Tohoku University View Institution's Website 17 articles published in JoVE Behavior Four Temporary Waterslide Designs Adapted to Different Slope Conditions to Encourage Child Socialization in Playgrounds Zhudi Hua1, Ting Tao1, Risa Akita1, Tomofusa Akita1, Yoshiaki Hayakawa1, Masanori Hariyama2, Hayato Sakurai3, Ricki Colman4, Mamiko Koshiba1,2,3 1Graduate School of Sciences and Technology for Innovation, Yamaguchi University, 2Graduate School of Information Sciences, Tohoku University, 3Pediatrics, Saitama Medical University Hospital, 4Department of Cell and Regenerative Biology, University of Wisconsin-Madison Early life social learning is enhanced by interactions with effectively designed environments. Four events were held at different city parks using inexpensive, temporary waterslides to stimulate social learning. This study describes the prototypes used and the evaluation of the children's interactions. Developmental Biology Fluorescent In Situ Hybridization and 5-Ethynyl-2'-Deoxyuridine Labeling for Stem-Like Cells in the Hydrozoan Jellyfish Cladonema pacificum Sosuke Fujita1, Erina Kuranaga1, Masayuki Miura2, Yu-ichiro Nakajima2 1Graduate School of Life Sciences, Tohoku University, 2Graduate School of Pharmaceutical Sciences, The University of Tokyo Here, we describe a protocol for visualizing stem-like proliferating cells in the jellyfish Cladonema. Whole-mount fluorescent in situ hybridization with a stem cell marker allows for the detection of stem-like cells, and 5-ethynyl-2'-deoxyuridine labeling enables the identification of proliferating cells. Together, actively proliferating stem-like cells can be detected. Chemistry Self-Assembly of Hybrid Lipid Membranes Doped with Hydrophobic Organic Molecules at the Water/Air Interface Xingyao Feng1, Teng Ma2, Daisuke Tadaki1, Ayumi Hirano-Iwata1,2 1Research Institute of Electrical Communication, Tohoku University, 2Advanced Institute for Materials Research, Tohoku University We report a protocol for producing a hybrid lipid membrane at the water/air interface by doping the lipid bilayer with copper (II) 2,9,16,23-tetra-tert-butyl-29H,31H-phthalocyanine (CuPc) molecules. The resulting hybrid lipid membrane has a lipid/CuPc/lipid sandwich structure. This protocol can also be applied to the formation of other functional nanomaterials. Immunology and Infection Application of Consistent Massage-Like Perturbations on Mouse Calves and Monitoring the Resulting Intramuscular Pressure Changes Naoyoshi Sakitani*1, Takahiro Maekawa*1, Kumiko Saitou1,2, Katsuhiko Suzuki3, Shuhei Murase1,4, Masakuni Tokunaga1, Daisuke Yoshino5, Keisuke Sawada6, Atsushi Takashima7, Motoshi Nagao1, Toru Ogata1, Yasuhiro Sawada1,8 1Department of Rehabilitation for Motor Functions, National Rehabilitation Center for Persons with Disabilities, 2Graduate School of Sport Sciences, Waseda University, 3Faculty of Sport Sciences, Waseda University, 4Department of Orthopaedic Surgery, Graduate School of Medicine, The University of Tokyo, 5Frontier Research Institute for Interdisciplinary Sciences, Tohoku University, 6University of Cincinnati College of Medicine, 7Department of Assistive Technology, National Rehabilitation Center for Persons with Disabilities, 8Department of Clinical Research, National Rehabilitation Center for Persons with Disabilities Here we describe the protocols for applying defined mechanical loads to mouse calves and for monitoring the concomitant intramuscular pressure changes. The experimental systems that we have developed can be useful for investigating the mechanism behind the beneficial effects of physical exercise and massage. Bioengineering Evaluation of Photosynthetic Behaviors by Simultaneous Measurements of Leaf Reflectance and Chlorophyll Fluorescence Analyses Kaori Kohzuma1 1Graduate School of Life Sciences, Tohoku University We describe a new technical approach to study photosynthetic responses in higher plants involving simultaneous measurements of chlorophyll a fluorescence and leaf reflectance using a PAM and a spectral radiometer for the detection of signals from the same leaf area in Arabidopsis. Immunology and Infection Identification of Mouse and Human Antibody Repertoires by Next-Generation Sequencing Lin Sun1, Naoko Kono2, Hiroyuki Toh3, Hanbing Xue1, Kaori Sano4,5, Tadaki Suzuki4, Akira Ainai4, Yasuko Orba6, Junya Yamagishi7,8, Hideki Hasegawa4,5, Yoshimasa Takahashi9, Shigeyuki Itamura2, Kazuo Ohnishi9,10 1Graduate School of Life and Environmental Sciences, University of Tsukuba, 2Center for Influenza Virus Research, National Institute of Infectious Diseases, 3School of Science and Technology, Kwansei Gakuin University, 4Department of Pathology, National Institute of Infectious Diseases, 5Division of Infectious Diseases Pathology, Department of Global Infectious Diseases, Tohoku University Graduate School of Medicine, 6Division of Molecular Pathobiology, Research Center for Zoonosis Control, Hokkaido University, 7Division of Collaboration and Education, Research Center for Zoonosis Control, Hokkaido University, 8Global Station for Zoonosis Control, GI-CoRE, Hokkaido University, 9Department of Immunology, National Institute of Infectious Diseases, 10Faculty of Life and Environmental Sciences, University of Tsukuba Here, we describe protocols for the analysis and visualization of the structure and constitution of whole antibody repertoires. This involves the acquisition of vast sequences of antibody RNA using next-generation sequencing. Immunology and Infection Assays for the Specific Growth Rate and Cell-binding Ability of Rotavirus Syun-suke Kadoya1, Daisuke Sano1,2 1Department of Civil and Environmental Engineering, Tohoku University, 2Department of Frontier Science for Advanced Environment, Graduate School of Environmental Studies, Tohoku University Here we present two protocols, one for measuring the specific growth rate and the other for the cell-binding ability of rotavirus using the plaque assay and RT-qPCR. These protocols are available for confirming the differences in phenotypes between rotavirus strains. Biology Imaging FITC-dextran as a Reporter for Regulated Exocytosis Ofir Klein1, Amit Roded1, Koret Hirschberg2, Mitsunori Fukuda3, Stephen J. Galli4, Ronit Sagi-Eisenberg1 1Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, 2Department of Pathology, Sackler Faculty of Medicine, Tel Aviv University, 3Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University, 4Departments of Pathology and of Microbiology and Immunology and Sean N. Parker Center for Allergy and Asthma Research, School of Medicine, Stanford University Here we detail a method for live cell imaging of regulated exocytosis. This method utilizes FITC-dextran, which accumulates in lysosome-related organelles, as a reporter. This simple method also allows distinguishing between different modes of regulated exocytosis in cells that are difficult to manipulate genetically. Bioengineering Microhoneycomb Monoliths Prepared by the Unidirectional Freeze-drying of Cellulose Nanofiber Based Sols: Method and Extensions Zheng-Ze Pan1,2, Hirotomo Nishihara3, Wei Lv1, Cong Wang1,2, Yi Luo1,2, Liubing Dong1,2, Houfu Song1,4, Wenjie Zhang2, Feiyu Kang1,2,4, Takashi Kyotani3, Quan-Hong Yang1,4,5 1Engineering Laboratory for Functionalized Carbon Materials and Shenzhen Key Laboratory for Graphene-based Materials, Graduate School at Shenzhen, Tsinghua University, 2School of Materials Science and Engineering, Tsinghua University, 3Institute of Multidisciplinary Research for Advanced Materials, Tohoku University, 4Tsinghua-Berkeley Shenzhen Institute (TBSI), Tsinghua University, 5School of Chemical Engineering and Technology, Tianjin University Here, we present a general protocol to prepare a variety of microhoneycomb monoliths (MHMs) in which fluid can pass through with an extremely low pressure drop. MHMs obtained are expected to be used as filters, catalyst supports, flow-type electrodes, sensors and scaffolds for biomaterials. Engineering Growth and Electrostatic/chemical Properties of Metal/LaAlO3/SrTiO3 Heterostructures Diogo Castro Vaz1, Edouard Lesne1,2, Anke Sander1, Hiroshi Naganuma1,3, Eric Jacquet1, Jacobo Santamaria1,4, Agnès Barthélémy1, Manuel Bibes1 1Unité Mixte de Physique CNRS/Thales, Université Paris-Saclay, 2Max Planck Institut für Mikrostrukturphysik, 3Department of Applied Physics, Tohoku University, 4Instituto de Magnetismo Aplicado, Universidad Complutense de Madrid We fabricate metal/LaAlO3/SrTiO3 heterostructures using a combination of pulsed laser deposition and in situ magnetron sputtering. Through magnetotransport and in situ X-ray photoelectron spectroscopy experiments, we investigate the interplay between electrostatic and chemical phenomena of the quasi two-dimensional electron gas formed in this system. Developmental Biology Immobilization of Caenorhabditis elegans to Analyze Intracellular Transport in Neurons Shinsuke Niwa1 1Frontier Research Institute for Interdisciplinary Sciences and Graduate School of Life Sciences, Tohoku University Caenorhabditis elegans (C. elegans) is a good model to study axonal and intracellular transport. Here, I describe a protocol for in vivo recording and analysis of axonal and intraflagellar transport in C. elegans. Medicine Isolation and Profiling of MicroRNA-containing Exosomes from Human Bile Ling Li1, Klaus B. Piontek1, Vivek Kumbhari1, Masaharu Ishida2, Florin M. Selaru1 1Division of Gastroenterology and Hepatology, Department of Medicine, Johns Hopkins University School of Medicine, 2Department of Surgery, Tohoku University Bile fluid is a valuable source of extracellular vesicles/exosomes that contain potentially important biomarkers. This protocol represents a robust method to isolate exosomes from human bile for further analyses including miRNA profiling. Bioengineering Photopatterning Proteins and Cells in Aqueous Environment Using TiO2 Photocatalysis Hideaki Yamamoto1,2, Takanori Demura3, Kohei Sekine3, Sho Kono3, Michio Niwano2,4, Ayumi Hirano-Iwata2,5, Takashi Tanii6 1Frontier Research Institute for Interdisciplinary Sciences, Tohoku University, 2CREST, Japan Science and Technology Agency, 3School of Fundamental Science and Engineering, Waseda University, 4Research Institute of Electrical Communication, Tohoku University, 5Graduate School of Biomedical Engineering, Tohoku University, 6Faculty of Science and Engineering, Waseda University We describe a protocol for modifying cell affinity of a scaffold surface in aqueous environment. The method takes advantage of titanium dioxide photocatalysis to decompose organic film in the photo-irradiated region. We show that it can be used to create microdomains of scaffolding proteins, both ex situ and in situ. Immunology and Infection Investigating Mast Cell Secretory Granules; from Biosynthesis to Exocytosis Nurit P. Azouz1,2, Mitsunori Fukuda3, Marc E. Rothenberg2, Ronit Sagi-Eisenberg1 1Department of Cell and Developmental Biology, Sackler Faculty of Medicine, Tel Aviv University, 2Division of Allergy and Immunology, Department of Pediatrics, Cincinnati Children's Hospital Medical Center, University of Cincinnati College of Medicine, 3Laboratory of Membrane Trafficking Mechanisms, Department of Developmental Biology and Neurosciences, Graduate School of Life Sciences, Tohoku University The goal of the present protocol was to develop a method that will allow functional genomic analyses of mast cell secretion. The protocol is based on quantitative assessment of the release of a fluorescent reporter gene cotrasfected with the gene of interest and real time analyses of the secretory granule's morphology. Biology Preparation of Rat Serum Suitable for Mammalian Whole Embryo Culture Masanori Takahashi1,2, Sayaka Makino3, Takako Kikkawa3, Noriko Osumi3 1Graduate School of Medicine, Jichi Medical University, 2Division of Biology, Center for Molecular Medicine, Jichi Medical University, 3Department of Developmental Neuroscience, United Center for Advanced Research and Translational Medicine (ART), Tohoku University School of Medicine Mammalian whole embryo culture (WEC) is widely used in teratology and developmental biology. Immediately centrifuged rat serum is commonly provided as a medium for both mouse and rat WEC. In this video, we demonstrate our standard protocol for the preparation of high-quality rat serum suitable for mammalian WEC. Biology Identifying the Effects of BRCA1 Mutations on Homologous Recombination using Cells that Express Endogenous Wild-type BRCA1 Jeffrey Parvin1, Natsuko Chiba2, Derek Ransburgh1 1Department of Biomedical Informatics, The Ohio State University, 2Departments of Molecular Immunology and Clinical Oncology, Tohoku University We provide a method for testing BRCA1 variants in a tissue culture based assay for homologous recombination repair of DNA damage by depleting endogenous BRCA1 protein from a cell using RNAi and replacing it with a BRCA1 point mutant that contains a coding change. Biology The Method of Rodent Whole Embryo Culture using the Rotator-type Bottle Culture System Masanori Takahashi1, Noriko Osumi1,2 1Division of Developmental Neuroscience, United Centers for Advanced Research and Translational Medicine (ART), Tohoku University Graduate School of Medicine, 2The Core Research for Evolutional Science and Technology (CREST), Japan Science and Technology Corporation (JST) Whole embryo culture technique allows us to culture mouse and rat embryos ex vivo condition during limited periods corresponding to midgestation stages. In this video protocol, we demonstrate our standard procedures of rat whole embryo culture after E12.5 using the rotator-type bottle culture system.