Chiba University 6 articles published in JoVE Biology Rapid Freezing using Sandwich Freezing Device for Good Ultrastructural Preservation of Biological Specimens in Electron Microscopy Masashi Yamaguchi1, Azusa Takahashi-Nakaguchi1, Katsuyuki Uematsu2, Masaki Taguchi2, Michiyo Sato-Okamoto1, Hiroji Chibana1 1Medical Mycology Research Center, Chiba University, 2Marine Works Japan Ltd. Here we show how to use the sandwich freezing device for rapid freezing of biological specimens, including bacteria, yeast, cultured cells, isolated cells, animal and human tissues, and viruses. We also show how to prepare specimens for ultrathin sectioning after rapid freezing. Genetics Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice Hirofumi Nishizono*1,2,3, Mohamed Darwish*4,5, Hideki Uosaki6,7, Nanami Masuyama8,9,10, Motoaki Seki8,11, Hiroyuki Abe3, Nozomu Yachie8,9,10,12,13, Ryohei Yasuda1 1Max Planck Florida Institute for Neuroscience, 2Life Science Research Center, University of Toyama, 3Department of Biochemical Engineering, Graduate School of Science and Engineering, Yamagata University, 4Graduate School of Innovative Life Science, University of Toyama, 5Department of Biochemistry, Faculty of Pharmacy, Cairo University, 6Division of Regenerative Medicine, Center for Molecular Medicine, Jichi Medical University, 7Division of Stem Cell Research and Drug Development, Center for Development of Advanced Medical Technology, Jichi Medical University, 8Synthetic Biology Division, Research Center for Advanced Science and Technology, University of Tokyo, 9Institute for Advanced Biosciences, Keio University, 10Graduate School of Media and Governance, Keio University, 11Department of Molecular Oncology, Graduate School of Medicine, Chiba University, 12Department of Biological Sciences, School of Science, University of Tokyo, 13College of Arts and Sciences, University of Tokyo Here, we present a modified method for cryopreservation of one-cell embryos as well as a protocol that couples the use of freeze-thawed embryos and electroporation for the efficient generation of genetically modified mice. Developmental Biology Evaluation of Fertilization State by Tracing Sperm Nuclear Morphology in Arabidopsis Double Fertilization Taro Takahashi1, Tomoko Igawa1 1Graduate School of Horticulture, Chiba University We demonstrate a method to determine successful or failed fertilization on the basis of sperm nuclear morphology in Arabidopsis double fertilization using an epifluorescence microscope. Medicine Semi-quantitative Assessment Using [18F]FDG Tracer in Patients with Severe Brain Injury Tomohiro Yamaki1,2, Shinji Onodera2, Tomoki Uchida2, Yoshihiro Ozaki2, Kazuaki Yokoyama3, Haruko Henmi2, Mizuho Kamezawa2, Miyoko Hayakawa2, Daisuke Itou1, Nobuo Oka1,2, Masaru Odaki1, Yasuo Iwadate4, Shigeki Kobayashi1 1 [18F]-fluorodeoxyglucose (FDG) positron emission tomography-computed tomography is useful for studying glucose metabolism related to brain function. Here, we present a protocol for an [18F]FDG tracer set-up and semiquantitative assessment of the region-of-interest analysis for targeted brain areas associated with clinical manifestations in patients with severe traumatic brain injury. Engineering A Method for Obtaining Serial Ultrathin Sections of Microorganisms in Transmission Electron Microscopy Masashi Yamaguchi1, Hiroji Chibana1 1Medical Mycology Research Center, Chiba University This study presents reliable and easy procedures for obtaining serial ultrathin sections of a microorganism without expensive equipment in transmission electron microscopy. Environment Field Experiments of Pollination Ecology: The Case of Lycoris sanguinea var. sanguinea Futa Yamaji1, Takeshi A. Ohsawa1 1Department of Biology, Chiba University To reveal the pollinator effectiveness of a given plant species, multiple methods of field experiments have been developed. This study demonstrates the basic methods of field experiments for pollination ecology using the case study of Lycoris sanguinea var. sanguinea and the novel pollination mechanism, breaking-bud pollination.
