Chinese Academy of Sciences View Institution's Website 62 articles published in JoVE Biology A Magnetic Separation-Assisted High-Speed Homogenization Method for Large-Scale Production of Endosome-Derived Vesicles Dujuan Wang1,2, Shili Yao2,3, Peng Guo1,2,3,4 1Academy of Medical Engineering and Translational Medicine, Tianjin University, 2Hangzhou Institute of Medicine (HIM), Chinese Academy of Sciences, 3School of Materials Science and Engineering, Tianjin University, 4Zhejiang Cancer Hospital Here, we describe a magnetic separation-assisted high-speed homogenization method for large-scale production of endosome-derived nanovesicles as a new type of exosome mimics (EMs) that share the same biological origin and similar structure, morphology, and protein composition of native extracellular vesicles (EVs). Chemistry Multimodal Nonlinear Hyperspectral Chemical Imaging Using Line-Scanning Vibrational Sum-Frequency Generation Microscopy Jackson C. Wagner1, Bin Yang1,2, Zishan Wu1, Wei Xiong1,3,4 1Department of Chemistry and Biochemistry, UC San Diego, 2State Key Laboratory of Molecular Reaction Dynamics, Dalian Institute of Chemical Physics, Chinese Academy of Sciences, 3Materials Science and Engineering Program, UC San Diego, 4Department of Electrical and Computer Engineering, UC San Diego A multimodal, rapid hyperspectral imaging framework was developed to obtain broadband vibrational sum-frequency generation (VSFG) images, along with brightfield, second harmonic generation (SHG) imaging modalities. Due to the infrared frequency being resonant with molecular vibrations, microscopic structural and mesoscopic morphology knowledge is revealed of symmetry-allowed samples. Biology Optimizing the Rearing Procedure of Germ-Free Wasps Zhengyu Zhu*1,2, Dandan Wang*1, Yanjun Liu1, Ting Tang2, Guan-Hong Wang1 1State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences, 2College of Life Sciences, Hebei University Nasonia wasp embryos were dissected from Lucillia sericata pupae after parasitization for 12-24 h and washed with alcohol and 10% sodium hypochlorite solution to obtain germ-free embryos. After rearing the germ-free embryos and supplying them with Nasonia rearing medium to grow and develop in vitro, germ-free Nasonia adults were obtained. Chemistry Tuning Oxide Properties by Oxygen Vacancy Control During Growth and Annealing Tristan Steegemans*1, Shinhee Yun*1, Carlos N. Lobato1, Eric Brand1, Yunzhong Chen2, Felix Trier1, Dennis V. Christensen1 1Department of Energy Conversion and Storage, Technical University of Denmark, 2Beijing National Laboratory for Condensed Matter Physics and Institute of Physics, Chinese Academy of Sciences Oxide materials show many exotic properties that can be controlled by tuning the oxygen content. Here, we demonstrate the tuning of oxygen content in oxides by varying the pulsed laser deposition parameters and by performing postannealing. As an example, electronic properties of SrTiO3-based heterostructures are tuned by growth modifications and annealing. Environment Collection of Alfalfa Root Exudates to Study the Impact of Di(2-ethylhexyl) Phthalate on Metabolite Production Wenjie Ren1,2, Rui Zhao1, Ying Teng1, Yongming Luo1 1Key Laboratory of Soil Environment and Pollution Remediation, Institute of Soil Science, Chinese Academy of Sciences, 2College of Resources and Environment, University of Chinese Academy of Sciences The secretion of root exudates is usually an external detoxification strategy for plants under stress conditions. This protocol describes how to assess the impact of xenobiotics on alfalfa via nontargeted metabolomic analysis. Biology A Precise and Quantifiable Method for Collecting Hemolymph from Small Arthropods Qing Liu1,3, Lili Zhang1,2, Rongxiang Fang1,2,3, Yan Huo1,2 1State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, 2CAS Center for Excellence in Biotic Interactions, University of Chinese Academy of Sciences, 3University of Chinese Academy of Sciences We describe a method to collect quantifiable hemolymph efficiently from small arthropods for subsequent analysis. Biology Induction and Diverse Assessment Indicators of Experimental Autoimmune Encephalomyelitis Chun Wang*1, Jie Lv*1, Wei Zhuang1,2, Ling Xie1, Guangyu Liu1, Kaidireya Saimaier1, Sanxing Han1, Changjie Shi1, Qiuhong Hua1, Ru Zhang1, Guangfeng Shi3, Changsheng Du1 1Putuo People's Hospital, Shanghai Key Laboratory of Signaling and Disease Research, School of Life Sciences and Technology, Tongji University, 2National Laboratory of Biomacromolecules, CAS Center for Excellence in Biomacromolecules, Institute of Biophysics, Chinese Academy of Sciences, 3Department of Infectious Diseases, Shanghai Key Laboratory of Infectious Diseases and Biosafety Emergency Response, National Medical Center for Infectious Diseases, Huashan Hospital, Fudan University The present protocol describes the induction of experimental autoimmune encephalomyelitis in a mouse model using myelin oligodendrocyte glycoprotein and monitoring the disease process using a clinical scoring system. Experimental autoimmune encephalomyelitis-related symptoms are analyzed using mouse femur micro-computed tomography analysis and open field test to assess the disease process comprehensively. Developmental Biology An Efficient Clearing Protocol for the Study of Seed Development in Tomato (Solanum lycopersicum L.) Yixuan Feng1,2, Tai Wang1,2,3, Lingtong Liu1 1Key Laboratory of Plant Molecular Physiology, Institute of Botany, Chinese Academy of Sciences, 2College of Life Science, University of Chinese Academy of Sciences, 3Innovative Academy of Seed Design, Chinese Academy of Sciences The tomato seed is an important model for studying genetics and developmental biology during plant reproduction. This protocol is useful for clearing tomato seeds at different developmental stages to observe the finer embryonic structure. Bioengineering Cellular Affinity of Particle-Stabilized Emulsion to Boost Antigen Internalization Fengqiang Cao*1,2,3, Yali Ming*3,4, Weixiang Gao3,4, Jia Ge3, Kenji Ogino1,2 1Graduate School of Bio-Applications and Systems Engineering, Tokyo University of Agriculture and Technology, 2Institute of Global Innovation Research, Tokyo University of Agriculture and Technology, 3State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences, 4University of Chinese Academy of Sciences To rationally design efficient adjuvants, we developed poly-lactic-co-glycolic acid nanoparticle-stabilized Pickering emulsion (PNPE). The PNPE possessed unique softness and a hydrophobic interface for potent cellular contact and offered high-content antigen loading, improving the cellular affinity of the delivery system to antigen-presenting cells and inducing efficient internalization of antigens. Biology Mosquito-Associated Virus Isolation from Field-Collected Mosquitoes Doudou Huang*1, Haixia Ma*1, Lu Zhao1,2, Xiaoyu Wang1,2, Yi Huang1,2, Fei Wang1, Zhiming Yuan1,2, Han Xia1,2 1Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences Numerous novel virus-like sequences have been found in mosquitoes due to the extensive use of sequencing technologies. We provide an effective procedure for isolating and amplifying viruses using vertebrate and mosquito cell lines, which might serve as the basis for future studies on mosquito-associated viruses, including mosquito-borne and mosquito-specific viruses. Medicine Establishment of a Simple and Effective Rat Model for Intraoperative Parathyroid Gland Imaging Fan Chen*1,2, Chun Liu*3, Peng Guo3, Weihui Zheng1,2 1Key Laboratory of Head & Neck Cancer Translational Research of Zhejiang Province, Zhejiang Cancer Hospital, 2The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), 3Institute of Basic Medicine and Cancer (IBMC), Chinese Academy of Sciences To date, the development of parathyroid gland (PG) identification methods is limited by the lack of animal models in preclinical research. Here, we establish a simple and effective rat model for intraoperative PG imaging and evaluate its effectiveness by using iron oxide nanoparticles as a novel PG contrast agent. Immunology and Infection Experimental Viral Infection in Adult Mosquitoes by Oral Feeding and Microinjection Fei Wang*1, Cihan Yang*1,2, Shunlong Wang1,2, Qun Wu1,2, Christabel Ochieng1,2, Zhiming Yuan1,2, Han Xia1,2 1Key Laboratory of Special Pathogens and Biosafety, Wuhan Institute of Virology, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences This methodology, which included oral feeding and intrathoracic injection infection, could effectively assess the influence of midgut and/or salivary gland barriers on arbovirus infection. Engineering Rapid Repetition Rate Fluctuation Measurement of Soliton Crystals in a Microresonator Peng Xie*1,2, Xinyu Wang*1,2, Weiqiang Wang1,2, Wenfu Zhang1,2, Zhizhou Lu1,2, Yang Wang1,2, Wei Zhao1,2 1State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics (XIOPM), Chinese Academy of Sciences (CAS), 2University of Chinese Academy of Sciences Here, we present a protocol to generate soliton crystals in a butterfly-packaged micro-ring resonator using a thermal tuned method. Further, the repetition rate fluctuations of a soliton crystal with a single vacancy are measured using a delayed self-heterodyne method. Biochemistry Evaluating the Effect of Pesticides on the Larvae of the Solitary Bees Yingying Song1, Ruiping Li1,2, Lili Li1, Fang Ouyang3, Xingyuan Men1 1Institute of Plant Protection, Shandong Academy of Agricultural Sciences, 2College of Plant Protection, Shandong Agricultural University, 3State Key Laboratory of Integrated Management of Pest Insects and Rodents, Institute of Zoology, Chinese Academy of Sciences The present protocol explains a method to feed pesticide-contaminated provisions to the larvae of the solitary bees, Osmia excavata. The procedure examines the ecotoxicity of the pesticide to the larvae of the solitary bees. Biology Two-layered Membrane Sandwich Method for Laodelphax striatellus Saliva Collection Jing Zhao1,2,3, Jie Yang1,2,3, Lili Zhang1,2, Rongxiang Fang1,2, Yan Huo1,2 1State Key Laboratory of Plant Genomics, Institute of Microbiology, Chinese Academy of Sciences, 2National Plant Gene Research Center, 3University of the Chinese Academy of Sciences The present protocol describes a method to collect sufficient saliva from piercing-sucking insects using an artificial medium. This is a convenient method for collecting insect saliva and studying salivary function on insect feeding behavior and vector-borne virus transmission. Environment Ecotoxicological Effects of Microplastics on Bird Embryo Development by Hatching without Eggshell Liyi Wang1,2, Nana Xue1,2, Wenfeng Li2, Rehemanjiang Wufuer1, Daoyong Zhang1,3 1Xinjiang Key Laboratory of Environmental Pollution and Bioremediation, Xinjiang Institute of Ecology and Geography, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3Key Laboratory of Microbial Technology for Industrial Pollution Control of Zhejiang Province, College of Environment, Zhejiang University of Technology This paper introduces a method of hatching without using an eggshell for toxicological studies of particle pollutants such as microplastics. Developmental Biology Separation of Follicular Cells and Oocytes in Ovarian Follicles of Zebrafish Wenyi Wang1, Tao Kang1, Lin Bai1, Wei Hu2, Yayoi Obata3, Jianzhen Li1 1College of Life Sciences, Northwest Normal University, 2State Key Laboratory of Freshwater Ecology and Biotechnology, Institute of Hydrobiology, Chinese Academy of Sciences, 3Department of Bioscience, Tokyo University of Agriculture Here, we present a simple method for separating follicular cells and oocytes in zebrafish ovarian follicles, which will facilitate investigations of ovarian development in zebrafish. Biology Two-Step Reverse Transcription Droplet Digital PCR Protocols for SARS-CoV-2 Detection and Quantification Raphael Nyaruaba1,2,3, Xiaohong Li1, Caroline Mwaliko2,3,4, Changchang Li1, Matilu Mwau5, Nelson Odiwour1,2,3, Elishiba Muturi1,2,3, Caroline Muema1,2,3, Junhua Li1, Junping Yu1, Hongping Wei1 1Key Laboratory of Special Pathogens and Biosafety, Center for Biosafety Mega-Science, Wuhan Institute of Virology, Chinese Academy of Sciences, 2International College, University of Chinese Academy of Sciences, 3Sino-Africa Joint Research Center, 4CAS Key Laboratory of Molecular Virology & Immunology, Institut Pasteur of Shanghai, Chinese Academy of Sciences, 5Center for Infectious and Parasitic Diseases Control Research, Kenya Medical Research Institute This work summarizes steps on developing different assays for SARS-CoV-2 detection using a two color ddPCR system. The steps are elaborate and notes have been included on how to improve the assays and experiment performance. These assays may be used for multiple SARS-CoV-2 RT-ddPCR applications. Immunology and Infection Stability and Structure of Bat Major Histocompatibility Complex Class I with Heterologous β2-Microglobulin Di Zhang*1,2, Kefang Liu*3,4, Dan Lu*2,5, Pengyan Wang1,2, Qingxu Zhang1,2, Peipei Liu2, Yingze Zhao2, Yan Chai4, Jianxin Lyu1, Jianxun Qi4, William J. Liu1,2 1School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 2NHC Key Laboratory of Biosafety, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 3Faculty of Health Sciences, University of Macau, 4CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, 5Savaid Medical School, University of Chinese Academy of Sciences The protocol describes experimental methods to obtain stable major histocompatibility complex (MHC) class I through potential β2-microglobulin (β2m) substitutions from different species. The structural comparison of MHC I stabilized by homologous and heterologous β2m were investigated. Engineering Improving the Combustion Performance of a Hybrid Rocket Engine using a Novel Fuel Grain with a Nested Helical Structure Zezhong Wang1,2, Xin Lin1, Fei Li1, Zelin Zhang1, Xilong Yu1,2 1Institute of Mechanics, Chinese Academy of Sciences, 2School of Engineering Science, University of Chinese Academy of Sciences A technique utilizing a solid fuel grain with a novel nested helical structure to improve the combustion performance of a hybrid rocket engine is presented. Biochemistry Phosphoproteomic Strategy for Profiling Osmotic Stress Signaling in Arabidopsis Chuan-Chih Hsu1,3, Chia-Feng Tsai2, W. Andy Tao3,4, Pengcheng Wang5 1Department of Plant Biology, Carnegie Institute for Science, 2Graduate School of Pharmaceutical Sciences, Kyoto University, 3Department of Biochemistry, Purdue University, 4Department of Chemistry, Purdue University, 5Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, Chinese Academy of Sciences Presented here is a phosphoproteomic approach, namely stop and go extraction tip based phosphoproteomic, which provides high-throughput and deep coverage of Arabidopsis phosphoproteome. This approach delineates the overview of osmotic stress signaling in Arabidopsis. Engineering Graphene-Assisted Quasi-van der Waals Epitaxy of AlN Film on Nano-Patterned Sapphire Substrate for Ultraviolet Light Emitting Diodes Xiang Zhang*1,2, Zhaolong Chen*3, Hongliang Chang1,2, Jianchang Yan1,2, Shenyuan Yang2,4, Junxi Wang1,2, Peng Gao5, Tongbo Wei1,2 1State Key Laboratory of Solid-State Lighting, Institute of Semiconductors, Chinese Academy of Sciences, 2Center of Materials Science and Optoelectronics Engineering, University of Chinese Academy of Science, 3Center for Nanochemistry (CNC), Beijing National Laboratory for Molecular Science, College of Chemistry and Molecular Engineering, Peking University, 4State Key Laboratory of Superlattices and Microstructures, Institute of Semiconductors, Chinese Academy of Sciences, 5Electron Microscopy Laboratory, School of Physics, Peking University A protocol for graphene-assisted growth of high-quality AlN films on nano-patterned sapphire substrate is presented. Genetics Tomato Root Transformation Followed by Inoculation with Ralstonia Solanacearum for Straightforward Genetic Analysis of Bacterial Wilt Disease Rafael J. L. Morcillo1, Achen Zhao1,2, María I. Tamayo-Navarrete3, José M. García-Garrido3, Alberto P. Macho1 1Shanghai Center for Plant Stress Biology, CAS Center for Excellence in Molecular Plant Sciences, Shanghai Institutes of Biological Sciences, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3Department of Soil Microbiology and Symbiotic Systems, Estación Experimental del Zaidín (CSIC) Here, we present a versatile method for tomato root transformation followed by inoculation with Ralstonia solanacearum to perform straightforward genetic analysis for the study of bacterial wilt disease. Engineering Thermocapillary Convection Space Experiment on the SJ-10 Recoverable Satellite Li Duan*1,2, Yongli Yin*3, Jia Wang*1, Qi Kang1,2, Di Wu1, Huan Jiang1, Pu Zhang1, Liang Hu1 1National Microgravity Laboratory, Institute of Mechanics, Chinese Academy of Sciences, 2School of Engineering Sciences, University of Chinese Academy of Sciences, 3China Astronaut Research and Training Center A protocol for the space payload design, the space experiment on thermocapillary convection, and analyses of experimental data and images are presented in this paper. Developmental Biology Isolation, Culture, and Adipogenic Induction of Neural Crest Original Adipose-Derived Stem Cells from Periaortic Adipose Tissue Yiding Qi*1, Xiang Miao*2, Lian Xu3, Mengxia Fu3, Shi Peng4, Kailei Shi5, Jun Li4, Maoqing Ye5, Ruogu Li1 1Department of Cardiology, Shanghai Chest Hospital, Shanghai Jiao Tong University, 2Shanghai Institute of Nutrition and Health, Shanghai Institute for Biological Sciences, Chinese Academy of Sciences, 3Shanghai Key Laboratory of Hypertension, Shanghai Institute of Hypertension, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, 4Department of Cardiology, Shanghai General Hospital, Shanghai Jiao Tong University School of Medicine, 5Department of Cardiology, Shanghai Key Laboratory of Clinical Geriatric Medicine, Huadong Hospital Affiliated to Fudan University We present a protocol for the isolation, culture, and adipogenic induction of neural crest derived adipose-derived stem cells (NCADSCs) from the periaortic adipose tissue of Wnt-1 Cre+/-;Rosa26RFP/+ mice. The NCADSCs can be an easily accessible source of ADSCs for modeling adipogenesis or lipogenesis in vitro. Neuroscience Stimulation Location Determination using a 3D Digitizer with High-Definition Transcranial Direct Current Stimulation Wanting Chen1, Rui Chen1, Qinghua He1,2,3,4,5 1Faculty of Psychology, Southwest University, 2Key Laboratory of Cognition and Personality, Ministry of Education, Southwest University, 3Southwest University Branch, Collaborative Innovation Center of Assessment toward Basic Education Quality, Beijing Normal University, 4Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences, 5Chongqing Collaborative Innovation Center for Brain Science Presented here is a protocol to achieve higher accuracy in determination of stimulation location combining a 3D digitizer with high-definition transcranial direct current stimulation. Developmental Biology An Enhanced Green Fluorescence Protein-based Assay for Studying Neurite Outgrowth in Primary Neurons Wai Wa Ray Chan*1, Wen Li*1,2, Dik Long Dennis Chau*1, Kwok-Fai Lau1 1School of Life Sciences, The Chinese University of Hong Kong, Hong Kong Special Administrative Region, 2Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, Shenzhen In this report, we describe a simple protocol for studying neurite outgrowth in embryonic rat cortical neurons by co-transfecting with EGFP and the protein of interest. Chemistry Isotopic Effect in Double Proton Transfer Process of Porphycene Investigated by Enhanced QM/MM Method Zhihui Tu1,2, Jian Yin3, Liangxu Xie4 1State Key Laboratory of Structural Chemistry, Fujian Institute of Research on the Structure of Matter, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3XtalPi Inc. (Shenzhen Jingtai Technology Co., Ltd.), 4Institute of Bioinformatics and Medical Engineering, School of Electrical and Information Engineering, Jiangsu University of Technology A protocol that uses enhanced QM/MM method to investigate the isotopic effect on the double proton transfer process in porphycene is presented here. Engineering Quantitative Analysis of Vacuum Induction Melting by Laser-induced Breakdown Spectroscopy Tianzhuo Zhao1,2,3, Xin Li1,2, Qixiu Zhong1,2, Hong Xiao1,3, Shuzhen Nie1,3, Fuqiang Lian1,3, Sining Sun4, Zhongwei Fan1,2,3 1The Academy of Opto-electronics, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3National Engineering Research Center for DPSSL, 4Beijing GK Laser Technology Co., Ltd. During vacuum induction melting, laser-induced breakdown spectroscopy is used to perform real-time quantitative analysis of the main-ingredient elements of a molten alloy. Chemistry Separation of Uranium and Thorium for 230Th-U Dating of Submarine Hydrothermal Sulfides Lisheng Wang1,2,3, Xuefeng Wang1,2, Jun Ye4, Zhibang Ma1,2, Weifang Yang5, Jule Xiao1,2 1Key Laboratory of Cenozoic Geology and Environment, Institute of Geology and Geophysics, Chinese Academy of Sciences, 2Institutions of Earth Science, Chinese Academy of Sciences, 3University of Chinese Academy of Sciences, 4First Institute of Oceanography, State Oceanic Administration, 5Key Laboratory of Submarine Geosciences, Second Institute of Oceanography, State Oceanic Administration The protocol describes a method to purify and separate the U and Th nuclide in submarine hydrothermal sulfide sample with Fe co-precipitation and extraction chromatography for 230Th-U disequilibrium dating. Environment A High-throughput Assay for the Prediction of Chemical Toxicity by Automated Phenotypic Profiling of Caenorhabditis elegans Shan Gao*1, Weiyang Chen*2, Nan Zhang1, Chi Xu3, Haiming Jing1,4, Wenjing Zhang1,4, Gaochao Han1,4, Matthew Flavel5, Markandeya Jois5, Yingxin Zeng1, Jing-Dong J. Han3, Bo Xian3, Guojun Li1,4 1Beijing Key Laboratory of Diagnostic and Traceability Technologies for Food Poisoning, Beijing Center for Disease Prevention and Control/Beijing Center of Preventive Medicine Research, China, 2College of Computer Science and Technology, Qilu University of Technology(Shandong Academy of Sciences), China, 3Key Laboratory of Computational Biology, CAS Center for Excellence in Molecular Cell Science, Collaborative Innovation Center for Genetics and Developmental Biology, Chinese Academy of Sciences-Max Planck Partner Institute for Computational Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, China, 4Beijing Key Laboratory of Environmental Toxicology, School of Public Health, Capital Medical University, China, 5School of Life Sciences, La Trobe University, Australia A quantitative method has been developed to identify and predict the acute toxicity of chemicals by automatically analyzing the phenotypic profiling of Caenorhabditis elegans. This protocol describes how to treat worms with chemicals in a 384-well plate, capture videos, and quantify toxicological related phenotypes. Immunology and Infection Establishment of Viral Infection and Analysis of Host-Virus Interaction in Drosophila Melanogaster Shuo Yang*1,2,3, Yaya Zhao*1,2, Junjing Yu4, Zhiqin Fan2, Si-tang Gong1, Hong Tang1,2, Lei Pan1,2,3 1 This protocol describes how to establish viral infection in vivo in Drosophila melanogaster using the nano-injection method and basic techniques to analyze virus-host interaction. Behavior A Conflict Model of Reward-seeking Behavior in Male Rats Shaofei Jiang1,2, Yue Zhang1,2, Xigeng Zheng1,2, Haoshuang Luo1,2, Zhengkui Liu1,2, Yunjing Bai1,2 1CAS Key Laboratory of Mental Health, Institute of Psychology, Chinese Academy of Sciences, 2Department of Psychology, University of Chinese Academy of Sciences This conflict model is used to measure the impairment of inhibitory control after exposure to addictive drugs, or other factors that may influence inhibitory control. A sexual stimulus and an aversive obstacle are concurrently presented, thus male rats have to conquer the obstacle to approach the sexual reward. Behavior Simultaneous Recordings of Cortical Local Field Potentials and Electrocorticograms in Response to Nociceptive Laser Stimuli from Freely Moving Rats Lupeng Yue1,2, Fengrui Zhang3, Xuejing Lu1,2, You Wan4,5,6, Li Hu1,2,7 1CAS Key Laboratory of Mental Health, Institute of Psychology, 2Department of Psychology, University of Chinese Academy of Sciences, 3Research Center of Brain Cognitive Neuroscience, Liaoning Normal University, 4Neuroscience Research Institute, Peking University, 5Department of Neurobiology, School of Basic Medical Sciences, Peking University, 6Key Laboratory for Neuroscience, Ministry of Education/National Health and Family Planning Commission, Peking University, 7Department of Pain Management, State Key Clinical Specialty in Pain Medicine, Second Affiliated Hospital of Guangzhou Medical University We developed a technique that simultaneously records both electrocorticography and local field potentials in response to nociceptive laser stimuli from freely moving rats. This technique helps establish a direct relationship of electrocortical signals at the mesoscopic and macroscopic levels, which facilitates the investigation of nociceptive information processing in the brain. Engineering Quantitative Analysis by Thermogravimetry-Mass Spectrum Analysis for Reactions with Evolved Gases Rongbin Li1, Qian Huang2, Kai Wei2, Hongde Xia2 1School of Metallurgical and Ecological Engineering, University of Science and Technology Beijing, 2Institute of Engineering Thermophysics, Chinese Academy of Sciences Precise determination of the evolved gases' flow rate is key to study the details of reactions. We provide a novel quantitative analysis method of equivalent characteristic spectrum analysis for thermogravimetry-mass spectrum analysis by establishing the calibration system of the characteristic spectrum and relative sensitivity, for obtaining the flow rate. Immunology and Infection Evaluation of Zika Virus-specific T-cell Responses in Immunoprivileged Organs of Infected Ifnar1-/- Mice Yongli Zhang*1,2, Hangjie Zhang*2, Wenqiang Ma*3, Kefang Liu1,2, Min Zhao4, Yingze Zhao2, Xuancheng Lu5, Fuping Zhang6, Xiangdong Li3, George F. Gao1,2,4,6, William J. Liu1,2 1School of Laboratory Medicine and Life Sciences, Wenzhou Medical University, 2NHC Key Laboratory of Medical Virology and Viral Diseases, National Institute for Viral Disease Control and Prevention, Chinese Center for Disease Control and Prevention, 3State Key Laboratory of Agrobiotechnology, College of Biological Sciences, China Agricultural University, 4Research Network of Immunity and Health (RNIH), Beijing Institutes of Life Science, Chinese Academy of Sciences, 5Laboratory Animal Center, Chinese Center for Disease Control and Prevention, 6CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences A protocol to evaluate antigen-specific T-cell responses in the immunoprivileged organs of the Ifnar1-/- murine model for the Zika virus (ZIKV) infection is described. This method is pivotal for investigating the cellular mechanisms of the protection and immunopathogenesis of ZIKV vaccines and is also valuable for their efficacy evaluation. Developmental Biology A Familial Hypercholesterolemia Human Liver Chimeric Mouse Model Using Induced Pluripotent Stem Cell-derived Hepatocytes Jiayin Yang*1,2,6, Lai-Yung Wong*2, Xiao-Yu Tian*3, Rui Wei1,2, Wing-Hon Lai2, Ka-Wing Au2, Zhiwei Luo4,5, Carl Ward4,5, Wai-In Ho2, David P. Ibañez4,5, Hao Liu4,5, Xichen Bao4,5, Baoming Qin4,5, Yu Huang3, Miguel A. Esteban4,5,7, Hung-Fat Tse1,2,6,7 1Department of Medicine, University of Hong Kong-Shenzhen Hospital, 2The Cardiology Division, Department of Medicine, Li Ka Shing Faculty of Medicine, University of Hong Kong, 3School of Biomedical Sciences, Institute of Vascular Medicine, Li Ka Shing Institute of Health Sciences, Chinese University of Hong Kong, 4Key Laboratory of Regenerative Biology of the Chinese Academy of Sciences, Joint School of Life Sciences, Guangzhou Institutes of Biomedicine and Health and Guangzhou Medical University, 5Laboratory of RNA, Chromatin, and Human Disease, CAS Key Laboratory of Regenerative Biology and Guangdong Provincial Key Laboratory of Stem Cells and Regenerative Medicine, Guangzhou Institutes of Biomedicine and Health, Chinese Academy of Sciences, 6Research Centre of Heart, Brain, Hormone, and Healthy Ageing, Li Ka Shing Faculty of Medicine, University of Hong Kong, 7Hong Kong-Guangdong Stem Cell and Regenerative Medicine Research Centre, University of Hong Kong and Guangzhou Institutes of Biomedicine and Health Here, we present a protocol to generate a human liver chimeric mouse model of familial hypercholesterolemia using human induced pluripotent stem cell-derived hepatocytes. This is a valuable model for testing new therapies for hypercholesterolemia. Bioengineering Comprehensive Evaluation of the Effectiveness and Safety of Placenta-Targeted Drug Delivery Using Three Complementary Methods Baozhen Zhang1, Zhilong Chen1,2, Jinyu Han1,3, Mengxia Li1, Nihar R. Nayak4, Xiujun Fan1 1Laboratory for Reproductive Health, Institute of Biomedicine and Biotechnology, Shenzhen Institutes of Advanced Technology, Chinese Academy of Sciences, 2College of Veterinary Medicine, Hunan Agricultural University, 3Key Laboratory of Chemical Engineering Process and Technology for High-efficiency Conversion, College of Chemistry and Material Sciences, Heilongjiang University, 4Department of Obstetrics and Gynecology, Wayne State University School of Medicine We describe a system that utilizes three methods to evaluate the safety and effectiveness of placenta-targeted drug delivery: in vivo imaging to monitor nanoparticle accumulation, high-frequency ultrasound to monitor placental and fetal development, and HPLC to quantify drug delivery to tissue. Environment Evolution of Staircase Structures in Diffusive Convection Shuang-Xi Guo1, Sheng-Qi Zhou1, Xian-Rong Cen1, Yuan-Zheng Lu1 1State Key Laboratory of Tropical Oceanography, South China Sea Institute of Oceanology, Chinese Academy of Sciences Diffusive convection (DC) widely occurs in natural processes and engineering applications, characterized by a series of staircases with homogeneous convecting layers and stratified interfaces. An experimental procedure is described to simulate the evolution process of the DC staircase structure, including the generation, development and disappearance, in a rectangular tank. Biology Visualizing the Interaction Between the Qdot-labeled Protein and Site-specifically Modified λ DNA at the Single Molecule Level Huijun Xue*1,2, Zhengyan Zhan*1, Kaining Zhang1,2, Yu Vincent Fu1,2 1State Key Laboratory of Microbial Resources, Institute of Microbiology, Chinese Academy of Sciences, 2Savaid Medical School, University of Chinese Academy of Sciences Here, we present a protocol to study DNA-protein interactions by total internal reflection fluorescence microscopy (TIRFM) using a site-specifically modified λ DNA substrate and a Quantum-dot labeled protein. Engineering Generation and Coherent Control of Pulsed Quantum Frequency Combs Benjamin MacLellan*1, Piotr Roztocki*1, Michael Kues1,2, Christian Reimer1, Luis Romero Cortés1, Yanbing Zhang1, Stefania Sciara1,3, Benjamin Wetzel1,4, Alfonso Cino3, Sai T. Chu5, Brent E. Little6, David J. Moss7, Lucia Caspani8, José Azaña1, Roberto Morandotti1,9,10 1Institut National de la Recherche Scientifique - Centre Énergie, Matériaux et Télécommunications (INRS-EMT), 2School of Engineering, University of Glasgow, 3Department of Energy, Information Engineering and Mathematical Models, University of Palermo, 4School of Mathematical and Physical Sciences, University of Sussex, 5Department of Physics and Material Science, City University of Hong Kong, 6State Key Laboratory of Transient Optics and Photonics, Xi'an Institute of Optics and Precision Mechanics, Chinese Academy of Science, 7Centre for Micro Photonics, Swinburne University of Technology, 8Institute of Photonics, Department of Physics, University of Strathclyde, 9Institute of Fundamental and Frontier Sciences, University of Electronic Science and Technology of China, 10National Research University of Information Technologies, Mechanics and Optics A protocol is presented for the practical generation and coherent manipulation of high-dimensional frequency-bin entangled photon states using integrated micro-cavities and standard telecommunications components, respectively. Biochemistry An Alternative Culture Method to Maintain Genomic Hypomethylation of Mouse Embryonic Stem Cells Using MEK Inhibitor PD0325901 and Vitamin C Cuiping Li1, Weiyi Lai1, Hailin Wang1 1State Key Laboratory of Environmental Chemistry and Ecotoxicology, Research Center for Eco-Environmental Sciences, Chinese Academy of Sciences We described in detail two chemical-based protocols for culturing mouse embryonic stem cells. This new method utilizes synergistic mechanisms of promoting Tet-mediated oxidation (by vitamin C) and repressing de novo synthesis of 5-methylcytosine (by PD0325901) to maintain DNA hypomethylation and pluripotency of mouse embryonic stem cells. Behavior Combining Quantitative Food-intake Assays and Forcibly Activating Neurons to Study Appetite in Drosophila Lifen Jiang*1,2, Yinpeng Zhan*2,3, Yan Zhu2,3 1School of Life Science, University of Science and Technology of China, 2State key Laboratory of Brain and Cognitive Science, Institute of Biophysics, Chinese Academy of Sciences, 3University of Chinese Academy of Sciences Quantitative food-intake assays with dyed food provide a robust and high-throughput means to evaluate feeding motivation. Combining the food consumption assay with thermogenetic and optogenetic screens is a powerful approach to investigate the neural circuits underlying appetite in adult Drosophila melanogaster. Immunology and Infection An Efficient and Simple Method to Establish NK and T Cell Lines from Patients with Chronic Active Epstein-Barr Virus Infection Chongfeng Xu1, Junhong Ai2, Qingxun Zhang1, Ting Li3, Xiaorong Wu1, Zhengde Xie2, Ziyuan Duan1 1Genetic Resources Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences, 2MOE Key Laboratory of Major Diseases in Children, National Key Discipline of Pediatrics, National Clinical Research Center for Respiratory Diseases, Beijing Key Laboratory of Pediatric Respiratory Infection Diseases, Beijing Pediatric Research Institute, Beijing Children's Hospital, Capital Medical University, 3State Key Laboratory of Molecular Developmental Biology, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences A simple method for obtaining NK and T cell clones from CAEBV patients was developed with high efficiency, a small amount of peripheral blood, and a low-dose of IL-2. Neuroscience An Improved Method for Collection of Cerebrospinal Fluid from Anesthetized Mice Nastasia K-H Lim1,2, Visse Moestrup3,4, Xiao Zhang1, Wen-An Wang5, Arne Møller3,4,6, Fu-De Huang1,4 1Shanghai Advanced Research Institute, University of Chinese Academy of Sciences, Chinese Academy of Science, 2Shanghai Institute of Materia Medica, University of Chinese Academy of Sciences, Chinese Academy of Science, 3Center of Functionally Integrative Neuroscience, Department of Clinical Medicine, Aarhus University, 4Sino-Danish Center for Education and Research (SDC), 5Department of Neurology, Xinhua Hospital Chongming Branch Affiliated to Shanghai Jiao Tong University School of Medicine, 6Department of Nuclear Medicine and PET-centre, Aarhus University Hospital This protocol describes an improved technique for the abundant collection of cerebrospinal fluid (CSF) with no contamination from blood. With greater sample collection and purity, more analyses can be performed using CSF to further our understanding of diseases that affect the brain and spinal cord. Genetics CRISPR/Cas9-mediated Targeted Integration In Vivo Using a Homology-mediated End Joining-based Strategy Xuan Yao1,2, Xing Wang1,2, Junlai Liu2,3,4, Linyu Shi1, Pengyu Huang3, Hui Yang1 1Institute of Neuroscience, State Key Laboratory of Neuroscience, Key Laboratory of Primate Neurobiology, CAS Center for Excellence in Brain Science and Intelligence Technology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 2College of Life Sciences, University of Chinese Academy of Sciences, 3School of Life Science and Technology, Shanghai Tech University, 4Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences The clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system provides a promising tool for genetic engineering, and opens up the possibility of targeted integration of transgenes. We describe a homology-mediated end joining (HMEJ)-based strategy for efficient DNA targeted integration in vivo and targeted gene therapies using CRISPR/Cas9. Developmental Biology Application of Aorta-gonad-mesonephros Explant Culture System in Developmental Hematopoiesis Junhua Lv1,2, Feng Liu1,2 1State Key Laboratory of Membrane Biology, Institute of Zoology, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences This protocol describes using cultured Aorta-Gonad-Mesonephros for expression analyses, colony-forming units in the culture and spleen, and long-term reconstitution to determine the effect of regulatory factors and signaling pathways on hematopoietic stem cell development. This has been demonstrated as an effective system for studying hematopoietic stem cell biology and function. Engineering The Modular Design and Production of an Intelligent Robot Based on a Closed-Loop Control Strategy Libo Zhang1, Junjie Zhu2, Hao Ren2, Dongdong Liu3, Dan Meng4, Yanjun Wu1, Tiejian Luo5 1Institute of Software, Chinese Academy of Sciences, 2Hunan University, 3Fuyang Normal University, 4Anhui Normal University, 5University of Chinese Academy of Sciences We present a protocol on modular design and production of intelligent robots to help scientific and technical workers design intelligent robots with special production tasks based on personal needs and individualized design. Neuroscience Live-cell Measurement of Odorant Receptor Activation Using a Real-time cAMP Assay Yuetian Zhang*1, Yi Pan*1, Hiroaki Matsunami2,3, Hanyi Zhuang1,4 1Department of Pathophysiology, Key Laboratory of Cell Differentiation and Apoptosis of National Ministry of Education, Shanghai Jiao Tong University School of Medicine, 2Department of Molecular Genetics and Microbiology, Duke University Medical Center, 3Department of Neurobiology, Duke Institute for Brain Sciences, Duke University Medical Center, 4Institute of Health Science, Chinese Academy of Science/Shanghai Jiao Tong University School of Medicine Characterizing the function of odorant receptors serves an indispensable part in the deorphanization process. We describe a method to measure the activation of odorant receptors in real time using a cAMP assay. Behavior An Automated Rapid Iterative Negative Geotaxis Assay for Analyzing Adult Climbing Behavior in a Drosophila Model of Neurodegeneration Wenze Cao*1,2,3, Li Song*1,2,3, Jingjing Cheng1,2,3, Na Yi1,2,3, Luyi Cai1,2,3, Fu-de Huang4,5, Margaret Ho1,2,3 1Research Center for Translational Medicine, Tongji University School of Medicine, 2Key Laboratory of Arrhythmias of the Ministry of Education of China, Shanghai East Hospital, Tongji University School of Medicine, 3Department of Anatomy and Neurobiology, Tongji University School of Medicine, 4Shanghai Advanced Research Institute, University of Chinese Academy of Sciences, Chinese Academy of Sciences, 5Sino-Danish College, University of Chinese Academy of Sciences, Chinese Academy of Sciences This step-by-step protocol analyzes Drosophila negative geotaxis behavior using an automated multi-cylinder system that hosts hundreds of flies and synchronizes their action by an electric motor. Upon synchronization, fly negative geotaxis behavior is assayed, digitally recorded, and analyzed using the self-designed RflyDetection software. Developmental Biology A Novel Mammary Fat Pad Transplantation Technique to Visualize the Vessel Generation of Vascular Endothelial Stem Cells Qing Cissy Yu1, Wenqian Song1, Dengwen Lai1, Yi Arial Zeng1 1State Key Laboratory of Cell Biology, CAS Center for Excellence in Molecular Cell Science, Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences This work demonstrates a novel approach to assess the proliferation, differentiation, and vessel-forming potential of vascular endothelial stem cells (VESCs) through mammary fat pad transplantation followed by whole-mount tissue preparation for microscopic observation. A lineage tracing strategy to investigate the behavior of VESCs in vivo is also presented. Developmental Biology Whole-cell Patch-clamp Recordings of Isolated Primary Epithelial Cells from the Epididymis Bao Li Zhang1,2,3, Da Yuan Gao1,2,3, Xiao Xu Zhang1,3, Shuo Shi4, Winnie Shum1 1School of Life Science and Technology, ShanghaiTech University, 2Institute of Biochemistry and Cell Biology, Shanghai Institutes for Biological Sciences, Chinese Academy of Sciences, 3University of Chinese Academy of Sciences, 4Shanghai Institute for Advanced Immunochemical Studies, ShanghaiTech University We present a protocol that combines cell isolation and whole-cell patch-clamp recording to measure the electrical properties of the primary dissociated epithelial cells from the rat cauda epididymides. This protocol allows for investigation of the functional properties of primary epididymal epithelial cells to further elucidate the physiological role of the epididymis. Developmental Biology Differentiating Chondrocytes from Peripheral Blood-derived Human Induced Pluripotent Stem Cells Yueying Li2, Yong Hai1, Jiayu Chen3, Tie Liu1 1Department of Orthopedics, Beijing Chao-Yang Hospital, Capital Medical University, 2Key Laboratory of Genomic and Precision Medicine, Beijing Institute of Genomics, Chinese Academy of Sciences, 3Clinical and Translational Research Center of Shanghai First Maternity & Infant Hospital, School of Life Sciences and Technology, Tongji University We present a protocol to generate a chondrogenic lineage from human peripheral blood (PB) via induced pluripotent stem cells (iPSCs) using an integration-free method, which includes embryoid body (EB) formation, fibroblastic cells expansion, and chondrogenic induction. Bioengineering Production of Chemicals by Klebsiella pneumoniae Using Bamboo Hydrolysate as Feedstock Dong Wei1,2, Jinjie Gu1, Zhongxi Zhang1, Chenhong Wang1, Dexin Wang1, Chul Ho Kim3, Biao Jiang1, Jiping Shi1,4, Jian Hao1 1Lab of Biorefinery, Shanghai Advanced Research Institute, Chinese Academy of Sciences, 2University of Chinese Academy of Sciences, 3Biorefinery Research Center, Jeonbuk Branch Institute, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 4School of Life Science and Technology, ShanghaiTech University Bamboo powder was pretreated with NaOH and enzymatically hydrolyzed. The hydrolysate of bamboo was used as the feedstock for 2,3-butanediol, R-acetoin, 2-ketogluconic acid, and xylonic acid production by Klebsiella pneumoniae. Immunology and Infection An All-on-chip Method for Rapid Neutrophil Chemotaxis Analysis Directly from a Drop of Blood Ke Yang*1,2,3, Jiandong Wu*3,4, Ling Zhu1, Yong Liu1, Michael Zhang5, Francis Lin3,4,6,7 1Institute of Applied Technology, Hefei Institutes of Physical Science, Chinese Academy of Sciences, 2University of Science and Technology of China, 3Department of Physics and Astronomy, University of Manitoba, 4Department of Biosystems Engineering, University of Manitoba, 5Seven Oaks General Hospital, 6Department of Immunology, University of Manitoba, 7Department of Biological Sciences, University of Manitoba This article provides the detailed method of performing a rapid neutrophil chemotaxis assay by integrating the on-chip neutrophil isolation from whole blood and the chemotaxis test on a single microfluidic chip. Genetics Protocols for C-Brick DNA Standard Assembly Using Cpf1 Shi-Yuan Li1, Guo-Ping Zhao1, Jin Wang1 1Institute of Plant Physiology and Ecology, Shanghai Insititutes for Biological Sciences, Chinese Academy of Sciences CRISPR-associated protein Cpf1 can be guided by a specially designed CRISPR RNA (crRNA) to cleave double-stranded DNA at desired sites, generating sticky ends. Based on this characteristic, a DNA assembly standard (C-Brick) was established, and a protocol detailing its use is described here. Medicine Optimized Analysis of In Vivo and In Vitro Hepatic Steatosis Aoyuan Cui1, Zhimin Hu1, Yamei Han1, Yi Yang2, Yu Li1 1CAS Key Laboratory of Nutrition and Metabolism, Institute for Nutritional Sciences, Chinese Academy of Sciences, 2Suzhou Niumag Analytical Instrument Corporation Here, optimized methods to generate in vivo and in vitro models of hepatic steatosis and to analyze the steatotic phenotypes and related physiological parameters are described. Immunology and Infection A Simple Red Blood Cell Lysis Method for the Establishment of B Lymphoblastoid Cell Lines Xi Liu1,2, Chongfeng Xu2, Ziyuan Duan2 1Wuhan Institute of Virology, Chinese Academy of Sciences, 2Genetic Resources Center, Institute of Genetics and Developmental Biology, Chinese Academy of Sciences A simple red blood cell lysis method for establishing B-LCLs was developed with high immortalization efficiency, requiring a small amount of blood and saving time from initiation to cryopreservation. Bioengineering Fluorescence Biomembrane Force Probe: Concurrent Quantitation of Receptor-ligand Kinetics and Binding-induced Intracellular Signaling on a Single Cell Yunfeng Chen*1, Baoyu Liu*2, Lining Ju*3, Jinsung Hong*1, Qinghua Ji4,5, Wei Chen6, Cheng Zhu1 1Woodruff School of Mechanical Engineering, Petit Institute for Bioengineering and Biosciences, Georgia Institute of Technology, 2Coulter Department of Biomedical Engineering, Georgia Institute of Technology, 3Charles Perkins Centre, The University of Sydney, 4Institute of Biophysics, Laboratory of RNA Biology, Chinese Academy of Sciences, 5University of Chinese Academy of Sciences, 6School of Medicine and Collaborative Innovation Center for Diagnosis and Treatment of Infectious Diseases, Zhejiang University We describe a technique for concurrently measuring force-regulated single receptor-ligand binding kinetics and real-time imaging of calcium signaling in a single T lymphocyte. Biology Measuring Spatial and Temporal Ca2+ Signals in Arabidopsis Plants Xiaohong Zhu1, Aaron Taylor2, Shenyu Zhang1, Dayong Zhang1,3, Ying Feng1,4, Gaimei Liang1,5, Jian-Kang Zhu1,6 1Department of Horticulture and Landscape Architecture, Purdue University, 2Bindley Bioscience Center, Purdue University, 3Institute of Biotechnology, Jiangsu Academy of Agricultural Sciences, 4College of Environmental & Resource Science, Zhejiang University, 5Dryland Agriculture Research Centre, Shanxi Academy of Agricultural Sciences, 6Shanghai Center for Plant Stress Biology, Chinese Academy of Sciences Ca2+ signaling regulates diverse biological processes in plants. Here we present approaches for monitoring abiotic stress induced spatial and temporal Ca2+ signals in Arabidopsis cells and tissues using the genetically encoded Ca2+ indicators Aequorin or Case12. Biology Single-molecule Imaging of Gene Regulation In vivo Using Cotranslational Activation by Cleavage (CoTrAC) Zach Hensel1, Xiaona Fang1,2,3, Jie Xiao1 1Department of Biophysics and Biophysical Chemistry, Johns Hopkins University School of Medicine, 2Changchun Institute of Applied Chemistry, Chinese Academy of Sciences, 3Department of Physics, Jilin University We describe a fluorescence microscopy method, Co-Translational Activation by Cleavage (CoTrAC), to image the production of protein molecules in live cells with single-molecule precision without perturbing the protein's functionality. This method has been used to follow the stochastic expression dynamics of a transcription factor, the λ repressor CI 1. Biology Collection, Isolation and Enrichment of Naturally Occurring Magnetotactic Bacteria from the Environment Zachery Oestreicher1, Steven K. Lower1,2, Wei Lin3, Brian H. Lower2 1School of Earth Sciences, The Ohio State University, 2School of Environment & Natural Resources, The Ohio State University, 3Institute of Geology and Geophysics, Chinese Academy of Sciences We demonstrate a method to collect magnetotactic bacteria (MTB) that can be applied to natural waters. MTB can be isolated and enriched from sediment samples using a relatively simple setup that takes advantage of the bacteria's natural magnetism. Isolated MTB can then be examined in detail using both light and electron microscopy.