Zhejiang University

19 articles published in JoVE

• Medicine

  
  Laminectomy for the Removal of Thoracic Ossification of the Ligamentum Flavum (TOLF) Using Ultrasonic and Conventional Osteotomes Junhui Liu^1,2, Zhi Shan^1,2, Yilei Chen^1,2, Fengdong Zhao^{1,2 1}Department of Orthopedics, Sir Run Run Shaw Hospital, School of Medicine, Zhejiang University, ²Key Laboratory of Musculoskeletal System Degeneration and Regeneration Translational Research of Zhejiang Province We present a methodology for using an ultrasonic osteotome combined with a conventional osteotome for laminectomy in thoracic ossification of the ligamentum flavum. This is a relatively safe and easy-to-learn method that avoids the perioperative complications associated with the conventional method.

• Biology

  
  Analyzing Starvation-Induced Autophagy in the Drosophila melanogaster Larval Fat Body Kexin Shi¹, Chao Tong^{1 1}Ministry of Education Key Laboratory of Biosystems Homeostasis and Protection and Innovation Center for Cell Signaling Network, Life Sciences Institute, Zhejiang University The present protocol describes the induction of autophagy in the Drosophila melanogaster larval fat body via nutrient depletion and analyzes changes in autophagy using transgenic fly strains.

• Biology

  
  Profiling of H3K4me3 Modification in Plants using Cleavage under Targets and Tagmentation Xiaoyuan Tao*^1,2, Mengtao Gao*³, Siyuan Wang*¹, Xueying Guan^{1,4 1}College of Agriculture and Biotechnology, Zhejiang University, ²Central Laboratory, State Key Laboratory for Managing Biotic and Chemical Threats to the Quality and Safety of Agro-products, Zhejiang Academy of Agricultural Sciences, ³State Key Laboratory of Crop Genetics and Germplasm Enhancement, Cotton Hybrid R & D Engineering Center (the Ministry of Education), College of Agriculture, Nanjing Agricultural University, ⁴Hainan Institute of Zhejiang University Cleavage under targets and tagmentation (CUT&Tag) is an efficient chromatin epigenomic profiling strategy. This protocol presents a refined CUT&Tag strategy for the profiling of histone modifications in plants.

• Biology

  
  High-Throughput Metabolic Profiling for Model Refinements of Microalgae Amnah Alzahmi^1,2, Sarah Daakour¹, Diana Charles El Assal³, Bushra S. Dohai³, Amphun Chaiboonchoe³, Weiqi Fu^3,4, David R. Nelson¹, Alexandra Mystikou¹, Kourosh Salehi-Ashtiani^{1,3 1}Center for Genomics and Systems Biology (CGSB), New York University Abu Dhabi Research Institute, ²Department of Biology, United Arab Emirates University, ³Laboratory of Algal, Systems, and Synthetic Biology (LASSB), Division of Science and Math, New York University Abu Dhabi, ⁴Department of Marine Science, Ocean College, Zhejiang University This protocol demonstrates the use of a phenotype microarray (PM) technology platform to define metabolic requirements of Chlamydomonas reinhardtii, a green microalga, and refine an existing metabolic network model.

• Environment

  
  Isolation of Quartz Grains for Optically Stimulated Luminescence (OSL) Dating of Quaternary Sediments for Paleoenvironmental Research Liliana C. Marin¹, Steven L. Forman¹, Victoria T. Todd¹, Connor Mayhack¹, Ashley Gonzalez¹, Peng Liang^{2 1}Geoluminescence Dating Research Laboratory, Dept. of Geosciences, Baylor University, ²School of Earth Sciences, Zhejiang University This protocol is for the isolation of quartz grains by size for luminescence dating of sediments. Outlined are physical cleansing and chemical digestions by soaking sequentially in H2O2, HCl, HF, and HCl again to isolate quartz grains. The quartz purity is quantified with microscopic assessment, Raman spectroscopy, and IR depletion ratio.

• Cancer Research

  
  Establishing a Competing Risk Regression Nomogram Model for Survival Data Lunpo Wu^1,2, Chenyang Ge³, Hongjuan Zheng⁴, Haiping Lin⁵, Wei Fu⁶, Jianfei Fu^{4 1}Department of Gastroenterology, Second Affiliated Hospital of Zhejiang University School of Medicine, ²Institute of Gastroenterology, Zhejiang University, ³Department of Colorectal Surgery, Afiliated Jinhua Hospital, Zhejiang University School of Medicine, ⁴Department of Medical Oncology, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, ⁵Department of Hepatobiliary Pancreatic Surgery, Affiliated Jinhua Hospital, Zhejiang University School of Medicine, ⁶Division of Oncology, Johns Hopkins University School of Medicine Presented here is a protocol to build nomograms based on the Cox proportional hazards regression model and competing risk regression model. The competing method is a more rational method to apply when competing events are present in the survival analysis.

• Immunology and Infection

  
  Localization and Quantification of Begomoviruses in Whitefly Tissues by Immunofluorescence and Quantitative PCR Fei-Xue Ban¹, Tian-Yan Yin¹, Qi Guo¹, Li-Long Pan¹, Yin-Quan Liu¹, Xiao-Wei Wang^{1 1}Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogen and Insects, Institute of Insect Sciences, Zhejiang University We describe an immunofluorescence and quantitative PCR method for the localization and quantification of begomoviruses in insect tissues. The immunofluorescence protocol can be used to colocalize viral and vector proteins. The quantitative PCR protocol can be extended to quantify viruses in whole whitefly bodies and virus-infected plants.

• Bioengineering

  
  Protocols of 3D Bioprinting of Gelatin Methacryloyl Hydrogel Based Bioinks Mingjun Xie^1,2, Kang Yu^1,2, Yuan Sun^1,2, Lei Shao^1,2, Jing Nie^1,2, Qing Gao^1,2, Jingjiang Qiu³, Jianzhong Fu^1,2, Zichen Chen^1,2, Yong He^{1,2 1}State Key Laboratory of Fluid Power and Mechatronic Systems, School of Mechanical Engineering, Zhejiang University, ²Key Laboratory of 3D Printing Process and Equipment of Zhejiang Province, School of Mechanical Engineering, Zhejiang University, ³School of Mechanics and Safety Engineering, Zhengzhou University Presented here is a method for the 3D bioprinting of gelatin methacryloyl.

• Neuroscience

  
  The Detection of 5-Hydroxymethylcytosine in Neural Stem Cells and Brains of Mice Yingliang Zhuang^1,2, Junchen Chen^1,2, Weize Xu¹, Qiang Shu¹, Xuekun Li^{1,2 1}The Children's Hospital, School of Medicine, Zhejiang University, ²The Institute of Translational Medicine, School of Medicine, Zhejiang University Here, we present a protocol to detect 5-hydroxymethylcytosine in cells and brain tissues, utilizing immunofluorescence staining and DNA dot-blot methods.

• Neuroscience

  
  Investigating Alterations in Caecum Microbiota After Traumatic Brain Injury in Mice Liang Wen*¹, Wendong You*¹, Yadong Wang*¹, Yuanrun Zhu¹, Hao Wang¹, Xiaofeng Yang^{1 1}Emergency and Trauma Center, The International Medical Center, The First Affiliated Hospital, School of Medicine, Zhejiang University Presented here is a protocol to induce diffuse traumatic brain injury using a lateral fluid percussion device followed by the collection of the caecum content for gut microbiome analysis.

• Genetics

  
  Identifying Mutations by High Resolution Melting in a TILLING Population of Rice Shan Li¹, Ying-Ping Yu¹, Song-Mei Liu¹, Hao-Wei Fu², Jian-Zhong Huang³, Qing-Yao Shu¹, Yuan-Yuan Tan^{4 1}National Key Laboratory of Rice Biology, Zhejiang Provincial Key Laboratory of Crop Genetic Resources, Institute of Crop Science, Zhejiang University, ²Jiaxing Academy of Agricultural Sciences, ³Institute of Nuclear Agricultural Sciences, Zhejiang University, ⁴The New Countryside Development Institute at Zhejiang University In this article, we present the protocol that is described as high-resolution melting analysis (HRM)-based Target Induced Local Lesions In Genomes (TILLING). This method utilizes fluorescence changes during the melting of the DNA duplex and is suitable for high-throughput screening of both insertion/deletion (Indel) and single base substition (SBS).

• Environment

  
  Visualization of Flow Field Around a Vibrating Pipeline Within an Equilibrium Scour Hole Dawei Guan^1,2, Yee-Meng Chiew², Maoxing Wei³, Shih-Chun Hsieh^{4 1}Key Laboratory of Coastal Disaster and Defense, Ministry of Education, Hohai University, ²School of Civil and Environmental Engineering, Nanyang Technological University, ³Ocean College, Zhejiang University, ⁴AXESEA Information Technology Co. LTD The goal of the protocol is to enable visualization of the detailed flow fields and determination of the near-boundary shear and normal stresses within an equilibrium scour hole induced by a vibrating pipeline.

• Behavior

  
  Design and Use of an Apparatus for Presenting Graspable Objects in 3D Workspace Kedi Xu^1,2,3, Junjun Chen^1,2, Guanghao Sun^1,2, Yaoyao Hao^1,3, Shaomin Zhang^1,2,3, Xingchen Ran^1,2, Weidong Chen^1,2, Xiaoxiang Zheng^{1,2,3 1}Qiushi Academy for Advanced Studies, Zhejiang University, ²Department of Biomedical Engineering, Zhejiang University, ³Key Laboratory of Biomedical Engineering of Ministry of Education, Zhejiang University Presented here is a protocol to build an automatic apparatus that guides a monkey to perform the flexible reach-to-grasp task. The apparatus combines a 3D translational device and turning table to present multiple objects in an arbitrary position in 3D space.

• Immunology and Infection

  
  Effects of Taste Signaling Protein Abolishment on Gut Inflammation in an Inflammatory Bowel Disease Mouse Model Ya-Wen Du*¹, Qun Liu*¹, Xiao-Cui Luo¹, Dong-Xiao Zhao¹, Jian-Bo Xue¹, Pu Feng², Robert F. Margolskee², Hong Wang², Liquan Huang^{1,2 1}College of Life Sciences, Zhejiang University, ²Monell Chemical Senses Center Here we present a protocol to investigate the effect of the nullification of gustation-related genes on immune responses in a dextran sulfate sodium (DSS)-induced inflammatory bowel disease (IBD) mouse model.

• Neuroscience

  
  Experimental Models to Study the Neuroprotection of Acidic Postconditioning Against Cerebral Ischemia Yanrong Zheng*¹, Zhe Shen*¹, Xiaoli Wu¹, Lei Jiang¹, Weiwei Hu¹, Zhong Chen¹, Xiangnan Zhang^{1 1}Institute of Pharmacology & Toxicology, College of Pharmaceutical Sciences, Department of Pharmacology, Key Laboratory of Medical Neurobiology of The Ministry of Health of China, Zhejiang Province Key Laboratory of Neurobiology, Zhejiang University Acidic postconditioning protects against cerebral ischemia. Here we present two models to execute APC. They are achieved respectively by transferring corticostriatal slices to acidic buffer after oxygen-glucose deprivation in vitro and by inhaling 20% CO2 after middle cerebral artery occlusion in vivo.

• Biology

  
  Methods for the Extraction of Endosymbionts from the Whitefly Bemisia tabaci Dan-Tong Zhu¹, Xin-Ru Wang¹, Fei-Xue Ban¹, Chi Zou¹, Shu-Sheng Liu¹, Xiao-Wei Wang^{1 1}Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogen and Insects, Institute of Insect Sciences, Zhejiang University Here, we present a protocol to isolate endosymbionts from the whitefly Bemisia tabaci through dissection and filtration. After amplification, the DNA samples are suitable for subsequent sequencing and study of the mutualism between endosymbionts and whitefly.

• Biology

  
  Detection of Ligand-activated G Protein-coupled Receptor Internalization by Confocal Microscopy Jingwen Yang¹, Yunjun Yan¹, Xiaowei Xiang¹, Yuchao Xu¹, Naiming Zhou², Tianming Wang^{1 1}National Engineering Research Center of Marine Facilities Aquaculture, Marine Science College, Zhejiang Ocean University, ²Institute of Biochemistry, College of Life Sciences, Zhejiang University This protocol describes confocal microscopy detection of G protein-coupled receptor (GPCR) internalization in mammalian cells. It includes the basic cell culture, transfection, and confocal microscopy procedure and provides an efficient and easily interpretable method to detect the subcellular localization and internalization of fusion-expressed GPCR.

• 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 Ji^4,5, Wei Chen⁶, Cheng Zhu^{1 1}Woodruff 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 Zhang^1,3, Ying Feng^1,4, Gaimei Liang^1,5, Jian-Kang Zhu^{1,6 1}Department 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.