Kumamoto University 6 articles published in JoVE Chemistry Magnetometric Characterization of Intermediates in the Solid-State Electrochemistry of Redox-Active Metal-Organic Frameworks Qi Chen1, Zhongyue Zhang2, Kunio Awaga1,3 1Department of Chemistry, Graduate School of Science, Nagoya University, 2International Research Organization for Advanced Science and Technology, Kumamoto University, 3Integrated Research Consortium on Chemical Sciences, Nagoya University Ex situ magnetic surveys can directly provide bulk and local information on a magnetic electrode to reveal its charge storage mechanism step by step. Herein, electron spin resonance (ESR) and magnetic susceptibility are demonstrated to monitor the evaluation of paramagnetic species and their concentration in a redox-active metal-organic framework (MOF). Medicine Sample Preparation for Computed Tomography-based Three-dimensional Visualization of Murine Hind-limb Vessels Daiki Seya1, Yuqing Xu2,3, Toshifumi Mukunoki4, Kenichi Tsujita3, Osamu Nakagawa1, Yuichiro Arima1,2,3,4 1Department of Molecular Physiology, National Cerebral and Cardiovascular Center Research Institute, 2International Research Center for Medical Sciences (IRCMS), Kumamoto University, 3Department of Cardiovascular Medicine, Graduate School of Medical Sciences, Kumamoto University, 4X-Earth Center, Faculty of Advanced Science and Technology, Kumamoto University Here, we describe a visualization and quantification method for murine hind-limb vessels using micro-X-ray computed tomography. Biology Isolation and Culture of Primary Oral Keratinocytes from the Adult Mouse Palate Yen Xuan Ngo1,2,3, Kenta Haga4, Ayako Suzuki4, Hiroko Kato4, Hiromi Yanagisawa1,5, Kenji Izumi4, Aiko Sada1,3 1Life Science Center for Survival Dynamics, Tsukuba Advanced Research Alliance (TARA), University of Tsukuba, 2Ph.D. Program in Human Biology, School of Integrative and Global Majors, University of Tsukuba, 3International Research Center for Medical Sciences (IRCMS), Kumamoto University, 4Division of Biomimetics, Faculty of Dentistry and Graduate School of Medical and Dental Sciences, Niigata University, 5Faculty of Medicine, University of Tsukuba The present protocol describes the isolation and culture of oral keratinocytes derived from the adult mouse palate. An evaluation method using immunostaining is also reported. Environment An Induction System for Clustered Stomata by Sugar Solution Immersion Treatment in Arabidopsis thaliana Seedlings Kae Akita1, Takumi Higaki2 1Department of Integrated Frontier Sciences, The University of Tokyo, 2International Research Organization for Advanced Science and Technology, Kumamoto University The goal of this protocol is to demonstrate how to induce clustered stomata in cotyledons of Arabidopsis thaliana seedlings by immersion treatment with a sugar-containing medium solution and how to observe intracellular structures such as chloroplasts and microtubules in the clustered guard cells using confocal laser microscopy. Neuroscience In Vivo Two-photon Imaging of Cortical Neurons in Neonatal Mice Hidenobu Mizuno1,2,3, Shingo Nakazawa2,3, Takuji Iwasato2,3 1International Research Center for Medical Sciences (IRCMS), Kumamoto University, 2Division of Neurogenetics, National Institute of Genetics, 3Department of Genetics, SOKENDAI (The Graduate University for Advanced Studies) We present an in vivo two-photon imaging protocol for imaging the cerebral cortex of neonatal mice. This method is suitable for analyzing the developmental dynamics of cortical neurons, the molecular mechanisms that control the neuronal dynamics, and the changes in neuronal dynamics in disease models. Bioengineering Perfusable Vascular Network with a Tissue Model in a Microfluidic Device Yuji Nashimoto1, Yukako Teraoka1, Ramin Banan Sadeghian1, Akiko Nakamasu2, Yuichiro Arima3, Sanshiro Hanada3, Hidetoshi Kotera1, Koichi Nishiyama3, Takashi Miura2, Ryuji Yokokawa1 1Department of Micro Engineering, Kyoto University, 2Graduate School of Medical Sciences, Kyushu University, 3International Research Center for Medical Sciences (IRCMS), Kumamoto University The protocol describes how to engineer a perfusable vascular network in a spheroid. The spheroid's surrounding microenvironment is devised to induce angiogenesis and connect the spheroid to the microchannels in a microfluidic device. The method allows the perfusion of the spheroid, which is a long-awaited technique in three-dimensional cultures.