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Keio University

7 ARTICLES PUBLISHED IN JoVE

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Medicine

Visualization and Analysis of Blood Flow and Oxygen Consumption in Hepatic Microcirculation: Application to an Acute Hepatitis Model
Kosuke Tsukada 1, Makoto Suematsu 2,3
1Department of Applied Physics and Physico-Informatics, Keio University, 2Department of Biochemistry, School of Medicine, Keio University, 3Exploratory Research for Advanced Technology (ERATO), Suematsu Gas Biology Project, Japan Science and Technology Agency (JST)

An optical system was developed to visualize hepatic microcirculation with FITC-labeled erythrocytes and to measure the partial pressure of oxygen in the microvessels with laser-assisted phosphorimetry. This method can be used to investigate physiological and pathological mechanisms by analyzing microvascular structure, diameter, blood flow velocity, and oxygen tension.

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Biology

The Green Monster Process for the Generation of Yeast Strains Carrying Multiple Gene Deletions
Yo Suzuki 1, Jason Stam 1, Mark Novotny 2, Nozomu Yachie 3, Roger S. Lasken 2, Frederick P. Roth 3,4
1Department of Synthetic Biology and Bioenergy, J. Craig Venter Institute, 2Department of Microbial and Environmental Genomics, J. Craig Venter Institute, 3Donnelly Centre & Department of Molecular Genetics, University of Toronto, 4Lunenfeld Research Institute, Mt Sinai Hospital

The Green Monster method enables the rapid assembly of multiple deletions marked with a reporter gene encoding green fluorescent protein. This method is based on driving yeast strains through repeated cycles of sexual assortment of deletions and fluorescence-based enrichment of cells carrying more deletions.

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Engineering

Capillary-based Centrifugal Microfluidic Device for Size-controllable Formation of Monodisperse Microdroplets
Masamune Morita 1, Hitoyoshi Yamashita 1,2, Masayuki Hayakawa 1, Hiroaki Onoe 3, Masahiro Takinoue 1,4
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.

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Cancer Research

Induction and Diagnosis of Tumors in Drosophila Imaginal Disc Epithelia
Kenta Morimoto 1,2, Yoichiro Tamori 1
1Structural Biology Center, National Institute of Genetics and Department of Genetics, School of Life Science, SOKENDAI, 2Graduate School of Media and Governance, Keio University

Mosaic clone analysis in Drosophila imaginal disc epithelia is a powerful model system to study the genetic and cellular mechanisms of tumorigenesis. Here we describe a protocol to induce tumors in Drosophila wing imaginal discs using the GAL4-UAS system, and introduce a diagnosis method to classify the tumor phenotypes.

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Behavior

A Method to Study Adaptation to Left-Right Reversed Audition
Atsushi Aoyama 1,2
1Faculty of Environment and Information Studies, Shonan Fujisawa Campus (SFC), Keio University, 2School of Information Environment, Tokyo Denki University

The present study proposes a protocol to investigate the adaptation to left-right reversed audition achieved only by wearable devices, using neuroimaging, which can be an effective tool for uncovering the adaptability of humans to a novel environment in the auditory domain.

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Genetics

Ultralow Input Genome Sequencing Library Preparation from a Single Tardigrade Specimen
Yuki Yoshida 1,2, Sayuri Konno 1,3, Ryousuke Nishino 1,3, Yumi Murai 1,3, Masaru Tomita 1,2,3, Kazuharu Arakawa 1,2,3
1Institute for Advanced Biosciences, Keio University, 2Systems Biology Program, Graduate School of Media and Governance, Keio University, 3Faculty of Environment and Information Studies, Keio University

Contamination during the genomic sequencing of microscopic organisms remains a large problem. Here, we show a method to sequence the genome of a tardigrade from a single specimen with as little as 50 pg of genomic DNA without whole genome amplification to minimize the risk of contamination.

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Genetics

Use of Freeze-thawed Embryos for High-efficiency Production of Genetically Modified Mice
Hirofumi Nishizono *1,2,3, Mohamed Darwish *4,5, Hideki Uosaki 6,7, Nanami Masuyama 8,9,10, Motoaki Seki 8,11, Hiroyuki Abe 3, Nozomu Yachie 8,9,10,12,13, Ryohei Yasuda 1
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.

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