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51 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Analyzing Mixing Inhomogeneity in a Microfluidic Device by Microscale Schlieren Technique
Chen-li Sun 1, Tzu-hsun Hsiao 2
1Department of Mechanical Engineering, National Taiwan University, 2Department of Mechanical Engineering, National Taiwan University of Science and Technology

Herein, we describe a procedure that employs microscale schlieren technique to measure mixing inhomogeneity in a microfluidic device. Through calibration, distribution of concentration gradient can be derived from the micro-schlieren image.

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Engineering

Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle
Hsuan Lee 1, Kuan-Yu Li 1, Yen-Ta Huang 1, Po-Ting Shen 1, Gitanjal Deka 1, Ryosuke Oketani 2, Yasuo Yonemaru 2, Masahito Yamanaka 2, Katsumasa Fujita 2, Shi-Wei Chu 1,3
1Department of Physics, National Taiwan University, 2Department of Applied Physics, Osaka University, 3Molecular Imaging Center, National Taiwan University

Saturable and reverse saturable scattering were discovered in isolated plasmonic particles and adopted as a novel non-bleaching contrast method in super-resolution microscopy. Here the experimental procedures of detecting and extracting nonlinear scattering are explained in detail, as well as how to enhance resolution with the aid of saturated excitation microscopy.

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Biology

Development of a Colloidal Gold-based Immunochromatographic Test Strip for Detection of Cetacean Myoglobin
Kun-Wei Chan 1, Chieh Lo 1, Chi-Shih Chu 2, Li-Te Chin 2, Yu-Ting Wang 1, Wei-Cheng Yang 1
1Department of Veterinary Medicine, National Chiayi University, 2Department of Microbiology and Immunology/Graduate Institute of Biomedical and Biopharmaceutical Sciences, National Chiayi University

This protocol describes the development of two IgG class monoclonal antibodies (mAbs) strongly reactive to myoglobin of cetaceans. These mAbs are applied on a colloidal gold immunochromatographic test strip based on the sandwich format to differentiate the Mb of cetaceans from seal and other animals.

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Education

A Fabrication Method for Highly Stretchable Conductors with Silver Nanowires
Chia-Wei Chang 1, Shih-Pin Chen 1, Ying-Chih Liao 1
1Department of Chemical Engineering, National Taiwan University

A simple synthesis method is used to chemically solder silver nanowire thin film to fabricate highly stretchable and conductive metal conductors.

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Developmental Biology

Primary Endodermal Epithelial Cell Culture from the Yolk Sac Membrane of Japanese Quail Embryos
Han Jen Lin 1, Siou Huei Wang 1, Yu Hui Pan 1, Shih-Torng Ding 1
1Department of Animal Science and Technology, National Taiwan University

To study the mechanism of lipid utilization in yolk sac membranes during the late stages of avian embryonic development, we established a primary Japanese quail embryonic endodermal epithelial cell culture system.

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Behavior

Comparing the Frequency Effect Between the Lexical Decision and Naming Tasks in Chinese
Xin-Yu Gao *1, Meng-Feng Li *1, Tai-Li Chou *2, Jei-Tun Wu 1
1Department of Psychology, National Taiwan University, 2Neurobiology and Cognitive Science Center, National Taiwan University

Researchers adopt both the lexical decision task and the naming task to investigate some important topics such as character/word recognition by comparing the frequency effect between these two tasks. This article introduces this approach through two exemplar experiments and elaborates on the underlying logic.

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Developmental Biology

Identification of Critical Conditions for Immunostaining in the Pea Aphid Embryos: Increasing Tissue Permeability and Decreasing Background Staining
Gee-Way Lin 1,2, Chun-che Chang 1,2,3,4
1Department of Entomology, National Taiwan University, 2Institute of Biotechnology, National Taiwan University, 3Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, 4Genome and Systems Biology Degree Program, National Taiwan University and Academia Sinica

A protocol of whole-mount immunostaining on aphid embryos is presented, in which critical conditions for decreasing background staining and increasing tissue permeability are addressed. These conditions are specifically developed for effective detection of protein expression in the embryonic tissues of aphids.

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Developmental Biology

Isolation and Differentiation of Adipose-Derived Stem Cells from Porcine Subcutaneous Adipose Tissues
Yu-Jen Chen 1,2, Hui-Yu Liu 2, Yun-Tsui Chang 2, Ying-Hung Cheng 2, Harry J. Mersmann 2, Wen-Hung Kuo 3, Shih-Torng Ding 1,2
1Institute of Biotechnology, National Taiwan University, 2Department of Animal Science and Technology, National Taiwan University, 3Department of Surgery, National Taiwan University Hospital and College of Medicine

This protocol describes the isolation of pig adipose-derived stem cells (pADSC) from subcutaneous adipose tissues with examination of multipotency. The multipotent pADSC are used to delineate processes of adipocyte differentiation and study transdifferentiation into multiple cell lineages of mesodermal mesenchyme or further lineages of ectoderm and endoderm for regenerative studies.

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Immunology and Infection

Intravital Imaging of Neutrophil Priming Using IL-1β Promoter-driven DsRed Reporter Mice
Yi Yao 1,2, Yun Liu 1,3, Akira Takashima 1
1Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, 2Department of Internal Medicine, Yale University School of Medicine, 3Department of Pathophysiology, Southern Medical University (China)

This current protocol employs fluorescent reporters, in vivo labeling, and intravital imaging techniques to enable monitoring of the dynamic process of neutrophil priming in living animals.

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Environment

Clean Sampling and Analysis of River and Estuarine Waters for Trace Metal Studies
Kuo-Tung Jiann *1, Liang-Saw Wen *2,3, Peter H. Santschi 4
1Department of Oceanography, National Sun Yat-sen University, 2Institute of Oceanography, National Taiwan University, 3Taiwan Ocean Research Institute, National Research Laboratories, 4Department of Oceanography and Marine Sciences, Texas A&M University at Galveston

Special care using "clean techniques" is required to properly collect and process water samples for trace metal studies in aquatic environments. A protocol for sampling, processing, and analytical procedures with the aim of obtaining reliable environmental monitoring data and results with high sensitivity for detailed trace metal studies is presented.

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Developmental Biology

In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition
Wei-Ting Ho 1,2, Chien-Chia Su 3, Jung-Shen Chang 3, Shu-Wen Chang 1, Fung-Rong Hu 3, Tzuu-Shuh Jou 2,4, I-Jong Wang 3,4
1Department of Ophthalmology, Far Eastern Memorial Hospital, 2Institute of Clinical Medicine, National Taiwan University, 3Department of Ophthalmology, National Taiwan University Hospital, 4College of Medicine, National Taiwan University

A primary culture of bovine corneal endothelial cells was used to investigate the mechanism of corneal endothelial-mesenchymal transition. Furthermore, a rat corneal endothelium cryoinjury model was used to demonstrate corneal endothelial-mesenchymal transition in vivo.

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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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Biochemistry

Preparation of Homogeneous MALDI Samples for Quantitative Applications
Yu-Meng Ou 1,2, Chien-Wei Tsao 1,3, Yin-Hung Lai 1, Hsun Lee 1, Huan-Tsung Chang 2, Yi-Sheng Wang 1
1Genomic Research Center, Academia Sinica, 2Department of Chemistry, National Taiwan University, 3Department of Chemical Engineering and Biotechnology, National Taipei University of Technology

A protocol for reducing spatial heterogeneities of ion signals in MALDI mass spectrometry by regulating substrate temperature during sample drying processes is demonstrated.

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Genetics

The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform
Szu-I Yeh 1, Wei-Feng Fang 1, Chao-Jyun Huang 1, Tzu-Ming Wang 1, Jing-Tang Yang 1
1Department of Mechanical Engineering, National Taiwan University

This work presents a simple and visual method to detect multi-nucleotide polymorphisms on a pneumatic droplet manipulation platform. With the proposed method, the entire experiment, including droplet manipulation and detection of multi-nucleotide polymorphisms, can be performed near 23 °C without the aid of advanced instruments.

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Immunology and Infection

The Isolation, Differentiation, and Quantification of Human Antibody-secreting B Cells from Blood: ELISpot as a Functional Readout of Humoral Immunity
Shiang-Jong Tzeng 1
1Graduate Institute of Pharmacology, College of Medicine, National Taiwan University

Human peripheral blood is commonly used for the assessment of the humoral immune response. Here, the methods for isolating human B cells from peripheral blood, differentiating human B cells into antibody (Ab)-secreting B cells (ASCs) in culture, and enumerating the total IgM- and IgG-ASCs via an ELISpot assay are described.

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Medicine

A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats
Hui-Chun Ku 1, Shih-Yi Lee 2,3, Yuan-Kun Aden Wu 4, Kai-Chien Yang 1,5, Ming-Jai Su 1
1Graduate Institute of Pharmacology, College of Medicine, National Taiwan University, 2Division of Pulmonary and Critical Care Medicine, Mackay Memorial Hospital, 3Mackay Medicine, Nursing and Management College, 4School of Medicine, National Taiwan University, 5Department of Internal Medicine, National Taiwan University Hosptial

A rat model of abdominal aortic constriction that induces cardiac hypertrophy and remodeling is described. An efficient, highly-reproducible, and minimally-invasive method is used to provide a simple yet useful platform for research in myocardial hypertrophy and dysfunction.

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Behavior

Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture
Pei-Lu Yi *1, Shuo-Bin Jou *2, Yi-Jou Wu 3, Fang-Chia Chang 3,4,5
1Department of Sports, Health & Leisure, College of Tourism, Leisure and Sports, Aletheia University, Tainan Campus, 2Department of Neurology, Mackay Memorial Hospital and Mackay Medical College, 3Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University, 4Graduate Institute of Brain & Mind Sciences, College of Medicine, National Taiwan University, 5Graduate Institute of Acupuncture Science, College of Chinese Medicine, China Medical University

This paper demonstrates the performance of acupuncture, epilepsy models, and the analysis of sleep in rodents. The acupuncture procedure and the identification of acupoints are described. Pilocarpine or pentylenetetrazol (PTZ) is used to induce epilepsy. Electrocorticogram (ECoG), electromyogram (EMG), brain temperature, and locomotor activity recordings are employed for sleep analysis.

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Engineering

Pool-Boiling Heat-Transfer Enhancement on Cylindrical Surfaces with Hybrid Wettable Patterns
Sujith Kumar C.S. *1, Yao Wen Chang *1, Ping-Hei Chen 1
1Department of Mechanical Engineering, National Taiwan University

Pool-boiling heat-transfer experiments were carried out to observe the effects of hybrid wettable patterns on the heat-transfer coefficient (HTC). The parameters of investigation are the number of interlines and the pattern orientation of the modified wettable surface.

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Bioengineering

Magnetic and Thermal-sensitive Poly(N-isopropylacrylamide)-based Microgels for Magnetically Triggered Controlled Release
Chih-Yu Kuo 1, Ting-Yu Liu 2, Kuan-Syun Wang 2, Andri Hardiansyah 3, Yen-Ting Lin 4, Hsueh-Yung Chen 4, Wen-Yen Chiu 1,4,5
1Institute of Polymer Science and Engineering, National Taiwan University, 2Department of Materials Engineering, Ming Chi University of Technology, 3Department of Metallurgy and Materials Engineering, Bandung Institute of Technology and Science, 4Department of Chemical Engineering, National Taiwan University, 5Department of Materials Science and Engineering, National Taiwan University

This manuscript describes the preparation of magnetic and thermal-sensitive microgels via a temperature-induced emulsion without chemical reaction. These sensitive microgels were synthesized by mixing poly(N-isopropylacrylamide) (PNIPAAm), polyethylenimine (PEI) and Fe3O4-NH2 nanoparticles for the potential use in magnetically and thermally triggered drug-release.

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Bioengineering

An Integrated System to Remotely Trigger Intracellular Signal Transduction by Upconversion Nanoparticle-mediated Kinase Photoactivation
Hua-De Gao 1,2, Pounraj Thanasekaran 1, Tzu-Ho Chen 1,2, Yu-Hsu Chang 3, Yu-Ju Chen 1,2, Hsien-Ming Lee 1
1Institute of Chemistry, Academia Sinica, 2Department of Chemistry, National Taiwan University, 3Department of Materials and Mineral Resources Engineering, National Taipei University of Technology

In this protocol, caged protein kinase A (PKA), a cellular signal transduction bioeffector, was immobilized on a nanoparticle surface, microinjected into the cytosol, and activated by the upconverted UV light from near-infrared (NIR) irradiation, inducing downstream stress fiber disintegration in the cytosol.

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JoVE Core

Preparation of Janus Particles and Alternating Current Electrokinetic Measurements with a Rapidly Fabricated Indium Tin Oxide Electrode Array
Yu-Liang Chen 1, Hong-Ren Jiang 1
1Institute of Applied Mechanics, National Taiwan University

In this article, a simple method to prepare partially or fully coated metallic particles and to perform AC electrokinetic property measurements with a rapidly fabricated indium tin oxide (ITO) electrode array is demonstrated.

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Engineering

Well-aligned Vertically Oriented ZnO Nanorod Arrays and their Application in Inverted Small Molecule Solar Cells
Ming-Yi Lin 1, Shang-Hsuan Wu 2, Li-Jen Hsiao 3, Widhya Budiawan 2, Shih-Lun Chen 4, Wei-Chen Tu 4, Chia-Yen Lee 1, Yia-Chung Chang 2, Chih-Wei Chu 2
1Department of Electrical Engineering, National United University, 2Research Center of Applied Science, Academia Sinica, 3Department of Graduate Institute of Photonics and Optoelectronics, National Taiwan University, 4Department of Electronic Engineering, Chung Yuan Christian University

This manuscript describes how to design and fabricate efficient inverted SMPV1:PC71BM solar cells with ZnO nanorods (NRs) grown on a high quality Al-doped ZnO (AZO) seed layer. The well-aligned vertically oriented ZnO NRs exhibit high crystalline properties. The power conversion efficiency of solar cells can reach 6.01%.

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Engineering

Preparation of Large-area Vertical 2D Crystal Hetero-structures Through the Sulfurization of Transition Metal Films for Device Fabrication
Chong-Rong Wu 1,2, Tung-Wei Chu 2,3, Kuan-Chao Chen 1,2, Shih-Yen Lin 1,2
1Graduate Institute of Electronics Engineering, National Taiwan University, 2Research Center for Applied Sciences, Academia Sinica, 3Graduate Institute of Photonics and Optoelectronics, National Taiwan University

Through the sulfurization of pre-deposited transition metals, large-area and vertical 2D crystal hetero-structures can be fabricated. The film transferring and device fabrication procedures are also demonstrated in this report.

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Biology

3D Mitochondrial Ultrastructure of Drosophila Indirect Flight Muscle Revealed by Serial-section Electron Tomography
Yi-fan Jiang 1, Hsiang-ling Lin 2, Chi-yu Fu 2
1Graduate Institute of Molecular and Comparative Pathobiology, School of Veterinary Medicine, National Taiwan University, 2Institute of Cellular and Organismic Biology, Academia Sinica

In this protocol, we demonstrate the application of serial-section electron tomography to elucidate mitochondrial structure in Drosophila indirect flight muscle.

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Genetics

Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay
Ju-Chun Chang 1, Se Jin Lee 2, Jae Su Kim 2, Chung-Hsiung Wang 3, Yu-Shin Nai 1
1Department of Biotechnology and Animal Science, National Ilan University, 2Department of Agricultural Biology, College of Agriculture Life Sciences, Chonbuk National University, 3Department of Entomology, National Taiwan University

This protocol describes a heat shock-induced protein expression system (pDHsp/V5-His/sf9 cell system), which can be used for either expressing foreign proteins or evaluating the anti-apoptotic activity of potential foreign proteins and their truncated amino acids in insect cells.

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Cancer Research

The Establishment of a Lung Colonization Assay for Circulating Tumor Cell Visualization in Lung Tissues
Tsung-Cheng Lin 1, Ying-Chih Liao 2, Wen-Tsan Chang 1,2, Cheng-Han Yang 1, Li-Hsin Cheng 1, Megan Cheng 3, Hung-Chi Cheng 1,2
1The Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, 2Department of Biochemistry and Molecular Biology, College of Medicine, National Cheng Kung University, 3Trauma Office, Children's National Health System

An animal model is needed to decipher the role of circulating tumor cells (CTCs) in promoting lung colonization during cancer metastasis. Here, we established and successfully performed an in vivo assay to specifically test the requirement of polymeric fibronectin (polyFN) assembly on CTCs for lung colonization.

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Environment

An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles
Liang-Saw Wen 1, Chih-Ping Lee 1, Wen-Huei Lee 1, Alan Chuang 2
1Institute of Oceanography, National Taiwan University, 2Sino Instruments Co., Ltd.

Plankton and suspended particles play a major role in the biogeochemical cycles in the ocean. Here, we provide an ultra-clean, low stress method for the collection of various sizes of particles and plankton at sea with the capability of handling large volumes of seawater.

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Biology

Practical Use of RNA Interference: Oral Delivery of Double-stranded RNA in Liposome Carriers for Cockroaches
Jia-Hsin Huang 1, Yun Liu 1, Yu-Hsien Lin 1,2,3, Xavier Belles 4, How-Jing Lee 1
1Department of Entomology, National Taiwan University, 2Biology Centre, Institute of Entomology, Czech Academy of Sciences, 3Faculty of Science, University of South Bohemia, 4Institute of Evolutionary Biology, CSIC-UPF

This manuscript demonstrates the depletion of gene expression in the midgut of the German cockroach through oral ingestion of double-stranded RNA encapsulated in liposomes.

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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 Chen 1,2
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.

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Environment

A Novel Method for the Pentosan Analysis Present in Jute Biomass and Its Conversion into Sugar Monomers Using Acidic Ionic Liquid
Babasaheb M. Matsagar 1, Shahriar A. Hossain 2,3, Tofazzal Islam 4, Yusuke Yamauchi 2,3,5,6, Kevin C.-W. Wu 1
1Department of Chemical Engineering, National Taiwan University, 2International Center for Materials Nanoarchitectonics (MANA), National Institute for Materials Science (NIMS), 3Australian Institute for Innovative Materials (AIIM), University of Wollongong, 4Department of Biotechnology, Bangabandhu Sheikh Mujibur Rahman Agricultural University, 5School of Chemical Engineering & Australian Institute for Bioengineering and Nanotechnology (AIBN), University of Queensland, 6Department of Plant and Environmental New Resources, Kyung Hee University

We present a protocol for the synthesis of C5 sugars (xylose and arabinose) from a renewable non-edible lignocellulosic biomass (i.e., jute) with the presence of Brønsted acidic ionic liquids (BAILs) as the catalyst in water. The BAILs catalyst exhibited better catalytic performance than conventional mineral acid catalysts (H2SO4 and HCl).

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Chemistry

An Efficient Sample Preparation Method to Enhance Carbohydrate Ion Signals in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry
Yu-Meng Ou 1,2, Shu-Yun Kuo 1, Hsun Lee 1, Huan-Tsung Chang 2, Yi-Sheng Wang 1
1Genomic Research Center, Academia Sinica, 2Department of Chemistry, National Taiwan University

A protocol for enhancing carbohydrate ion signals in MALDI mass spectrometry by reforming crystalline structures during sample preparation processes is demonstrated.

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Education

Proofreading and DNA Repair Assay Using Single Nucleotide Extension and MALDI-TOF Mass Spectrometry Analysis
Kang-Yi Su *1,2, Steven D. Goodman *3, Hung-Ming Lai 1, Rong-Syuan Yen 1, Wei-Yao Hu 1, Wern-Cherng Cheng 2, Liang-In Lin 1,2, Ya-Chien Yang 1,2, Woei-Horng Fang 1,2
1Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, 2Department of Laboratory Medicine, National Taiwan University Hospital, 3Center for Microbial Pathogenesis, Nationwide Children's Hospital and the Department of Pediatrics, The Ohio State University

A non-labeled, non-radio-isotopic method for DNA polymerase proofreading and a DNA repair assay was developed by using high-resolution MALDI-TOF mass spectrometry and a single nucleotide extension strategy. The assay proved to be very specific, simple, rapid, and easy to perform for proofreading and repair patches shorter than 9-nucleotides.

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Behavior

Computerized Adaptive Testing System of Functional Assessment of Stroke
Gong-Hong Lin 1, Yi-Jing Huang 1, Yeh-Tai Chou 2, Hsin-Yu Chiang 3, Ching-Lin Hsieh 1,4,5
1School of Occupational Therapy, College of Medicine, National Taiwan University, 2Research Center for Psychological and Educational Testing, National Taiwan Normal University, 3Department of Occupational Therapy, College of Medicine, Fu Jen Catholic University, 4Department of Physical Medicine and Rehabilitation, National Taiwan University Hospital, 5Department of Occupational Therapy, College of Medical and Health Science, Asia University

Here, we present a protocol to develop the computerized adaptive testing system of the functional assessment of stroke (CAT-FAS). The CAT-FAS can simultaneously assess four functions (two motor functions [upper and lower extremities], postural control, and basic activities of daily living) with sufficient reliability and administrative efficiency.

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Environment

Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues
Wen-Ta Li 1, Bang-Yeh Liou 1, Wei-Cheng Yang 2, Meng-Hsien Chen 3, Hui-Wen Chang 1,2, Hue-Ying Chiou 4, Victor Fei Pang 1,2, Chian-Ren Jeng 1,2
1Graduate Institute of Molecular and Comparative Pathobiology, National Taiwan University, 2School of Veterinary Medicine, National Taiwan University, 3Department of Oceanography and Asia-Pacific Ocean Research Center, National Sun Yat-sen University, 4Graduate Institute of Veterinary Pathobiology, National Chung Hsing University

A protocol is presented to localize Ag in cetacean liver and kidney tissues by autometallography. Furthermore, a new assay, named the cetacean histological Ag assay (CHAA) is developed to estimate the Ag concentrations in those tissues.

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Chemistry

Extending the Lifespan of Soluble Lead Flow Batteries with a Sodium Acetate Additive
Yan-Ting Lin 1, Wei-Cheng Kuo 1, Chun-Yen Lee 1, Hao-Lun Tan 1, Hsun-Yi Chen 1, Ho-Wei Chan 1, Yong-Hong Lai 1, Kai-Rui Pan 1
1Bio-industrial Mechatronics Engineering Department, National Taiwan University

A protocol for the construction of a soluble lead flow battery with an extended lifespan, in which sodium acetate is supplied in the methanesulfonic electrolyte as an additive, is presented.

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Environment

Standard Operating Procedure for Lyssavirus Surveillance of the Bat Population in Taiwan
Wei-Cheng Hsu 1, Chao-Lung Hsu 2,3, Yang-Chang Tu 1, Jen-Chieh Chang 1, Kwok-Rong Tsai 1, Fan Lee 1, Shu-Chia Hu 1
1Animal Health Research Institute, Council of Agriculture, Executive Yuan, 2Institute of Ecology and Evolutionary Biology, National Taiwan University, 3Bat Conservation Society of Taipei

This protocol introduces a standard laboratory operating procedure for diagnostic testing of lyssavirus antigens in bats in Taiwan.

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Behavior

Examining Online Syntactic Processing of Spoken Complex Sentences in Chinese Using Dual-Modal Interference Tasks
Tuyuan Cheng 1, Jei-Tun Wu 2
1General Education Center, National Tainan Junior College of Nursing, 2Department of Psychology, National Taiwan University

Here, we present a protocol employing dual-modal interference tasks to examine online processing of spoken-Chinese relative clause sentences. Two exemplar experiments involving auditory processing with intra- and extrasentential interferences are described. The paradigm provides a methodology for addressing the nature of working memory and its effects on sentence processing.

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Medicine

Integration of Brain Tissue Saturation Monitoring in Cardiopulmonary Exercise Testing in Patients with Heart Failure
Shu-Chun Huang 1,2,3, Carl PC Chen 1,3, Tieh-Cheng Fu 4, Yu-Jen Chen 1,5,6
1Department of Physical Medicine and Rehabilitation, Chang Gung Memorial Hospital, Linkou, 2Healthy Aging Research Center, Chang Gung University, 3School of Medicine, College of Medicine, Chang Gung University, 4Department of Physical Medicine and Rehabilitation, Chang Gung Memorial Hospital, Keelung, 5National Quemoy University, 6Kinmen Hospital, Ministry of Health and Welfare

This protocol integrated near-infrared spectroscopy into conventional cardiopulmonary exercise testing to identify the involvement of the cerebral hemodynamic response in exercise intolerance in patients with heart failure.

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JoVE Core

A Spine Robotic-Assisted Navigation System for Pedicle Screw Placement
Hsuan-Yu Chen 1,2,3, Xiu-Yun Xiao 4, Chih-Wei Chen 1,4, Hao-Kai Chou 1,4, Chen-Yu Sung 4, Feng Huei Lin 1, Po-Quang Chen 2, Tze-hong Wong 2
1Institute of Biomedical Engineering, National Taiwan University, 2Department of Orthopedic Surgery, National Taiwan University Hospital, Hsin-Chu Branch, 3Department of Orthopedic Surgery, National Taiwan University Hospital, Hsin-Chu Biomedical Park Branch, 4Point Robotics MedTech Inc.

This article presents a standardized surgical technique for robotic-assisted pedicle screw placement by using robotic-assisted navigational systems. We present a step-by-step protocol and describe the workflow and precautions of this procedure.

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Bioengineering

A Custom Multiphoton Microscopy Platform for Live Imaging of Mouse Cornea and Conjunctiva
Yueh-Feng Wu 1, Rai-Teng Ye 1, Ming-Kai Pan 2,3, Sung-Jan Lin *1,3,4,5, Hsin-Yuan Tan *6,7
1Department of Biomedical Engineering, National Taiwan University, 2Institute of Pharmacology, College of Medicine, National Taiwan University, 3Molecular Imaging Center, National Taiwan University, 4Department of Dermatology, National Taiwan University Hospital, and College of Medicine, 5Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, 6Department of Ophthalmology, Chang Gung Memorial Hospital, 7College of Medicine, Chang Gung University

Presented here is a multiphoton microscopic platform for live mouse ocular surface imaging. Fluorescent transgenic mouse enables the visualization of cell nuclei, cell membranes, nerve fibers and capillaries within the ocular surface. Non-linear second harmonic generation signals derived from collagenous structures provide label-free imaging for stromal architectures.

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Medicine

Model of Ischemic Heart Disease and Video-Based Comparison of Cardiomyocyte Contraction Using hiPSC-Derived Cardiomyocytes
Yun Liu *1, Yin Liang *1, Mengxue Wang *1, Chen Wang 1, Heng Wei 2, Keiji Naruse 1, Ken Takahashi 1
1Department of Cardiovascular Physiology, Graduate School of Medicine, Dentistry and Pharmaceutical Sciences, Okayama University, 2Institute of Laboratory Animals, Graduate School of Medicine, Kyoto University

We present a model of ischemic heart disease using cardiomyocytes derived from human induced pluripotent stem cells, together with a method for quantitative evaluation of tissue damage caused by ischemia. This model can provide a useful platform for drug screening and further research on ischemic heart disease.

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Medicine

A Preclinical Controlled Cortical Impact Model for Traumatic Hemorrhage Contusion and Neuroinflammation
Hung-Fu Lee *1, Chih Hung Chen *2, Che-Feng Chang 2
1Department of Neurosurgery, Cheng Hsin General Hospital, 2Graduate Institute of Physiology, College of Medicine, National Taiwan University

Intraparenchymal hemorrhage and neuroinflammation accompanied by cerebral contusion can trigger severe secondary brain injury. This protocol details a mouse controlled cortical impact (CCI) model allowing researchers to study hemorrhage contusion and post-traumatic immune responses and explore potential therapeutics.

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Neuroscience

Using Baseplating and a Miniscope Preanchored with an Objective Lens for Calcium Transient Research in Mice
Yi-Tse Hsiao 1, Angela Yu-Chi Wang 1, Ting-Yen Lee 1, Ching-Yuan Chang 1
1Department of Veterinary Medicine, School of Veterinary Medicine, National Taiwan University

The shrinkage of dental cement during curing displaces the baseplate. This protocol minimizes the problem by creating an initial foundation of the dental cement that leaves space to cement the baseplate. Weeks later, the baseplate can be cemented in position on this scaffold using little new cement, therebyreducing shrinkage.

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Biochemistry

Preparation of High-Temperature Sample Grids for Cryo-EM
Yuan-Chih Chang 1,2, Chin-Yu Chen 3,4, Ming-Daw Tsai 1,5
1Institute of Biological Chemistry, Academia Sinica, 2Academia Sinica Cryo-EM Facility, Academia Sinica, 3Department of Life Sciences, National Central University, 4Hubei University, 5Institute of Biochemical Sciences, National Taiwan University

This paper provides a detailed protocol for preparing sample grids at temperatures as high as 70 °C, prior to plunge freezing for cryo-EM experiments.

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Biochemistry

Uracil-DNA Glycosylase Assay by Matrix-assisted Laser Desorption/Ionization Time-of-flight Mass Spectrometry Analysis
Hui-Lan Chang *1, Kang-Yi Su *1,2, Steven D. Goodman 3, Wern-Cherng Cheng 2, Liang-In Lin 1,2, Ya-Chien Yang 1,2, Sui-Yuan Chang 1,2, Woei-horng Fang 1,2
1Department of Clinical Laboratory Sciences and Medical Biotechnology, College of Medicine, National Taiwan University, 2Department of Laboratory Medicine, National Taiwan University Hospital, 3Center for Microbial Pathogenesis, Nationwide Children’s Hospital and the Department of Pediatrics, The Ohio State University

A non-labeled, non-radio-isotopic method to assay uracil-DNA glycosylase activity was developed using MALDI-TOF mass spectrometry for direct apurinic/apyrimidinic site-containing product analysis. The assay proved to be quite simple, specific, rapid, and easy to use for DNA glycosylase measurement.

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Environment

An Application for Pairing with Wearable Devices to Monitor Personal Health Status
Yu-Chen Yeh 1, Ang Yeh 1, Shih-Han Hung 1, Chia-Ching Wu 2, Yu-Hsin Tung 1, Sung-Yueh Liu 1, William C. Sullivan 2,3, Chun-Yen Chang 1
1Department of Horticulture and Landscape Architecture, National Taiwan University, 2Department of Landscape Architecture, University of Illinois, Champaign, 3Smart, Healthy Community Initiative, Office of the Provost, University of Illinois, Champaign

The present protocol introduces a non-commercial self-developed application for collecting real-time on-site data, including psychological scales, GPS location, heart rate, and blood-oxygen saturation level, as well as the application's operating procedures. An empirical study conducted in Taiwan in 2020 was used as an application example.

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Medicine

High-Throughput Optical Controlling and Recording Calcium Signal in iPSC-Derived Cardiomyocytes for Toxicity Testing and Phenotypic Drug Screening
Yu-Fen Chang 1, Wan-Chi Su 1, Chih-Chuan Su 1, Min-Wen Chung 1, Jin Chang 2, You-Yi Li 3, Yi-Ju Kao 3, Wen-Pin Chen 3, Matthew J. Daniels 4,5,6
1LumiSTAR Biotechnology, Inc., 2NEXEL Co., Ltd., 3Institute of Pharmacology, College of Medicine, National Taiwan University, 4Manchester Heart Centre, Manchester Royal Infirmary, Manchester University NHS Foundation Trust, 5Division of Cardiovascular Sciences, Manchester Academic Health Sciences Centre, University of Manchester, 6Division of Cell Matrix Biology and Regenerative Medicine, University of Manchester

The present protocol describes all-optical control and observation of triggered cellular activity in iPSC-derived cardiomyocytes (iPSC-CMs) for high throughput drug screening and toxicity testing. Multi-parametric quantification of phenotypic patterns in time, and space, are shown. Long-term effects of drugs over hours, or sequential measurements over days, are demonstrated.

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Biology

Isolation and Characterization of Intact Phycobilisome in Cyanobacteria
Han-Wei Jiang 1, Ming-Yang Ho 1,2
1Department of Life Science, National Taiwan University, 2Institute of Plant Biology, National Taiwan University

The current protocol details the isolation of phycobilisomes from cyanobacteria by centrifugation through a discontinuous sucrose density gradient. The fractions of intact phycobilisomes are confirmed by 77K fluorescent emission spectrum and SDS-PAGE analysis. The resulting phycobilisome fractions are suitable for negative staining of TEM and mass spectrometry analysis.

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Biology

Monitoring Stub1-Mediated Pexophagy
Bo-Hua Chen 1, Wei Yuan Yang 1,2
1Institute of Biological Chemistry, Academia Sinica, 2Institute of Biochemical Sciences, College of Life Sciences, National Taiwan University

This protocol provides instructions for triggering and monitoring Stub1-mediated pexophagy in live cells.

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Biology

Development of a Cabbage Protoplast System for Studying Hypoxia Tolerance in Brassica
Shih-Jie Huang 1, Fu-Chiun Hsu 1
1Department of Horticulture and Landscape Architecture, National Taiwan University

This protocol describes an efficient cabbage mesophyll protoplast system. Various oxygen-deficient treatments were tested, and the system showed a high activation of hypoxia-responsive genes, facilitating studies of the genetic and molecular mechanisms of flooding tolerance in Brassicaceae vegetables.

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Biology

Translation Efficiency Test Using Polysome Profiles Under Heat Stress
Hui-Hsien Chang 1, Mei-Chun Cheng 1
1Department of Biochemical Science and Technology, National Taiwan University

The protocol presented here involves polysomal profiling to isolate translatome, mRNAs associated with ribosomes, into non-polysomal and polysomal RNAs from Arabidopsis through sucrose density gradient centrifugation. This method demonstrates the translation efficiency of heat-stressed Arabidopsis.

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