Yonsei University College of Medicine 9 articles published in JoVE Medicine In vitro Time-lapse Live-Cell Imaging to Explore Cell Migration toward the Organ of Corti Jeong-Eun Park*1,2,3, Su Hoon Lee*1,2, Dong Jun Park1,2, Young Joon Seo1,2, Sung Kyun Kim3 1Department of Otorhinolaryngology, Yonsei University Wonju College of Medicine, 2Research Institute of Hearing Enhancement, Yonsei University Wonju College of Medicine, 3Department of Otorhinolaryngology Head and Neck Surgery, Hallym University College of Medicine In this study, we present a real-time imaging method using confocal microscopy to observe cells moving toward damaged tissue by ex vivo incubation with the cochlear epithelium containing the organ of Corti. Bioengineering Use of Micro X-ray Computed Tomography with Phosphotungstic Acid Preparation to Visualize Human Fibromuscular Tissue Jehoon O1, Hyun-Jin Kwon1, Shin Hyung Kim2, Tae-Hyeon Cho1, Hun-Mu Yang1,3 1Department of Anatomy, Yonsei University College of Medicine, 2Department of Anesthesiology and Pain Medicine, Anesthesia and Pain Research Institute, Yonsei University College of Medicine, 3Surgical Anatomy Education Centre at the Yonsei University College of Medicine Micro X-ray computed tomography is effective in obtaining three-dimensional information from undamaged human specimens but has limited success in observing soft tissues. The use of phosphotungstic acid contrast agent can resolve this issue. We implemented this contrast agent to examine human delicate fibromuscular tissues (the orbicularis retaining ligament). Medicine Porcine As a Training Module for Head and Neck Microvascular Reconstruction Mohammad Ali Alessa*1,2, Sang Hyun Kwak*2, Young Woo Lee2, Mi-Lan Kang3, Hak-Joon Sung3,4, Soon Hyun Ahn5, Eun Chang Choi2, Won Shik Kim5 1Department of Otolaryngology, Head and Neck Surgery, King Abdullah Medical City, 2Department of Otorhinolaryngology, Yonsei University, College of Medicine, 3Severance Biomedical Science Institute, Yonsei University, College of Medicine, 4The George W.Woodruff School of Mechanical Engineering, Georgia Institute of Technology, 5Department of Otorhinolaryngology-Head and Neck Surgery, Seoul National University College of Medicine Here we present a protocol for the use of the pig superior epigastric artery perforator flap as a learning module for head and neck microvascular reconstruction. Neuroscience Novel Passive Clearing Methods for the Rapid Production of Optical Transparency in Whole CNS Tissue Jiwon Woo1,2,3, Eunice Yoojin Lee4, Hyo-Suk Park1,3, Jeong Yoon Park1,3, Yong Eun Cho1,2,3 1Department of Neurosurgery, Gangnam Severance Hospital, Yonsei University College of Medicine, 2Brain Korea 21 PLUS Project for Medical Science, Yonsei University, 3The Spine and Spinal Cord Institute, Biomedical Center, Gangnam Severance Hospital, Yonsei University College of Medicine, 4Columbia University College of Physicians and Surgeons Here, we present two novel methodologies, psPACT and mPACT, for achieving maximal optical transparency and subsequent microscopic analysis of tissue vasculature in the intact rodent whole CNS. Immunology and Infection Two-photon Intravital Imaging of Leukocytes During the Immune Response in Lipopolysaccharide-treated Mouse Liver Sang A Park*1,2, Young Ho Choe*1,2, Sung Hwan Lee3, Young-Min Hyun1,2 1Department of Anatomy, Yonsei University College of Medicine, 2Brain Korea 21 Plus Project for Medical Science, Yonsei University College of Medicine, 3Department of Surgery, Yonsei University College of Medicine We established a novel surgical protocol for two-photon imaging of live mice liver with minimal invasion. With this technique, we identified the detailed structure of the liver during lipopolysaccharide-induced endotoxemia. We anticipate that this method may be utilized to determine the effectiveness of various reagents treatment to hepatic leukocyte migration. Neuroscience Neurobehavioral Assessments in a Mouse Model of Neonatal Hypoxic-ischemic Brain Injury MinGi Kim1,2, Ji Hea Yu1, Jung Hwa Seo1,2, Yoon-Kyum Shin1,2, Soohyun Wi1,2, Ahreum Baek1,3, Suk-Young Song1,5, Sung-Rae Cho1,2,4,5 1Department and Research Institute of Rehabilitation Medicine, Yonsei University College of Medicine, 2Brain Korea 21 PLUS Project for Medical Science, Yonsei University, 3Department of Rehabilitation Medicine, Yonsei University Wonju College of Medicine, 4Rehabilitation Institute of Neuromuscular Disease, Yonsei University College of Medicine, 5Graduate Program of NanoScience and Technology, Yonsei University We performed unilateral carotid artery occlusion on postnatal day 7-10 CD-1 mouse pups to create a neonatal hypoxic-ischemic (HI) model and investigated the effects of HI brain injury. We studied neurobehavioral functions in these mice compared to non-operated normal mice. Bioengineering A Protocol for Multiple Gene Knockout in Mouse Small Intestinal Organoids Using a CRISPR-concatemer Alessandra Merenda1,2, Amanda Andersson-Rolf1,2, Roxana C. Mustata1, Taibo Li1, Hyunki Kim3, Bon-Kyoung Koo1,2 1Wellcome Trust - Medical Research Council Stem Cell Institute, University of Cambridge, 2Department of Genetics, University of Cambridge, 3Department of Pathology, Yonsei University College of Medicine This protocol describes the steps for cloning multiple single guide RNAs into one guide RNA concatemer vector, which is of particular use in creating multi-gene knockouts using CRISPR/Cas9 technology. The generation of double knockouts in intestinal organoids is shown as a possible application of this method. Developmental Biology Visualization and Quantitative Analysis of Embryonic Angiogenesis in Xenopus tropicalis Jiyeon Ohk1, Hosung Jung1,2 1Brain Korea 21 PLUS Project for Medical Science, Yonsei University College of Medicine, 2Department of Anatomy, Brain Research Institute, Yonsei University College of Medicine This protocol demonstrates a fluorescence-based method to visualize the vasculature and to quantify its complexity in Xenopus tropicalis. Blood vessels can be imaged minutes after the injection of a fluorescent dye into the beating heart of an embryo after genetic and/or pharmacological manipulations to study cardiovascular development in vivo. Environment Enhancement of the Initial Growth Rate of Agricultural Plants by Using Static Magnetic Fields Seung C. Kim1, Alex Mason2, Wooseok Im3 1College of Medicine, Yonsei University, 2Simon Fraser University, 3Biomedical Research Institute, Seoul National University Hospital The goal of this protocol is to demonstrate the acceleration of the initial growth rate of plants by applying static magnetic fields with no external energy.