National Defense Medical Center 6 articles published in JoVE Neuroscience Two-Photon Calcium Imaging of Forebrain Activity in Behaving Adult Zebrafish Jose Sandino Bandonil*1,2, Yi-Hsuan Liao*1, Amir Fathi1, Kuo-Hua Huang1,2 1Institute of Molecular Biology, Academia Sinica, 2Molecular and Cell Biology, Taiwan International Graduate Program, Academia Sinica and Graduate Institute of Life Sciences, National Defense Medical Center Here, we present a protocol to perform two-photon calcium imaging in the dorsal forebrain of adult zebrafish. Bioengineering Wet-spinning-based Molding Process of Gelatin for Tissue Regeneration Chia-Yu Wang1, Dewi Sartika2, Ding-Han Wang3, Po-Da Hong1, Juin-Hong Cherng2,4,5, Shu-Jen Chang2,6, Cheng-Che Liu7, Yi-Wen Wang4, Sheng-Tang Wu8 1Department of Materials Science and Engineering, National Taiwan University of Science and Technology, 2Laboratory of Adult Stem Cell and Tissue Regeneration, National Defense Medical Center, 3School of Dentistry, National Yang-Ming University, 4Department and Graduate Institute of Biology and Anatomy, National Defense Medical Center, 5Department of Gerontological Health Care, National Taipei University of Nursing and Health Sciences, 6Division of Rheumatology/Immunology/Allergy, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, 7Department of Physiology and Biophysics, Graduate Institute of Physiology, National Defense Medical Center, 8Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center We developed and describe a protocol based on the wet spinning concept, for the construction of gelatin-based biomaterials used for the application of tissue engineering. Medicine Severe Burn Injury in a Swine Model for Clinical Dressing Assessment Gang-Yi Fan1, Juin-Hong Cherng2,3, Shu-Jen Chang4, Raju Poongodi2, Chih-Hsin Wang5, Yi-Wen Wang2, Yao-Horng Wang6, Cheng-Che Liu7, En Meng8 1Graduate Institute of Medical Sciences, National Defense Medical Center, 2Department and Graduate Institute of Biology and Anatomy, National Defense Medical Center, 3Department of Gerontological Health Care, National Taipei University of Nursing and Health Sciences, 4Division of Rheumatology/Immunology/Allergy, Department of Internal Medicine, Tri-Service General Hospital, National Defense Medical Center, 5Department of Plastic and Reconstructive Surgery, Tri-Service General Hospital, 6Department of Nursing, Yuanpei University, 7Department of Physiology and Biophysics; Graduate Institute of Physiology, National Defense Medical Center, 8Division of Urology, Department of Surgery, Tri-Service General Hospital, National Defense Medical Center To closely mimic the mode of burn injuries requires the interplay between clinical observation and studies in animal models. In this study, a swine model of severe burn injury was established to assess an experimental dressing in physiological and pathophysiological settings. Neuroscience Visualization of Motor Axon Navigation and Quantification of Axon Arborization In Mouse Embryos Using Light Sheet Fluorescence Microscopy Ee Shan Liau*1,2, Ya-Ping Yen*2,3, Jun-An Chen1,2 1Molecular and Cell Biology, Taiwan International Graduate Program, Academia Sinica and Graduate Institute of Life Sciences, National Defense Medical Center, 2Institute of Molecular Biology, Academia Sinica, 3Institute of Biotechnology, College of Bio-Resources and Agriculture, National Taiwan University Here, we describe a protocol for visualizing motor neuron projection and axon arborization in transgenic Hb9::GFP mouse embryos. After immunostaining for motor neurons, we used light sheet fluorescence microscopy to image embryos for subsequent quantitative analysis. This protocol is applicable to other neuron navigation processes in the central nervous system. Developmental Biology Cell Lineage Analyses and Gene Function Studies Using Twin-spot MARCM Hung-Chang Shen1,2, Tsai-Chi Hsu1, Pei-Chi Chung1, Hung-Hsiang Yu1,2 1Institute of Cellular and Organismic Biology, Academia Sinica, 2Graduate Institute of Life Sciences, National Defense Medical Center Here, we present a protocol for a mosaic labeling technique that permits the visualization of neurons derived from a common progenitor cell in two distinct colors. This facilitates neural lineage analysis with the capability of birth-dating individual neurons and studying gene function in the same neurons of different individuals. Immunology and Infection Assessment of the Immunomodulatory Properties of Human Mesenchymal Stem Cells (MSCs) Pei-Ju Hsu1, Ko-Jiunn Liu2, Ying-Yin Chao1, Huey-Kang Sytwu3, B. Linju Yen1 1Regenerative Medicine Research Group, Institute of Cellular & System Medicine, National Health Research Institutes (NHRI), 2National Institute of Cancer Research, National Health Research Institutes (NHRI), 3Institute of Microbiology & Immunology, National Defense Medical Center The immunomodulatory properties of human mesenchymal stem cells (MSC) appear increasingly relevant for clinical application. Using a co-culture system of MSCs and peripheral blood leukocytes pre-stained with the fluorescent dye carboxyfluorescein succinimidyl ester (CFSE), we describe the in vitro assessment of MSC immunomodulation on effector leukocyte proliferation and specific subpopulations.