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National Yang-Ming University

10 ARTICLES PUBLISHED IN JoVE

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Developmental Biology

Organotypic Slice Cultures for Studies of Postnatal Neurogenesis
Adam J. Mosa 1, Sabrina Wang 2,3, Yao Fang Tan 1, J. Martin Wojtowicz 1
1Department of Physiology, University of Toronto, 2Institute of Anatomy and Cell Biology, School of Medicine, National Yang-Ming University, 3Department of Education and Research, Taipei City Hospital

Here we describe a technique for studying hippocampal postnatal neurogenesis using the organotypic slice culture technique. This method allows for in vitro manipulation of adult neurogenesis and allows for the direct application of pharmacological agents to the cultured hippocampus.

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Bioengineering

Electrotaxis Studies of Lung Cancer Cells using a Multichannel Dual-electric-field Microfluidic Chip
Hsien-San Hou 1, Hui-Fang Chang 1, Ji-Yen Cheng 1,2,3,4,5
1Research Center for Applied Sciences, Academia Sinica, 2Institute of Biophotonics, National Yang-Ming University, 3Biophotonics & Molecular Imaging Research Center (BMIRC), National Yang-Ming University, 4Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University, 5Ph.D. Program in Microbial Genomics, National Chung Hsing University

Many microfluidic devices have been developed for use in the study of electrotaxis. Yet, none of these chips allows the efficient study of the simultaneous chemical and electric-field (EF) effects on cells. We developed a polymethylmethacrylate-based device that offers better-controlled coexisting EF and chemical stimulation for use in electrotaxis research.

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Bioengineering

Designing Microfluidic Devices for Studying Cellular Responses Under Single or Coexisting Chemical/Electrical/Shear Stress Stimuli
Tzu-Yuan Chou 1, Yung-Shin Sun 2, Hsien-San Hou 3, Shang-Ying Wu 1, Yun Zhu 1, Ji-Yen Cheng 3, Kai-Yin Lo 1
1Department of Agricultural Chemistry, National Taiwan University, 2Department of Physics, Fu-Jen Catholic University, 3Research Center for Applied Sciences, Academia Sinica

Micro-fabricated devices integrated with fluidic components provide an in vitro platform for cell studies mimicking the in vivo micro-environment. We developed polymethylmethacrylate-based microfluidic chips for studying cellular responses under single or coexisting chemical/electrical/shear stress stimuli.

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Developmental Biology

Long-term Live Imaging of Drosophila Eye Disc
Chia-Kang Tsao 1,2, Hui-Yu Ku 1,2, Y. Henry Sun 1,2
1Institute of Genomic Sciences, National Yang-Ming University, 2Institute of Molecular Biology, Academia Sinica

This protocol describes a detailed method for the long-term ex vivo culture and live imaging of a Drosophila imaginal disc. It demonstrates photoreceptor differentiation and ommatidial rotation within the 10 h period of live imaging of the eye disc. The protocol is simple and does not require expensive setup.

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Biology

In Vitro SUMOylation Assay to Study SUMO E3 Ligase Activity
Wan-Shan Yang 1, Mel Campbell 2, Hsing-Jien Kung 2,3,4,5, Pei-Ching Chang 1,6
1Institute of Microbiology and Immunology, National Yang-Ming University, 2UC Davis Cancer Center, University of California, Davis, 3Department of Biochemistry and Molecular Medicine, University of California, Davis, 4Institute for Translational Medicine, College of Medical Science and Technology, Taipei Medical University, 5Division of Molecular and Genomic Medicine, National Health Research Institutes, 6Center for Infectious Disease and Cancer Research, Kaohsiung Medical University

Unlike ubiquitin ligases, few E3 SUMO ligases have been identified. This modified in vitro SUMOylation protocol is able to identify novel SUMO E3 ligases by an in vitro reconstitution assay.

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Neuroscience

Stereotaxic Surgery for Genetic Manipulation in Striatal Cells of Neonatal Mouse Brains
Shih-Yun Chen 1, Hsiao-Ying Kuo 1, Fu-Chin Liu 1,2
1Institute of Neuroscience, National Yang-Ming University, 2Brain Research Center, National Yang-Ming University

We describe a protocol of stereotaxic surgery with a homemade head-fixed device for microinjecting reagents into the striatum of neonatal mouse brains. This technique allows genetic manipulation in neuronal cells of specific regions of neonatal mouse brains.

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Immunology and Infection

Multicolor Flow Cytometry-based Quantification of Mitochondria and Lysosomes in T Cells
Chin-Wen Wei 1, Tyng-An Zhou 1, Ivan L. Dzhagalov 1, Chia-Lin Hsu 1
1Institute of Microbiology and Immunology, National Yang-Ming University

This article illustrates a powerful method to quantify mitochondria or lysosomes in living cells. The combination of lysosome- or mitochondria-specific dyes with fluorescently conjugated antibodies against surface markers allows the quantification of these organelles in mixed cell populations, like primary cells harvested from tissue samples, by using multicolor flow cytometry.

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Bioengineering

Wet-spinning-based Molding Process of Gelatin for Tissue Regeneration
Chia-Yu Wang 1, Dewi Sartika 2, Ding-Han Wang 3, Po-Da Hong 1, Juin-Hong Cherng 2,4,5, Shu-Jen Chang 2,6, Cheng-Che Liu 7, Yi-Wen Wang 4, Sheng-Tang Wu 8
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.

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Medicine

Sleeve Gastrectomy in Mice using Surgical Clips
Jih-Hua Wei 1,2,3, Che-Hung Yeh 4, Wei-Jei Lee 5, Shing-Jong Lin 3,6,7,8,9, Po-Hsun Huang 3,6,7
1Cardiovascular Division, Internal Medicine Department, Min-Sheng General Hospital, 2Department of Nutrition and Health Sciences, School of Healthcare Management, Kai-Nan University, 3Institute of Clinical Medicine, National Yang-Ming University, 4Food Science and Biotechnology, College of Biotechnology and Bioresources, University of Da-Yeh, 5Department of Surgery, Min-Sheng General Hospital, 6Department of Critical Care Medicine, Taipei Veterans General Hospital, 7Division of Cardiology, Department of Internal Medicine, Taipei Veterans General Hospital, 8Division of Cardiology, Heart Center, Cheng-Hsin General Hospital, 9Taipei Medical University

The prevalence of diabetes and obesity is continuously increasing worldwide. The mechanisms between diabetes, obesity and their associated mortality and co-morbidities need to be further investigated. Here, we present a protocol for sleeve gastrectomy (SG) in animals as an uncomplicated preclinical model of bariatric surgery.

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Bioengineering

Electric-Field-Induced Neural Precursor Cell Differentiation in Microfluidic Devices
Hui-Fang Chang 1, Shih-En Chou 1, Ji-Yen Cheng 1,2,3,4
1Research Center for Applied Sciences, Academia Sinica, 2Institute of Biophotonics, National Yang Ming Chao Tung University, 3Department of Mechanical and Mechatronic Engineering, National Taiwan Ocean University, 4College of Engineering, Chang Gung University

In this study, we present a protocol for the differentiation of neural stem and progenitor cells (NPCs) solely induced by direct current (DC) pulse stimulation in a microfluidic system.

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