JoVE Logo

Sign In

University of Guelph

34 ARTICLES PUBLISHED IN JoVE

image

Biology

Adenovirus-mediated Genetic Removal of Signaling Molecules in Cultured Primary Mouse Embryonic Fibroblasts
Steve P. Hawley *1, Melanie K. B. Wills *1, Nina Jones 1
1Department of Molecular and Cellular Biology, University of Guelph

In this video we use an adenovirus carrying the Cre recombinase gene to infect primary mouse embryonic fibroblasts carrying a floxed Rac1 allele.

image

Neuroscience

Monitoring Cleaved Caspase-3 Activity and Apoptosis of Immortalized Oligodendroglial Cells using Live-cell Imaging and Cleaveable Fluorogenic-dye Substrates Following Potassium-induced Membrane Depolarization
Graham S.T. Smith 1, Janine A.M. Voyer-Grant 1, George Harauz 1
1Department of Molecular and Cellular Biology, University of Guelph

Live-cell imaging of caspase-3 mediated apoptosis in immortalized N19-oligodendrocyte cell cultures using the NucView 488 caspase-3 substrate. This technique is applicable for programmed cell death assays in real-time in a variety of cell types and tissues.

image

Behavior

A Novel Procedure for Evaluating the Reinforcing Properties of Tastants in Laboratory Rats: Operant Intraoral Self-administration
AnneMarie Levy 1, Cheryl L. Limebeer 1, Justin Ferdinand 1, Ucal Shillingford 1, Linda A. Parker 1, Francesco Leri 1
1Department of Psychology, University of Guelph

The current study evaluates a novel procedure for assessing the reinforcing effects of palatable solutions in laboratory rats: intraoral self-administration. To this end, operant responding (i.e. lever pressing) for intraoral infusions of sweet solutions at different concentrations was measured on continuous and progressive ratios schedules of reinforcement.

image

Chemistry

Towards Biomimicking Wood: Fabricated Free-standing Films of Nanocellulose, Lignin, and a Synthetic Polycation
Karthik Pillai 1,2,3, Fernando Navarro Arzate 2,4, Wei Zhang 2,5, Scott Renneckar 1,2,5,6
1Institute for Critical Technology and Applied Science, Virginia Tech, 2Macromolecules and Interfaces Institute, Virginia Tech, 3Institute for Food Safety and Health, Illinois Institute of Technology- Moffett Campus, 4Wood, Cellulose, and Paper Research Department, University of Guadalajara, 5Department of Sustainable Biomaterials, Virginia Tech, 6Sustainable Nanotechnology Interdisciplinary Graduate Education Program, Virginia Tech

The objective of this research was to form synthetic plant cell wall tissue using layer-by-layer assembly of nanocellulose fibrils and isolated lignin assembled from dilute aqueous suspensions.  Surface measurement techniques of quartz crystal microbalance and atomic force microscopy were used to monitor the formation of the polymer-polymer nanocomposite material.

image

Medicine

Vascular Balloon Injury and Intraluminal Administration in Rat Carotid Artery
Wei Zhang 1, Mohamed Trebak 1
1Nanobioscience, State University of New York College of Nanoscale Science and Engineering (SUNY CNSE)

This protocol uses a balloon catheter to cause an intraluminal injury on the rat carotid artery and henceforth elicit neointimal hyperplasia. This is a well-established model for studying the mechanisms of vascular remodeling in response to injury. It is also widely used to determine the validity of potential therapeutic approaches.

image

Bioengineering

Surface Potential Measurement of Bacteria Using Kelvin Probe Force Microscopy
Eric Birkenhauer 1, Suresh Neethirajan 1
1BioNano Laboratory, School of Engineering, University of Guelph

Here, we present a protocol explaining the use of Kelvin probe force microscopy as a tool for generating high resolution nano-scale surface potential maps. This tool was applied to assess the role of surface potential on the binding capacity of microorganisms to substrate surfaces.

image

Bioengineering

Methods for Characterizing the Co-development of Biofilm and Habitat Heterogeneity
Xiaobao Li 1, Jisun L. Song 2, Alessandro Culotti 1, Wei Zhang 1, David L. Chopp 3, Nanxi Lu 1, Aaron I. Packman 1
1Department of Civil and Environmental Engineering, Northwestern University, 2Department of Chemical and Biological Engineeering, Northwestern University, 3Department of Applied Mathematics and Engineering Sciences, Northwestern University

Biofilms have complex interactions with their surrounding environment. To comprehensively investigate biofilm-environment interactions, we present here a series of methods to create heterogeneous chemical environment for biofilm development, to quantify local flow velocity, and to analyze mass transport in and around biofilm colonies.

image

Biochemistry

Sheathless Capillary Electrophoresis–Mass Spectrometry for Metabolic Profiling of Biological Samples
Wei Zhang 1, M. Can Gulersonmez 1, Thomas Hankemeier 1, Rawi Ramautar 1
1Division of Analytical Biosciences, Leiden Academic Center for Drug Research, Leiden University

A protocol for metabolic profiling of biological samples by capillary electrophoresis–mass spectrometry using a sheathless porous tip interface design is presented.

image

Engineering

Two-way Valorization of Blast Furnace Slag: Synthesis of Precipitated Calcium Carbonate and Zeolitic Heavy Metal Adsorbent
Evangelos Georgakopoulos 1, Rafael M. Santos 2, Yi Wai Chiang 3, Vasilije Manovic 4
1Department of Offshore, Process and Energy Engineering, Cranfield University, 2School of Applied Chemical and Environmental Sciences, Sheridan College Institute of Technology and Advanced Learning, 3School of Engineering, University of Guelph, 4Carbon Systems Engineering, Centre for Combustion, Carbon Capture and Storage, Cranfield University

A protocol for the parallel production of precipitated calcium carbonate and zeolitic material from blast furnace slag via mineral carbonation and alkaline hydrothermal conversion, respectively, is presented. The performance of the zeolitic material towards nickel adsorption is tested.

image

Genetics

Analysis of Cap-binding Proteins in Human Cells Exposed to Physiological Oxygen Conditions
Sara Timpano *1, Gaelan Melanson *1, Sonia L. Evagelou 1, Brianna D. Guild 1, Erin J. Specker 1, James Uniacke 1
1Department of Molecular and Cellular Biology, University of Guelph

Here, we present human cell culture protocols to analyze translation initiation factors that bind the 5' cap of mRNA during physiological oxygen conditions. This method utilizes an Agarose-linked m7GTP cap analog and is suitable to investigate cap-binding factors and their interacting partners.

image

Developmental Biology

Dissection of Larval Zebrafish Gonadal Tissue
Xinjian Wang *1, Sijie Chen *1, Wei Zhang 1, Yiyuan Ren 1, Quan Zhang 1, Gang Peng 1
1Institutes of Brain Science, State Key Laboratory of Medical Neurobiology and Collaborative Innovation Center for Brain Science, Fudan University

Here, we present a protocol for isolating gonadal tissue of larval zebrafish, which will facilitate investigations of zebrafish sex differentiation and maintenance.

image

Neuroscience

Imaging Neurons within Thick Brain Sections Using the Golgi-Cox Method
Emma L. Louth 1, Charles D. Sutton 1, Ari L. Mendell 1, Neil J. MacLusky 1, Craig D.C. Bailey 1
1Department of Biomedical Sciences, Ontario Veterinary College, University of Guelph

We present a protocol for using the Golgi-Cox staining method in thick brain sections, in order to visualize neurons with long dendritic trees contained within single tissue samples. Two variants of this protocol are also presented that involve cresyl violet counterstaining, and the freezing of unprocessed brains for long-term storage.

image

Biology

Detecting the Lyme Disease Spirochete, Borrelia Burgdorferi, in Ticks Using Nested PCR
Melanie K. B. Wills 1, Andrea M. Kirby 1, Vett K. Lloyd 1
1Department of Biology, Mount Allison University

Nested PCR is a sensitive, specific, and straightforward technique that can be applied to tick DNA extracts to probe for Borrelia burgdorferi, the causative agent of Lyme disease. The initial PCR experiment uses gene-specific primers to generate long amplicons, which then become templates for a subsequent reaction using internal primers.

image

Engineering

Novel Photoacoustic Microscopy and Optical Coherence Tomography Dual-modality Chorioretinal Imaging in Living Rabbit Eyes
Chao Tian 1, Wei Zhang 2,3, Van Phuc Nguyen 1, Xueding Wang 2,4, Yannis M. Paulus 1,2
1Kellogg Eye Center, Department of Ophthalmology and Visual Sciences, University of Michigan, 2Department of Biomedical Engineering, University of Michigan, 3Institute of Biomedical Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, 4Department of Radiology, University of Michigan

This manuscript describes the novel setup and operating procedure of a photoacoustic microscopy and optical coherence tomography dual-modality system for noninvasive, label-free chorioretinal imaging of larger animals, such as rabbits.

image

Neuroscience

A Rat Model of Central Fatigue Using a Modified Multiple Platform Method
Weiyue Zhang *1, Wei Zhang *1, Ning Dai *1, Chenxia Han 2, Fengzhi Wu 1, Xu Wang 1, Libo Tan 1, Jie Li 1, Feng Li 1, Qingjia Ren 3
1School of Traditional Chinese Medicine, Beijing University of Chinese Medicine, 2Department of Integrated Traditional Chinese and Western Medicine, West China Hospital, Sichuan University, 3Institute of Tibetan Medicine, Tibetan Traditional Medical College

Here, we present a protocol to introduce a rat model of central fatigue using the modified multiple platform method (MMPM).

image

Medicine

Introduction of Intracapsular Rotary-cut Procedures (IRCP): A Modified Hysteromyomectomy Procedures Facilitating Fertility Preservation
Liping Zeng 1,2, Weixia Wei 1,2, Huiru Tang 1,2, Fang He 1,2, Shilin Zhong 1,2, Wei Zhang 1,2, Yuting Xiang 1, Xinfeng Qu 3, Ruifang Wu 1,2
1Department of Obstetrics and Gynecology, Peking University Shenzhen Hospital, China, 2Shenzhen Key Laboratory of Gynecological Diagnostic Technology Research, China, 3Specialist of SZSM Project, Peking University Shenzhen Hospital, China

Here, we present a protocol for performing an intracapsular rotary-cut procedure (IRCP), a modified laparoscopic intracapsular myomectomy that promotes fertility preservation.

image

Chemistry

Methionine Functionalized Biocompatible Block Copolymers for Targeted Plasmid DNA Delivery
Yang Wu *1, Wei Zhang *2, Jian Zhang 2, Zhi-Xiang Mao 3, Li Ding 2, Hao Li 3, Rong Ma 1, Jin-Hai Tang 2
1Research Center of Clinical Oncology, Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & Nanjing Medical University Affiliated Cancer Hospital, 2Department of General Surgery, The First Affiliated Hospital with Nanjing Medical University, 3School of Clinical Medicine, Xuzhou Medical University

This work presents the preparation of methionine functionalized biocompatible block copolymers (mBG) via the reversible addition-fragmentation chain transfer (RAFT) method. The plasmid DNA complexing ability of the obtained mBG and their transfection efficiency were also investigated. The RAFT method is very beneficial for polymerizing monomers containing special functional groups.

image

Biology

Bovine Mammary Gland Biopsy Techniques
Veridiana L. Daley 1,4, Charles Dye 2, Sophie H. Bogers 3, R. Michael Akers 4, Francisco C. Rodriguez 3, John P. Cant 5, John Doelman 5, Peter Yoder 4, Karan Kumar 2, Dane Webster 6, Mark D. Hanigan 4
1National Animal Nutrition Program, a National Research Support Project (NRSP-9), Department of Animal and Food Sciences, University of Kentucky, 2School of Performing Arts, Virginia Tech, 3Department of Large Animal Clinical Sciences, The Virginia-Maryland College of Veterinary Medicine, Virginia Tech, 4Department of Dairy Science, Virginia Tech, 5Department of Animal Biosciences, University of Guelph, 6School of Visual Arts, Virginia Tech

This article presents a bovine mammary gland biopsy using core and needle biopsy tools. Harvested tissue can be used for cell culture or to assess mammary physiology and metabolism including gene expression, protein expression, protein modifications, immunohistochemistry, and metabolite concentrations.

image

Bioengineering

Protein Kinase C-delta Inhibitor Peptide Formulation using Gold Nanoparticles
Hisato Konoeda 1, Hong Yang 2, Chengliang Yang 1, Annette Gower 1, Chun Xu 1, Wei Zhang 1, Mingyao Liu 1,3
1Latner Thoracic Surgery Research Laboratories, Toronto General Hospital Research Institute, University Health Network, 2Respiratory Medicine Research Laboratory, Institute of Translation Medicine, Shanghai General Hospital, Shanghai Jiaotong University, 3Institute of Medical Science, Faculty of Medicine, University of Toronto

We have previously used a gold nanoparticle peptide hybrid to intravenously deliver a synthetic peptide, protein kinase C-delta inhibitor, which reduced ischemia-reperfusion-induced acute lung injury. Here we show the detailed protocol of the drug formulation. Other intracellular peptides can be formulated similarly.

image

Medicine

A Training and Testing System for Performing Vascular Reconstruction In Vitro
Yue Wang 1,2, Lijun Mu 3, Wei Zhang 1,2, Huan Chen 1,2, Qingshan Li 1,2, Aihua Shi 1, Bo Tang 2, Xiaogang Zhang 1,2, Dinghui Dong 1,2, Yi Lv 1,2
1National Local Joint Engineering Research Center for Precision Surgery and Regenerative Medicine, First Affiliated Hospital of Xi'an Jiaotong University, 2Department of Hepatobiliary Surgery, First Affiliated Hospital of Xi'an Jiaotong University, 3Department of General Surgery, First Affiliated Hospital of Xi'an Jiaotong University

Here we present a training and testing system where a trainee can complete manual vascular reconstruction in vitro individually using a magnetic anchoring technique. The system can also be used to test the quality of reconstruction.

image

Immunology and Infection

Antibiotic Dereplication Using the Antibiotic Resistance Platform
Haley L. Zubyk 1, Georgina Cox 2, Gerard D. Wright 1
1Department of Biochemistry and Biomedical Sciences, M.G. DeGroote Institute for Infectious Disease Research, McMaster University, 2Department of Molecular and Cellular Biology, University of Guelph

We describe a platform that utilizes a library of isogenic antibiotic resistant Escherichia coli for the dereplication of antibiotics. The identity of an antibiotic produced by bacteria or fungi can be deduced by the growth of E. coli expressing its respective resistance gene. This platform is economically effective and time-efficient.

image

Biology

Field Identification of Matricaria chamomilla using a Portable qPCR System
Zhengxiu Yang 1, Zheng Quan 2, Tiffany Chua 3, Leo Li 1, Yanjun Zhang 3, Silva Babajanian 2, Francesco Buongiorno 4, Isabella Della Noce 4, Lorenzo Colombo 4, Steven Newmaster 5, Tricia Chua 3, Peter Chang 3, Gary Swanson 3, Zhengfei Lu 2
1Herbalife NatSource (Hunan) Natural Products Co., Ltd., 2Corporate Quality Laboratory, Herbalife International of America, Inc., 3Corporate Quality, Herbalife International of America, Inc., 4Hyris Ltd, 5Natural Health Product Research Alliance, University of Guelph

Presented here is a protocol for field identification of Matricaria chamomilla using a portable qPCR system. This easy-to-perform protocol is ideal as a method to confirm the identity of a botanical species at locations where access to laboratory equipment and expertise is limited, such as farms and warehouses.

image

Chemistry

Capillary Electrophoresis Mass Spectrometry Approaches for Characterization of the Protein and Metabolite Corona Acquired by Nanomaterials
Andrew J. Chetwynd *1, Wei Zhang *2, Klaus Faserl *3, James A. Thorn 4, Iseult Lynch 1, Rawi Ramautar 2, Herbert H. Lindner 3
1School of Geography Earth and Environmental Sciences, University of Birmingham, 2Biomedical Microscale Analytics, Leiden University, 3Institute of Medical Biochemistry, Medical University of Innsbruck, 4AB Sciex UK Ltd

Here we present a protocol to characterize the complete biomolecular corona, proteins, and metabolites, acquired by nanomaterials from biofluids using a capillary electrophoresis – mass spectrometry approach.

image

Developmental Biology

A Protocol for Immunohistochemistry and RNA In-situ Distribution within Early Drosophila Embryo
Wei Zhang *1, Xinjuan Lei *1, Xin Zhou *2,3, Boling He 1, Liqin Xiao 1, Huimin Yue 1, Shulin Wang 1, Yuting Sun 1, Yajun Wu 1, Liyang Wang 1,4, George Ghartey-Kwansah 1, Odell D. Jones 5, Joseph L. Bryant 6, MengMeng Xu 7, Jianjie Ma 3, Xuehon Xu 1
1National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China/CGDB, Shaanxi Normal University College of Life Sciences, 2Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Shaanxi Key Laboratory of Brain disorders, Institute of Basic & Translational Medicine, Xi'an Medical University, 3Ohio State University College of Medicine, 4Hematology-Oncology Division, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 5University of Pennsylvania ULAR, 6University of Maryland School of Medicine, 7Columbia University Medical Center

Here, we describe a protocol for detection and localization of Drosophila embryo protein and RNA from collection to pre-embedding and embedding, immunostaining, and mRNA in situ hybridization.

image

Biochemistry

Semi-Quantitative Analysis of Peptidoglycan by Liquid Chromatography Mass Spectrometry and Bioinformatics
Erin M. Anderson 1, Neil A. Greenwood 1, Dyanne Brewer 2, Cezar M. Khursigara 1
1Department of Molecular and Cellular Biology, University of Guelph, 2Mass Spectrometry Facility, University of Guelph

This protocol covers a detailed analysis of peptidoglycan composition using liquid chromatography mass spectrometry coupled with advanced feature extraction and bioinformatic analysis software.

image

Immunology and Infection

Label-Free Quantitative Proteomics Workflow for Discovery-Driven Host-Pathogen Interactions
Brianna Ball 1, Arjun Sukumaran 1, Jennifer Geddes-McAlister 1
1Molecular and Cellular Biology Department, University of Guelph

Here, we present a protocol to profile the interplay between host and pathogen during infection by mass spectrometry-based proteomics. This protocol uses label-free quantification to measure changes in protein abundance of both host (e.g., macrophages) and pathogen (e.g., Cryptococcus neoformans) in a single experiment.

image

Environment

Monitoring Pedogenic Inorganic Carbon Accumulation Due to Weathering of Amended Silicate Minerals in Agricultural Soils.
Reza Khalidy *1, Fatima Haque *1, Yi Wai Chiang 1, Rafael M. Santos 1
1School of Engineering, University of Guelph

The verification method described here is adaptable for monitoring pedogenic inorganic carbon sequestration in various agricultural soils amended with alkaline earth metal silicate-containing rocks, such as wollastonite, basalt, and olivine. This type of validation is essential for carbon credit programs, which can benefit farmers that sequester carbon in their fields.

image

Neuroscience

Acquisition of Resting-State Functional Magnetic Resonance Imaging Data in the Rat
Diana J. Wallin *1,2, Emily D. K. Sullivan *1,2, Elise M. Bragg 1, Jibran Y. Khokhar 2,4, Hanbing Lu 2,3, Wilder T. Doucette 1,2
1Dartmouth-Hitchcock Medical Center, 2Geisel School of Medicine at Dartmouth, 3National Institute on Drug Abuse, National Institutes of Health, 4University of Guelph

This protocol describes a method for obtaining stable resting-state functional magnetic resonance imaging (rs-fMRI) data from a rat using low dose isoflurane in combination with low dose dexmedetomidine.

image

Immunology and Infection

Production of Adeno-Associated Virus Vectors in Cell Stacks for Preclinical Studies in Large Animal Models
Amira D. Rghei 1, Brenna A. Y. Stevens *1, Sylvia P. Thomas *1, Jacob G. E. Yates *1, Benjamin M. McLeod 1, Khalil Karimi 1, Leonardo Susta 1, Byram W. Bridle 1, Sarah K. Wootton 1
1Department of Pathobiology, University of Guelph

Here we provide a detailed procedure for large-scale production of research-grade AAV vectors using adherent HEK 293 cells grown in cell stacks and affinity chromatography purification. This protocol consistently yields >1 x 1013 vector genomes/mL, providing vector quantities appropriate for large animal studies.

image

Biochemistry

Using Phage Display to Develop Ubiquitin Variant Modulators for E3 Ligases
Olivia Roscow 1, Wei Zhang 1
1Department of Molecular and Cellular Biology, College of Biological Science, University of Guelph

Ubiquitination is a critical protein post-translational modification, dysregulation of which has been implicated in numerous human diseases. This protocol details how phage display can be utilized to isolate novel ubiquitin variants that can bind and modulate the activity of E3 ligases that control the specificity, efficiency, and patterns of ubiquitination.

image

Behavior

Assessment of Mouse Judgment Bias through an Olfactory Digging Task
Aileen MacLellan *1, Agustina Resasco *2,3, Lauren Young 4, Georgia Mason 1
1Department of Integrative Biology, University of Guelph, 2Institute of Cell Biology and Neurosciences, National Scientific and Technical Research Council-University of Buenos Aires, 3Laboratory of Experimental Animals, Faculty of Veterinary Sciences, National University of La Plata, 4Department of Animal Biosciences, University of Guelph

This article provides a detailed description of a novel mouse judgment bias protocol. Evidence of this olfactory digging task's sensitivity to affective state is also demonstrated and its utility across diverse research fields is discussed.

image

Cancer Research

Production of High-Titer Recombinant Newcastle Disease Virus from Allantoic Fluid
Jacob G. E. Yates 1, Alexander Leacy 1, Phuc H. Pham 1, Nicole Zielinska 1, Emily A. Tusnadi 1, Leonardo Susta 1, Sarah K. Wootton 1
1Department of Pathobiology, University of Guelph

Here we provide a detailed procedure for production, purification, and quantification of high-titer recombinant Newcastle disease virus. This protocol consistently yields > 6 × 109 plaque-forming units/mL, providing virus quantities appropriate for in vivo animal studies. Additional quality control assays to ensure safety in vivo are described.

image

Biology

Assessing the Putative Anticryptococcal Properties of Crude and Clarified Extracts from Mollusks
Davier Gutierrez-Gongora 1, Fouad Raouf-Alkadhimi 1, Ryan S. Prosser 2, Jennifer Geddes-McAlister 1
1Molecular and Cellular Biology Department, University of Guelph, 2School of Environmental Sciences, University of Guelph

The human fungal pathogen Cryptococcus neoformans produces a variety of virulence factors (e.g., peptidases) to promote its survival within the host. Environmental niches represent a promising source of novel natural peptidase inhibitors. This protocol outlines the preparation of extracts from mollusks and the assessment of their effect on fungal virulence factor production.

image

Innovative Animal Models Of Cardiac Remodeling: Development And Evaluation
Feng Hua Yang 1, W. Glen Pyle 2,3, Yihua Bei 4
1Cardiovascular Model Research Center, Guangdong Laboratory Animals Monitoring Institute, 2IMPART Investigator Team, Dalhousie Medicine, 3Department of Biomedical Sciences, University of Guelph, 4School of Medicine, Shanghai University

Innovative Animal Models Of Cardiac Remodeling: Development And Evaluation

JoVE Logo

Privacy

Terms of Use

Policies

Research

Education

ABOUT JoVE

Copyright © 2024 MyJoVE Corporation. All rights reserved