka M, Sakuma S, Arai F, Kaneko M, and Kengaku M
- Subjects
- Actomyosin physiology, Animals, Biomechanical Phenomena, Cells, Cultured, Mice, Inbred ICR, Microscopy, Atomic Force, Cell Movement, Cell Nucleus physiology, Neurons physiology
- Abstract
Somal translocation in long bipolar neurons is regulated by actomyosin contractile forces, yet the precise spatiotemporal sites of force generation are unknown. Here we investigate the force dynamics generated during somal translocation using traction force microscopy. Neurons with a short leading process generated a traction force in the growth cone and counteracting forces in the leading and trailing processes. In contrast, neurons with a long leading process generated a force dipole with opposing traction forces in the proximal leading process during nuclear translocation. Transient accumulation of actin filaments was observed at the dipole center of the two opposing forces, which was abolished by inhibition of myosin II activity. A swelling in the leading process emerged and generated a traction force that pulled the nucleus when nuclear translocation was physically hampered. The traction force in the leading process swelling was uncoupled from somal translocation in neurons expressing a dominant negative mutant of the KASH protein, which disrupts the interaction between cytoskeletal components and the nuclear envelope. Our results suggest that the leading process is the site of generation of actomyosin-dependent traction force in long bipolar neurons, and that the traction force is transmitted to the nucleus via KASH proteins., (Copyright © 2018 Elsevier B.V. and Japan Neuroscience Society. All rights reserved.)
- Published
- 2019
- Full Text
- View/download PDF
236. High-throughput label-free molecular fingerprinting flow cytometry.
- Author
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Hiramatsu K, Ideguchi T, Yonamine Y, Lee S, Luo Y, Hashimoto K, Ito T, Hase M, Park JW, Kasai Y, Sakuma S, Hayakawa T, Arai F, Hoshino Y, and Goda K
- Subjects
- Carbon Dioxide metabolism, Carbon Isotopes metabolism, Chlorophyceae metabolism, Flow Cytometry instrumentation, Fourier Analysis, High-Throughput Screening Assays instrumentation, High-Throughput Screening Assays methods, Lab-On-A-Chip Devices, Photosynthesis, Reproducibility of Results, Single-Cell Analysis instrumentation, Single-Cell Analysis methods, Spectrum Analysis, Raman instrumentation, Vibration, Xanthophylls metabolism, Flow Cytometry methods, Spectrum Analysis, Raman methods
- Abstract
Flow cytometry is an indispensable tool in biology for counting and analyzing single cells in large heterogeneous populations. However, it predominantly relies on fluorescent labeling to differentiate cells and, hence, comes with several fundamental drawbacks. Here, we present a high-throughput Raman flow cytometer on a microfluidic chip that chemically probes single live cells in a label-free manner. It is based on a rapid-scan Fourier-transform coherent anti-Stokes Raman scattering spectrometer as an optical interrogator, enabling us to obtain the broadband molecular vibrational spectrum of every single cell in the fingerprint region (400 to 1600 cm
-1 ) with a record-high throughput of ~2000 events/s. As a practical application of the method not feasible with conventional flow cytometry, we demonstrate high-throughput label-free single-cell analysis of the astaxanthin productivity and photosynthetic dynamics of Haematococcus lacustris .- Published
- 2019
- Full Text
- View/download PDF
237. Intelligent Image-Activated Cell Sorting.
- Author
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Nitta N, Sugimura T, Isozaki A, Mikami H, Hiraki K, Sakuma S, Iino T, Arai F, Endo T, Fujiwaki Y, Fukuzawa H, Hase M, Hayakawa T, Hiramatsu K, Hoshino Y, Inaba M, Ito T, Karakawa H, Kasai Y, Koizumi K, Lee S, Lei C, Li M, Maeno T, Matsusaka S, Murakami D, Nakagawa A, Oguchi Y, Oikawa M, Ota T, Shiba K, Shintaku H, Shirasaki Y, Suga K, Suzuki Y, Suzuki N, Tanaka Y, Tezuka H, Toyokawa C, Yalikun Y, Yamada M, Yamagishi M, Yamano T, Yasumoto A, Yatomi Y, Yazawa M, Di Carlo D, Hosokawa Y, Uemura S, Ozeki Y, and Goda K
- Subjects
- Animals, Deep Learning, Humans, Flow Cytometry methods, High-Throughput Screening Assays methods, Image Processing, Computer-Assisted methods
- Abstract
A fundamental challenge of biology is to understand the vast heterogeneity of cells, particularly how cellular composition, structure, and morphology are linked to cellular physiology. Unfortunately, conventional technologies are limited in uncovering these relations. We present a machine-intelligence technology based on a radically different architecture that realizes real-time image-based intelligent cell sorting at an unprecedented rate. This technology, which we refer to as intelligent image-activated cell sorting, integrates high-throughput cell microscopy, focusing, and sorting on a hybrid software-hardware data-management infrastructure, enabling real-time automated operation for data acquisition, data processing, decision-making, and actuation. We use it to demonstrate real-time sorting of microalgal and blood cells based on intracellular protein localization and cell-cell interaction from large heterogeneous populations for studying photosynthesis and atherothrombosis, respectively. The technology is highly versatile and expected to enable machine-based scientific discovery in biological, pharmaceutical, and medical sciences., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
238. Microfluidic single-particle chemical analyzer with dual-comb coherent Raman spectroscopy.
- Author
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Ideguchi T, Nakamura T, Takizawa S, Tamamitsu M, Lee S, Hiramatsu K, Ramaiah-Badarla V, Park JW, Kasai Y, Hayakawa T, Sakuma S, Arai F, and Goda K
- Abstract
Label-free particle analysis is a powerful tool in chemical, pharmaceutical, and cosmetic industries as well as in basic sciences, but its throughput is significantly lower than that of fluorescence-based counterparts. Here we present a label-free single-particle analyzer based on broadband dual-comb coherent Raman scattering spectroscopy operating at a spectroscopic scan rate of 10 kHz. As a proof-of-concept demonstration, we perform broadband coherent anti-Stokes Raman scattering measurements of polystyrene microparticles flowing on an acoustofluidic chip at a high throughput of >1000 particles per second. This high-throughput label-free particle analyzer has the potential for high-precision statistical analysis of a large number of microparticles including biological cells.
- Published
- 2018
- Full Text
- View/download PDF
239. Turbulence Activates Platelet Biogenesis to Enable Clinical Scale Ex Vivo Production.
- Author
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Ito Y, Nakamura S, Sugimoto N, Shigemori T, Kato Y, Ohno M, Sakuma S, Ito K, Kumon H, Hirose H, Okamoto H, Nogawa M, Iwasaki M, Kihara S, Fujio K, Matsumoto T, Higashi N, Hashimoto K, Sawaguchi A, Harimoto KI, Nakagawa M, Yamamoto T, Handa M, Watanabe N, Nishi E, Arai F, Nishimura S, and Eto K
- Subjects
- Bioreactors, Cell Culture Techniques instrumentation, Humans, Hydrodynamics, Induced Pluripotent Stem Cells metabolism, Megakaryocytes metabolism, Megakaryocytes physiology, Blood Platelets metabolism, Cell Culture Techniques methods, Thrombopoiesis physiology
- Abstract
The ex vivo generation of platelets from human-induced pluripotent cells (hiPSCs) is expected to compensate donor-dependent transfusion systems. However, manufacturing the clinically required number of platelets remains unachieved due to the low platelet release from hiPSC-derived megakaryocytes (hiPSC-MKs). Here, we report turbulence as a physical regulator in thrombopoiesis in vivo and its application to turbulence-controllable bioreactors. The identification of turbulent energy as a determinant parameter allowed scale-up to 8 L for the generation of 100 billion-order platelets from hiPSC-MKs, which satisfies clinical requirements. Turbulent flow promoted the release from megakaryocytes of IGFBP2, MIF, and Nardilysin to facilitate platelet shedding. hiPSC-platelets showed properties of bona fide human platelets, including circulation and hemostasis capacities upon transfusion in two animal models. This study provides a concept in which a coordinated physico-chemical mechanism promotes platelet biogenesis and an innovative strategy for ex vivo platelet manufacturing., (Copyright © 2018 Elsevier Inc. All rights reserved.)
- Published
- 2018
- Full Text
- View/download PDF
240. Measurement of the mechanical properties of single Synechocystis sp. strain PCC6803 cells in different osmotic concentrations using a robot-integrated microfluidic chip.
- Author
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Chang D, Sakuma S, Kera K, Uozumi N, and Arai F
- Subjects
- Biomechanical Phenomena physiology, Cell Line, Elastic Modulus, Equipment Design, Single-Cell Analysis methods, Lab-On-A-Chip Devices, Optical Tweezers, Osmotic Pressure physiology, Robotics instrumentation, Single-Cell Analysis instrumentation, Synechocystis cytology, Synechocystis physiology
- Abstract
Synechocystis sp. strain PCC6803 (Synechocystis) is a model microorganism and its mechanosensitive (MS) channels play important roles in its osmoadaptation mechanism. When the osmotic concentration of the culture environment changes, the inner pressure of the cell also changes due to the transportation of water through ion channels. Because the tension in the cell membrane relates to the inner pressure, we expect that the response of the MS channels to an osmotic concentration change could be evaluated by measuring their mechanical properties. Here, we propose a system for the measurement of the mechanical properties of a single Synechocystis cell. We developed a robot-integrated microfluidic chip combined with optical tweezers. The chip has an external actuated pushing probe and a force sensor probe. A single cell was located between the tip of both probes using the optical tweezers and was then deformed using the probes. As a result, we could measure the force and deformation and compare the Young's moduli of two groups: a group of wild type cells and a group of mutant (genetically modified) cells with a defect in the MS channels, at three different osmotic concentrations. The results showed that the Young's modulus of each group changed according to the osmotic concentration, while changes in cell size were too small to be detected. These results confirmed that the proposed evaluation method provides an understanding of the physiological function of MS channels for keeping the cell integrity of microorganisms when the cells are exposed to different external osmotic changes.
- Published
- 2018
- Full Text
- View/download PDF
241. On-chip cell sorting by high-speed local-flow control using dual membrane pumps.
- Author
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Sakuma S, Kasai Y, Hayakawa T, and Arai F
- Subjects
- Equipment Design, Euglena gracilis cytology, Flow Cytometry methods, Microfluidic Analytical Techniques methods, Microscopy, Fluorescence, Flow Cytometry instrumentation, Lab-On-A-Chip Devices, Microfluidic Analytical Techniques instrumentation
- Abstract
Although researchers have proposed various methods of on-chip cell sorting, high-throughput sorting of large cells remains hampered by the difficulty of controlling high-speed flow over a wide sorting area. To overcome this problem, we proposed high-speed local-flow control using dual membrane pumps driven by piezoelectric actuators placed on the outside of a microfluidic chip in this paper. We evaluated the controllability of shifting the flow profile by the local-flow. The results indicated that we could sort large cells up to approximately 150 μm in size with an equivalent throughput of 31 kHz. Because our method can control the flow profiles, it is applicable not only to large cells but also to small cells. The cell-sorting efficacy of the proposed method was experimentally evaluated on Euglena gracilis NIES-48 (E. gracilis) cells as large target cells and GCIY-EGFP (GCIY) cells derived from a gastric cancer cell line as small target cells. In E. gracilis cells sorting, the throughput is 23 kHz with a 92.8% success rate, 95.8% purity, and 90.8% cell viability. In GCIY sorting, the throughput is 11 kHz with a 97.8% success rate, 98.9% purity, and 90.7% cell viability. These results confirm that the proposed method sorts differently sized cells with high throughput and hence, overcomes the throughput-size trade-off that exists in conventional on-chip cell sorters.
- Published
- 2017
- Full Text
- View/download PDF
242. Improvement of the Measurement Range and Temperature Characteristics of a Load Sensor Using a Quartz Crystal Resonator with All Crystal Layer Components.
- Author
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Murozaki Y, Sakuma S, and Arai F
- Subjects
- Temperature, Quartz
- Abstract
Monitoring multiple biosignals, such as heart rate, respiration cycle, and weight transitions, contributes to the health management of individuals. Specifically, it is possible to measure multiple biosignals using load information obtained through contact with the environment, such as a chair and bed, in daily use. A wide-range load sensor is essential since load information contains multiple biosignals with various load ranges. In this study, a load sensor is presented by using a quartz crystal resonator (QCR) with a wide measurement range of 1.5 × 10⁶ (0.4 mN to 600 N), and its temperature characteristic of load is improved to -7 Hz/°C (-18 mN/°C). In order to improve the measurement range of the load, a design method of this sensor is proposed by restraining the buckling of QCR and by using a thinner QCR. The proposed sensor allows a higher allowable load with high sensitivity. The load sensor mainly consists of three layers, namely a QCR layer and two holding layers. As opposed to the conventional holding layer composed of silicon, quartz crystal is utilized for the holding layers to improve the temperature characteristic of the load sensor. In the study, multiple biosignals, such as weight and pulse, are detected by using a fabricated sensor.
- Published
- 2017
- Full Text
- View/download PDF
243. Arterial graft with elastic layer structure grown from cells.
- Author
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Yokoyama U, Tonooka Y, Koretake R, Akimoto T, Gonda Y, Saito J, Umemura M, Fujita T, Sakuma S, Arai F, Kaneko M, and Ishikawa Y
- Subjects
- Animals, Blood Vessel Prosthesis, Blood Vessel Prosthesis Implantation methods, Cells, Cultured, Elasticity, Humans, Hydrostatic Pressure, Rats, Stress, Mechanical, Tensile Strength, Muscle, Smooth, Vascular cytology, Myocytes, Smooth Muscle cytology, Tissue Engineering methods
- Abstract
Shortage of autologous blood vessel sources and disadvantages of synthetic grafts have increased interest in the development of tissue-engineered vascular grafts. However, tunica media, which comprises layered elastic laminae, largely determines arterial elasticity, and is difficult to synthesize. Here, we describe a method for fabrication of arterial grafts with elastic layer structure from cultured human vascular SMCs by periodic exposure to extremely high hydrostatic pressure (HP) during repeated cell seeding. Repeated slow cycles (0.002 Hz) between 110 and 180 kPa increased stress-fiber polymerization and fibronectin fibrillogenesis on SMCs, which is required for elastic fiber formation. To fabricate arterial grafts, seeding of rat vascular SMCs and exposure to the periodic HP were repeated alternatively ten times. The obtained medial grafts were highly elastic and tensile rupture strength was 1451 ± 159 mmHg, in which elastic fibers were abundantly formed. The patch medial grafts were sutured at the rat aorta and found to be completely patent and endothelialized after 2.5 months, although tubular medial constructs implanted in rats as interpositional aortic grafts withstood arterial blood pressure only in early acute phase. This novel organized self-assembly method would enable mass production of scaffold-free arterial grafts in vitro and have potential therapeutic applications for cardiovascular diseases.
- Published
- 2017
- Full Text
- View/download PDF
244. Mechanical diagnosis of human erythrocytes by ultra-high speed manipulation unraveled critical time window for global cytoskeletal remodeling.
- Author
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Ito H, Murakami R, Sakuma S, Tsai CD, Gutsmann T, Brandenburg K, Pöschl JM, Arai F, Kaneko M, and Tanaka M
- Subjects
- Adenosine Triphosphate metabolism, Cytoskeleton metabolism, Erythrocytes physiology, Humans, Microfluidics instrumentation, Time Factors, Cell Shape, Erythrocytes cytology, Microfluidics methods
- Abstract
Large deformability of erythrocytes in microvasculature is a prerequisite to realize smooth circulation. We develop a novel tool for the three-step "Catch-Load-Launch" manipulation of a human erythrocyte based on an ultra-high speed position control by a microfluidic "robotic pump". Quantification of the erythrocyte shape recovery as a function of loading time uncovered the critical time window for the transition between fast and slow recoveries. The comparison with erythrocytes under depletion of adenosine triphosphate revealed that the cytoskeletal remodeling over a whole cell occurs in 3 orders of magnitude longer timescale than the local dissociation-reassociation of a single spectrin node. Finally, we modeled septic conditions by incubating erythrocytes with endotoxin, and found that the exposure to endotoxin results in a significant delay in the characteristic transition time for cytoskeletal remodeling. The high speed manipulation of erythrocytes with a robotic pump technique allows for high throughput mechanical diagnosis of blood-related diseases.
- Published
- 2017
- Full Text
- View/download PDF
245. Temporal Transition of Mechanical Characteristics of HUVEC/MSC Spheroids Using a Microfluidic Chip with Force Sensor Probes.
- Author
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Ito K, Sakuma S, Kimura M, Takebe T, Kaneko M, and Arai F
- Abstract
In this paper, we focus on the mechanical characterization of co-cultured spheroids of human umbilical vein endothelial cells (HUVECs) and mesenchymal stem cells (MSC) (HUVEC/MSC spheroids). HUVEC/MSC spheroids aggregate during culture, thereby decreasing in size. Since this size decrease can be caused by the contractility generated by the actomyosin of MSCs, which are intracellular frames, we can expect that there is a temporal transition for the mechanical characteristics, such as stiffness, during culture. To measure the mechanical characteristics, we use a microfluidic chip that is integrated with force sensor probes. We show the details of the measurement configuration and the results of mechanical characterization of the HUVEC/MSC spheroids. To evaluate the stiffness of the spheroids, we introduce the stiffness index, which essentially shows a spring constant per unit size of the spheroid at a certain time during measurement. From the measurement results, we confirmed that the stiffness index firstly increased during the days of culture, although after four days of culture, the stiffness index decreased. We confirmed that the proposed system can measure the stiffness of HUVEC/MSC spheroids.
- Published
- 2016
- Full Text
- View/download PDF
246. Distinct patterns of cell motion inside a micro-channel under different osmotic conditions.
- Author
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Tsai CH, Kaneko M, Sakuma S, and Arai F
- Subjects
- Biomechanical Phenomena drug effects, Cell Shape drug effects, Dimethylpolysiloxanes chemistry, Erythrocytes cytology, Erythrocytes drug effects, Humans, Hypotonic Solutions pharmacology, Osmolar Concentration, Sodium Chloride pharmacology, Time Factors, Cell Movement drug effects, Microfluidics, Osmosis drug effects
- Abstract
The effect of osmotic condition on a living cell inside a micro-channel is firstly studied in this work. By utilizing a high-speed camera, we observed distinct patterns of cell motion under different osmotic conditions, which are established by saline with different concentrations of sodium chloride (NaCl). The cell motions are tracked by a computer, and are presented by the coordinates of location and time (x-t chart). The motions of cells under hypotonic condition (NaCl% < 0.9%) are convex curves on the chart while the ones under isotonic and hypertonic conditions (NaCl% ≥ 0.9%) are concave curves. Since saline is widely used in both medical practices and cell-related researches, our results point out two important facts: 1) Cells are sensitive to the percentage of NaCl. One percent difference in overall concentration makes dramatic changes in cell characteristics, such as cell stiffness. 2) The micro-channel method can clearly tell the difference between hypotonic, isotonic and hypertonic conditions according to the pattern of cell motion. Interpretations of the phenomena from different perspectives are also discussed in this paper.
- Published
- 2013
- Full Text
- View/download PDF
247. Powerful actuation of magnetized microtools by focused magnetic field for particle sorting in a chip.
- Author
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Yamanishi Y, Sakuma S, Onda K, and Arai F
- Subjects
- Finite Element Analysis, Image Processing, Computer-Assisted, Microtechnology, Magnetics, Microchip Analytical Procedures methods
- Abstract
This paper describes a novel powerful noncontact actuation of a magnetically driven microtool (MMT), achieved by magnetization of the MMT and focusing of the magnetic field in a microfluidic chip for particle sorting. The following are the highlights of this study: (1) an MMT was successfully fabricated from a mixture of neodymium powder and polydimethylsiloxane; the MMT was magnetized such that it acted as an elastic micromagnet with a magnetic flux density that increased by about 100 times after magnetization, and (2) a pair of sharp magnetic needles was fabricated adjacent to a microchannel in a chip by electroplating, in order to focus the magnetic flux density generated by the electromagnetic coils below the biochip; these needles contribute to miniaturization of an actuation module that would enable the integration of multiple functions in the limited area of a chip. FEM analysis of the magnetic flux density around the MMT showed that the magnetic flux density in the setup with the magnetic needles was around 8 times better than that in the setup without the needles. By magnetization, the drive frequency of the MMT improved by about 10 times--from 18 Hz to 180 Hz. We successfully demonstrated the separation of copolymer beads of a particular size in a chip by image sensing.
- Published
- 2010
- Full Text
- View/download PDF
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