National Taiwan University 43 articles published in JoVE Biology Monitoring Stub1-Mediated Pexophagy Bo-Hua Chen1, Wei Yuan Yang1,2 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. 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. Goodman3, Wern-Cherng Cheng2, Liang-In Lin1,2, Ya-Chien Yang1,2, Sui-Yuan Chang1,2, Woei-horng Fang1,2 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. Medicine High-Throughput Optical Controlling and Recording Calcium Signal in iPSC-Derived Cardiomyocytes for Toxicity Testing and Phenotypic Drug Screening Yu-Fen Chang1, Wan-Chi Su1, Chih-Chuan Su1, Min-Wen Chung1, Jin Chang2, You-Yi Li3, Yi-Ju Kao3, Wen-Pin Chen3, Matthew J. Daniels4,5,6 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. Environment An Application for Pairing with Wearable Devices to Monitor Personal Health Status Yu-Chen Yeh1, Ang Yeh1, Shih-Han Hung1, Chia-Ching Wu2, Yu-Hsin Tung1, Sung-Yueh Liu1, William C. Sullivan2,3, Chun-Yen Chang1 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. Biology Isolation and Characterization of Intact Phycobilisome in Cyanobacteria Han-Wei Jiang1, Ming-Yang Ho1,2 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. Biochemistry Preparation of High-Temperature Sample Grids for Cryo-EM Yuan-Chih Chang1,2, Chin-Yu Chen3,4, Ming-Daw Tsai1,5 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. Neuroscience Using Baseplating and a Miniscope Preanchored with an Objective Lens for Calcium Transient Research in Mice Yi-Tse Hsiao1, Angela Yu-Chi Wang1, Ting-Yen Lee1, Ching-Yuan Chang1 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. Bioengineering A Custom Multiphoton Microscopy Platform for Live Imaging of Mouse Cornea and Conjunctiva Yueh-Feng Wu1, Rai-Teng Ye1, Ming-Kai Pan2,3, 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. Bioengineering A Spine Robotic-Assisted Navigation System for Pedicle Screw Placement Hsuan-Yu Chen1,2,3, Xiu-Yun Xiao4, Chih-Wei Chen1,4, Hao-Kai Chou1,4, Chen-Yu Sung4, Feng Huei Lin1, Po-Quang Chen2, Tze-hong Wong2 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. Behavior Examining Online Syntactic Processing of Spoken Complex Sentences in Chinese Using Dual-Modal Interference Tasks Tuyuan Cheng1, Jei-Tun Wu2 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. Environment Standard Operating Procedure for Lyssavirus Surveillance of the Bat Population in Taiwan Wei-Cheng Hsu1, Chao-Lung Hsu2,3, Yang-Chang Tu1, Jen-Chieh Chang1, Kwok-Rong Tsai1, Fan Lee1, Shu-Chia Hu1 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. Behavior Computerized Adaptive Testing System of Functional Assessment of Stroke Gong-Hong Lin1, Yi-Jing Huang1, Yeh-Tai Chou2, Hsin-Yu Chiang3, Ching-Lin Hsieh1,4,5 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. Chemistry Extending the Lifespan of Soluble Lead Flow Batteries with a Sodium Acetate Additive Yan-Ting Lin1, Wei-Cheng Kuo1, Chun-Yen Lee1, Hao-Lun Tan1, Hsun-Yi Chen1, Ho-Wei Chan1, Yong-Hong Lai1, Kai-Rui Pan1 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. Environment Use of Autometallography to Localize and Semi-Quantify Silver in Cetacean Tissues Wen-Ta Li1, Bang-Yeh Liou1, Wei-Cheng Yang2, Meng-Hsien Chen3, Hui-Wen Chang1,2, Hue-Ying Chiou4, Victor Fei Pang1,2, Chian-Ren Jeng1,2 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. Chemistry An Efficient Sample Preparation Method to Enhance Carbohydrate Ion Signals in Matrix-assisted Laser Desorption/Ionization Mass Spectrometry Yu-Meng Ou1,2, Shu-Yun Kuo1, Hsun Lee1, Huan-Tsung Chang2, Yi-Sheng Wang1 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. Biochemistry 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 Lai1, Rong-Syuan Yen1, Wei-Yao Hu1, Wern-Cherng Cheng2, Liang-In Lin1,2, Ya-Chien Yang1,2, Woei-Horng Fang1,2 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. Environment A Novel Method for the Pentosan Analysis Present in Jute Biomass and Its Conversion into Sugar Monomers Using Acidic Ionic Liquid Babasaheb M. Matsagar1, Shahriar A. Hossain2,3, Tofazzal Islam4, Yusuke Yamauchi2,3,5,6, Kevin C.-W. Wu1 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). 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. Biology Practical Use of RNA Interference: Oral Delivery of Double-stranded RNA in Liposome Carriers for Cockroaches Jia-Hsin Huang1, Yun Liu1, Yu-Hsien Lin1,2,3, Xavier Belles4, How-Jing Lee1 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. Engineering Well-aligned Vertically Oriented ZnO Nanorod Arrays and their Application in Inverted Small Molecule Solar Cells Ming-Yi Lin1, Shang-Hsuan Wu2, Li-Jen Hsiao3, Widhya Budiawan2, Shih-Lun Chen4, Wei-Chen Tu4, Chia-Yen Lee1, Yia-Chung Chang2, Chih-Wei Chu2 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%. Environment An Ultra-clean Multilayer Apparatus for Collecting Size Fractionated Marine Plankton and Suspended Particles Liang-Saw Wen1, Chih-Ping Lee1, Wen-Huei Lee1, Alan Chuang2 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. Genetics Transient Expression of Foreign Genes in Insect Cells (sf9) for Protein Functional Assay Ju-Chun Chang1, Se Jin Lee2, Jae Su Kim2, Chung-Hsiung Wang3, Yu-Shin Nai1 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. Biology 3D Mitochondrial Ultrastructure of Drosophila Indirect Flight Muscle Revealed by Serial-section Electron Tomography Yi-fan Jiang1, Hsiang-ling Lin2, Chi-yu Fu2 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. Engineering Preparation of Large-area Vertical 2D Crystal Hetero-structures Through the Sulfurization of Transition Metal Films for Device Fabrication Chong-Rong Wu1,2, Tung-Wei Chu2,3, Kuan-Chao Chen1,2, Shih-Yen Lin1,2 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. Bioengineering An Integrated System to Remotely Trigger Intracellular Signal Transduction by Upconversion Nanoparticle-mediated Kinase Photoactivation Hua-De Gao1,2, Pounraj Thanasekaran1, Tzu-Ho Chen1,2, Yu-Hsu Chang3, Yu-Ju Chen1,2, Hsien-Ming Lee1 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. Bioengineering Magnetic and Thermal-sensitive Poly(N-isopropylacrylamide)-based Microgels for Magnetically Triggered Controlled Release Chih-Yu Kuo1, Ting-Yu Liu2, Kuan-Syun Wang2, Andri Hardiansyah3, Yen-Ting Lin4, Hsueh-Yung Chen4, Wen-Yen Chiu1,4,5 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. Engineering Preparation of Janus Particles and Alternating Current Electrokinetic Measurements with a Rapidly Fabricated Indium Tin Oxide Electrode Array Yu-Liang Chen1, Hong-Ren Jiang1 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. Engineering Pool-Boiling Heat-Transfer Enhancement on Cylindrical Surfaces with Hybrid Wettable Patterns Sujith Kumar C.S.*1, Yao Wen Chang*1, Ping-Hei Chen1 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. Behavior Manipulation of Epileptiform Electrocorticograms (ECoGs) and Sleep in Rats and Mice by Acupuncture Pei-Lu Yi*1, Shuo-Bin Jou*2, Yi-Jou Wu3, Fang-Chia Chang3,4,5 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. 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 Tzeng1 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. Medicine A Model of Cardiac Remodeling Through Constriction of the Abdominal Aorta in Rats Hui-Chun Ku1, Shih-Yi Lee2,3, Yuan-Kun Aden Wu4, Kai-Chien Yang1,5, Ming-Jai Su1 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. Biochemistry Preparation of Homogeneous MALDI Samples for Quantitative Applications Yu-Meng Ou1,2, Chien-Wei Tsao1,3, Yin-Hung Lai1, Hsun Lee1, Huan-Tsung Chang2, Yi-Sheng Wang1 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. Genetics The Visual Colorimetric Detection of Multi-nucleotide Polymorphisms on a Pneumatic Droplet Manipulation Platform Szu-I Yeh1, Wei-Feng Fang1, Chao-Jyun Huang1, Tzu-Ming Wang1, Jing-Tang Yang1 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. Developmental Biology In Vitro and In Vivo Models to Study Corneal Endothelial-mesenchymal Transition Wei-Ting Ho1,2, Chien-Chia Su3, Jung-Shen Chang3, Shu-Wen Chang1, Fung-Rong Hu3, Tzuu-Shuh Jou2,4, I-Jong Wang3,4 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. Bioengineering Designing Microfluidic Devices for Studying Cellular Responses Under Single or Coexisting Chemical/Electrical/Shear Stress Stimuli Tzu-Yuan Chou1, Yung-Shin Sun2, Hsien-San Hou3, Shang-Ying Wu1, Yun Zhu1, Ji-Yen Cheng3, Kai-Yin Lo1 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. 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. Santschi4 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. 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 Wu1 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. Developmental Biology Isolation and Differentiation of Adipose-Derived Stem Cells from Porcine Subcutaneous Adipose Tissues Yu-Jen Chen1,2, Hui-Yu Liu2, Yun-Tsui Chang2, Ying-Hung Cheng2, Harry J. Mersmann2, Wen-Hung Kuo3, Shih-Torng Ding1,2 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. Developmental Biology Primary Endodermal Epithelial Cell Culture from the Yolk Sac Membrane of Japanese Quail Embryos Han Jen Lin1, Siou Huei Wang1, Yu Hui Pan1, Shih-Torng Ding1 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. Developmental Biology Identification of Critical Conditions for Immunostaining in the Pea Aphid Embryos: Increasing Tissue Permeability and Decreasing Background Staining Gee-Way Lin1,2, Chun-che Chang1,2,3,4 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. Engineering A Fabrication Method for Highly Stretchable Conductors with Silver Nanowires Chia-Wei Chang1, Shih-Pin Chen1, Ying-Chih Liao1 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. Engineering Measurement of Scattering Nonlinearities from a Single Plasmonic Nanoparticle Hsuan Lee1, Kuan-Yu Li1, Yen-Ta Huang1, Po-Ting Shen1, Gitanjal Deka1, Ryosuke Oketani2, Yasuo Yonemaru2, Masahito Yamanaka2, Katsumasa Fujita2, Shi-Wei Chu1,3 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. Bioengineering Analyzing Mixing Inhomogeneity in a Microfluidic Device by Microscale Schlieren Technique Chen-li Sun1, Tzu-hsun Hsiao2 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.
