Tokyo Institute of Technology 14 articles published in JoVE Biology Reactivation of Demembranated Cell Models in Chlamydomonas reinhardtii Noriko Ueki1, Atsuko Isu2, Ken‒ichi Wakabayashi2,3 1Science Research Center, Hosei University, 2Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, 3School of Life Science and Technology, Tokyo Institute of Technology The in vitro reactivation of motile cells is a crucial experiment in understanding the mechanisms of cell motility. The protocol describes reactivating the demembranated cell models of Chlamydomonas reinhardtii, a model organism to study cilia/flagella. Biology Observation of Photobehavior in Chlamydomonas reinhardtii Noriko Ueki*1, Atsuko Isu*2, Ayaka Kyuji*2,3, Yuma Asahina*2,3, Satoaki So*2,3, Rina Takahashi*2,3, Toru Hisabori2,3, Ken‒ichi Wakabayashi2,3 1Science Research Center, Hosei University, 2Laboratory for Chemistry and Life Science, Institute of Innovative Research, Tokyo Institute of Technology, 3School of Life Science and Technology, Tokyo Institute of Technology Most swimming photoautotrophic organisms show photo-induced behavioral changes (photobehavior). The present protocol observes the said photobehavior in the model organism Chlamydomonas reinhardtii. Bioengineering Automated Microbial Cultivation and Adaptive Evolution using Microbial Microdroplet Culture System (MMC) Xingjin Jian*1,2, Xiaojie Guo*1,2,3, Jia Wang4, Zheng Lin Tan1,2,6, Xin-hui Xing1,2,5, Liyan Wang3, Chong Zhang1,2,5 1Department of Chemical Engineering, Institute of Biochemical Engineering, Tsinghua University, 2Key Laboratory of Industrial Biocatalysis, Ministry of Education, Tsinghua University, 3Luoyang TMAXTREE Biotechnology Co., Ltd., 4 This protocol describes how to use the Microbial Microdroplet Culture system (MMC) to conduct automated microbial cultivation and adaptive evolution. MMC can cultivate and sub-cultivate microorganisms automatically and continuously and monitor online their growth with relatively high throughput and good parallelization, reducing labor and reagent consumption. Biology Bead Loading Proteins and Nucleic Acids into Adherent Human Cells Charlotte Ayn Cialek1, Gabriel Galindo1, Amanda Lynn Koch1, Matthew Neeley Saxton1, Timothy John Stasevich1,2 1Department of Biochemistry and Molecular Biology, Colorado State University, 2World Research Hub Initiative, Institute of Innovative Research, Tokyo Institute of Technology Bead loading introduces proteins, plasmids, and particles into adherent mammalian cells. This cell loading technique is inexpensive, rapid, and does not substantially affect cell health. It is best suited for live-cell imaging. Biochemistry Real-time Observation of the DNA Strand Exchange Reaction Mediated by Rad51 Kentaro Ito1, Bilge Argunhan1, Hideo Tsubouchi1, Hiroshi Iwasaki1 1Cell Biology Center, Institute of Innovative Research, Tokyo Institute of Technology Fluorescence resonance energy transfer-based real-time observation systems of the DNA strand exchange reaction mediated by Rad51 were developed. Using the protocols presented here, we are able to detect the formation of reaction intermediates and their conversion into products, while also analyzing the enzymatic kinetics of the reaction. Immunology and Infection Enhancement Method of Surface Acoustic Wave-Atomizer Efficiency for Olfactory Display Takamichi Nakamoto1, Sami Ollila1, Shingo Kato1, Haining Li1,2, Guiping Qi1 1Tokyo Institute of Technology, 2University of Electronic Science and Technology of China We establish here a method for coating the surface of a surface acoustic wave (SAW) device with amorphous Teflon film to improve the atomization efficiency required for application to an olfactory display. Bioengineering Fabrication of a Multiplexed Artificial Cellular MicroEnvironment Array Yasumasa Mashimo1,2, Momoko Yoshioka1, Yumie Tokunaga1, Christopher Fockenberg1, Shiho Terada1, Yoshie Koyama1, Teiko Shibata-Seki2, Koki Yoshimoto1, Risako Sakai1, Hayase Hakariya1, Li Liu1, Toshihiro Akaike3, Eiry Kobatake2, Siew-Eng How4, Motonari Uesugi1,5, Yong Chen1,6, Ken-ichiro Kamei1 1Institute for Integrated Cell-Material Sciences (WPI-iCeMS), Kyoto University, 2Department of Life Science and Technology, School of Life Science and Technology, Tokyo Institute of Technology, 3Biomaterials Center for Regenerative Medical Engineering, Foundation for Advancement of International Science, 4Faculty of Science and Natural Resources, Universiti Malaysia Sabah, 5Institute for Chemical Research, Kyoto University, 6Ecole Normale Supérieure This article describes the detailed methodology to prepare a Multiplexed Artificial Cellular MicroEnvironment (MACME) array for high-throughput manipulation of physical and chemical cues mimicking in vivo cellular microenvironments and to identify the optimal cellular environment for human pluripotent stem cells (hPSCs) with single-cell profiling. Chemistry Preparation of Polyoxometalate-based Photo-responsive Membranes for the Photo-activation of Manganese Oxide Catalysts Akira Yamaguchi1,5, Toshihiro Takashima2, Kazuhito Hashimoto1,6, Ryuhei Nakamura3,4 1Department of Applied Chemistry, The University of Tokyo, 2Clean Energy Research Center, University of Yamanashi, 3Biofunctional Catalyst Research Team, RIKEN Center for Sustainable Resource Science, 4Earth-Life Science Institute (ELSI), Tokyo Institute of Technology, 5Department of Materials Science and Engineering, Tokyo Institute of Technology, 6National Institute for Materials Science Here, we present a protocol to prepare charge transfer chromophores based on a polyoxometalate/polymer composite membrane. Neuroscience Analyzing Synaptic Modulation of Drosophila melanogaster Photoreceptors after Exposure to Prolonged Light Atsushi Sugie1,2,5, Christoph Möhl3, Satoko Hakeda-Suzuki4, Hideaki Matsui1,2, Takashi Suzuki*4, Gaia Tavosanis*5 1Department of Neuroscience of Disease, Center for Transdisciplinary Research, Niigata University, 2Brain Research Institute, Niigata University, 3Image and Data Analysis Facility, German Center for Neurodegenerative Diseases (DZNE), 4Graduate School of Life Science and Technology, Tokyo Institute of Technology (Titech), 5Dendrite Differentiation, German Center for Neurodegenerative Diseases (DZNE) Here we show how to quantify the number and spatial distribution of synaptic active zones in Drosophila melanogaster photoreceptors, highlighted with a genetically encoded molecular marker, and their modulation after prolonged exposure to light. Chemistry Direct Imaging of Laser-driven Ultrafast Molecular Rotation Kenta Mizuse1, Romu Fujimoto1, Nobuo Mizutani2, Yasuhiro Ohshima1,3 1Department of Chemistry, Tokyo Institute of Technology, 2Equipment Development Center, Institute for Molecular Science, 3Department of Photo-molecular Science, Institute for Molecular Science We present a protocol for creating a real-time movie of a molecular rotational wave packet using a high-resolution Coulomb explosion imaging setup. Behavior Visualization Method for Proprioceptive Drift on a 2D Plane Using Support Vector Machine Daisuke Tajima1, Tota Mizuno2, Yuichiro Kume3, Takako Yoshida1 1Applied Brain Science Laboratory, Department of Mechanical Sciences and Engineering, Tokyo Institute of Technology, 2Department of Informatics, Graduate School of Informatics and Engineering, The University of Electro-Communications, 3Department of Media and Image Technology, Faculty of Engineering, Tokyo Polytechnic University This article describes a novel method to estimate proprioceptive drift on a 2D plane using the mirror illusion and combining a psychophysical procedure with an analysis using machine learning. Chemistry Synthesis of Cyclic Polymers and Characterization of Their Diffusive Motion in the Melt State at the Single Molecule Level Satoshi Habuchi1, Takuya Yamamoto2,3, Yasuyuki Tezuka3 1Biological and Environmental Sciences and Engineering Division, King Abdullah University of Science and Technology (KAUST), 2Division of Applied Chemistry, Faculty of Engineering, Hokkaido University, 3Department of Organic and Polymeric Materials, Tokyo Institute of Technology A protocol for the synthesis and characterization of diffusive motion of cyclic polymers at the single molecule level is presented. Engineering Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets Masamune Morita1, Hitoyoshi Yamashita1,2, Masayuki Hayakawa1, Hiroaki Onoe3, Masahiro Takinoue1,4 1Department of Computational Intelligence and Systems Science, Interdisciplinary Graduate School of Science and Engineering, Tokyo Institute of Technology, 2Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, 3Department of Mechanical Engineering, Keio University, 4PRESTO, Japan Science and Technology Agency Here, we demonstrate a simple production method for size-controllable, monodisperse, water-in-oil (W/O) microdroplets using a capillary-based centrifugal microfluidic device. This method requires only a small sample volume and enables high-yield production. We expect this method will be useful for rapid biochemical and cellular analyses. Chemistry Conducting Miller-Urey Experiments Eric T. Parker1, James H. Cleaves2,3, Aaron S. Burton4, Daniel P. Glavin5, Jason P. Dworkin5, Manshui Zhou1, Jeffrey L. Bada6, Facundo M. Fernández1 1School of Chemistry and Biochemistry, Georgia Institute of Technology, 2Earth-Life Science Institute, Tokyo Institute of Technology, 3Institute for Advanced Study, 4Astromaterials Research and Exploration Science Directorate, NASA Johnson Space Center, 5Goddard Center for Astrobiology, NASA Goddard Space Flight Center, 6Geosciences Research Division, Scripps Institution of Oceanography, University of California at San Diego The Miller-Urey experiment was a pioneering study regarding the abiotic synthesis of organic compounds with possible relevance to the origins of life. Simple gases were introduced into a glass apparatus and subjected to an electric discharge, simulating the effects of lightning in the primordial Earth’s atmosphere-ocean system. The experiment was conducted for one week, after which, the samples collected from it were analyzed for the chemical building blocks of life.
