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National Health Research Institutes

7 ARTICLES PUBLISHED IN JoVE

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JoVE Journal

Quantitative Analysis of Autophagy using Advanced 3D Fluorescence Microscopy
Chun A. Changou 1,2, Deanna L. Wolfson 2, Balpreet Singh Ahluwalia 2,3, Richard J. Bold 4,5, Hsing-Jien Kung 5,6, Frank Y.S. Chuang 1,2,5
1Department of Biochemistry and Molecular Medicine, University of California, Davis , 2NSF Center for Biophotonics Science & Technology, University of California, Davis , 3University of Tromsø, 4Department of Surgery (Division of Surgical Oncology), University of California, Davis , 5UC Davis Comprehensive Cancer Center, University of California, Davis , 6Department of Biological Chemistry, University of California, Davis

Autophagy is a ubiquitous process that enables cells to degrade and recycle proteins and organelles. We apply advanced fluorescence microscopy to visualize and quantify the small, but essential, physical changes associated with the induction of autophagy, including the formation and distribution of autophagosomes and lysosomes, and their fusion into autolysosomes.

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Bioengineering

A Microfluidic Platform for High-throughput Single-cell Isolation and Culture
Ching-Hui Lin 1,2, Hao-Chen Chang 1,2, Chia-Hsien Hsu 1,2,3
1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, Taiwan, 2Tissue Engineering and Regenerative Medicine, National Chung Hsing University, 3Institute of NanoEngineering and MicroSystems, National Tsing Hua University

Here, we present a protocol for isolating and culturing single cells with a microfluidic platform, which utilizes a new microwell design concept to allow for high-efficiency single cell isolation and long-term clonal culture.

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Bioengineering

An Ultrasonic Tool for Nerve Conduction Block in Diabetic Rat Models
Yee Fun Lee 1, Chou-Ching Lin 2, Jung-Sung Cheng 1, Gin-Shin Chen 1
1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, 2Department of Neurology, National Cheng Kung University Hospital

This work presents the methodology of applying high intensity-focused ultrasound to block the action potentials of diabetic neuropathic nerves.

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Developmental Biology

Transfer of Mammary Gland-forming Ability Between Mammary Basal Epithelial Cells and Mammary Luminal Cells via Extracellular Vesicles/Exosomes
Meng-Chieh Lin *1, Shih-Yin Chen *1, Pei-Lin He 1, Wen-Ting Luo 1, Hua-Jung Li 1
1Institute of Cellular and System Medicine, National Health Research Institutes

This protocol describes methods for purifying, quantitating, and characterizing extracellular vesicles (EVs)/exosomes from non-adherent/mesenchymal mammary epithelial cells and for using them to transfer mammary gland-forming ability to luminal mammary epithelial cells. EVs/exosomes derived from stem-like mammary epithelial cells can transfer this cell property to cells that ingest the EVs/exosomes.

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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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Bioengineering

Establishing Single-Cell Based Co-Cultures in a Deterministic Manner with a Microfluidic Chip
Cheng-Kun He 1,2, Ya-Wen Chen 3, Ssu-Han Wang 3, Chia-Hsien Hsu 1,2
1Institute of Biomedical Engineering and Nanomedicine, National Health Research Institutes, 2Ph.D. Program in Tissue Engineering and Regenerative Medicine, National Chung Hsing University, 3National Institute of Cancer Research, National Health Research Institutes

This report describes a microfluidic chip-based method to set up a single cell culture experiment in which high-efficiency pairing and microscopic analysis of multiple single cells can be achieved.

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Biology

Induction of Petite Colonies in Candida glabrate via Rose Bengal-Mediated Photodynamic Therapy
Cai-Ying Yang *1,2, Jia-Horung Hung *3, Chi-Jung Wu 4,5, Zhao-Xiang Wang 1,2, Shih-Han Wang 6, Hao-Chun Liaw 2, I-Huang Lin 3, Chun-Keung Yu 1,6, Tak-Wah Wong 2,7,8
1Department of Microbiology and Immunology, College of Medicine, National Cheng Kung University, 2Department of Dermatology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, 3Department of Ophthalmology, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, 4National Institute of Infectious Diseases and Vaccinology, National Health Research Institutes, 5Department of Internal Medicine, National Cheng Kung University Hospital, College of Medicine, National Cheng Kung University, 6Insititute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, 7Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, 8Center of Applied Nanomedicine, National Cheng Kung University

The significance of petite colonies in Candida spp. drug resistance has not been fully explored. Antimicrobial photodynamic therapy (aPDT) offers a promising strategy against drug-resistant fungal infections. This study demonstrates that rose bengal-mediated aPDT effectively deactivates Candida glabrata and induces petite colonies, presenting a unique procedure.

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