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33 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Isolation and Expansion of the Adult Mouse Neural Stem Cells Using the Neurosphere Assay
Hassan Azari 1,2, Maryam Rahman 2, Sharareh Sharififar 2, Brent A. Reynolds 2
1 Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran , 2Department of Neurosurgery, University of Florida

This video protocol demonstrates the neurosphere assay method to generate and expand neural stem cells from the adult mouse periventricular region, and provides technical insights to ensure one can achieve reproducible neurosphere cultures.

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Neuroscience

Establishing Embryonic Mouse Neural Stem Cell Culture Using the Neurosphere Assay
Hassan Azari 1,2, Sharareh Sharififar 2, Maryam Rahman 2, Saeed Ansari 2, Brent A. Reynolds 2
1Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran , 2Department of Neurosurgery, The University of Florida

This video protocol demonstrates the application of the neurosphere assay for the isolation and expansion of neural stem cells from the ganglionic eminences of embryonic day 14-mouse brain.

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Neuroscience

Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells
Hassan Azari 1,2, Sharon A. Louis 3, Sharareh Sharififar 1, Vinata Vedam-Mai 1, Brent A. Reynolds 1
1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, 3STEMCELL Technologies, Inc.

This video protocol demonstrates how to discriminate and enumerate bona fide neural stem cells in a mixed population of neural precursor cells using the neural colony-forming cell assay.

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Medicine

Intraluminal Middle Cerebral Artery Occlusion (MCAO) Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Mice
Saeed Ansari *1, Hassan Azari *1,2, Douglas J. McConnell 1, Aqeela Afzal 1, J Mocco 1
1Department of Neurosurgery, University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences

The intraluminal middle cerebral artery occlusion (MCAO) model is the most frequent used model among experimental ischemic stroke models. Here we will demonstrate the entire model in detail with the guide of Laser Doppler flowmetry, and its representative results.

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Biology

Pouring and Running a Protein Gel by reusing Commercial Cassettes
Alexander C. Hwang 1, Paris H. Grey 1, Katrina Cuddy 1, David G. Oppenheimer 1,2,3
1Department of Biology, University of Florida , 2UF Genetics Institute, University of Florida , 3Plant Molecular & Cellular Biology Program, University of Florida

Our protocol demonstrates how to pour multiple protein gels at a time by recycling Invitrogen Nupage Novex minigel cassettes, and inexpensive materials purchased at a home improvement store. This economical and streamlined method includes a way to store the gels at 4°C for a few weeks. By re-using the plastic gel cassettes from commercially available gels, labs that run frequent protein gels can save significant costs and help the environment.

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Biology

Detection of Protein Interactions in Plant using a Gateway Compatible Bimolecular Fluorescence Complementation (BiFC) System
Gang Tian 1, Qing Lu 2, Li Zhang 2, Susanne E. Kohalmi 1, Yuhai Cui 2
1Department of Biology, University of Western Ontario, 2Southern Crop Protection and Food Research Centre, Agriculture and Agri-Food Canada

We have developed a technique to test protein-protein interactions in plant. A yellow fluorescent protein (YFP) is split into two non-overlapping fragments. Each fragment is cloned in-frame to a gene of interest via Gateway system, enabling expression of fusion proteins. Reconstitution of YFP signal only occurs when the inquest proteins interact.

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Neuroscience

Isolation and Expansion of Human Glioblastoma Multiforme Tumor Cells Using the Neurosphere Assay
Hassan Azari 1,2, Sebastien Millette 1, Saeed Ansari 1, Maryam Rahman 1, Loic P. Deleyrolle 1, Brent A. Reynolds 1
1Department of Neurosurgery, University of Florida , 2Department of Anatomical Sciences, Shiraz University of Medical Sciences

This video protocol demonstrates the isolation and expansion of stem like cells from surgically resected human glioblastoma mutliforme (GBM) tumor tissue using the neurosphere assay culture method.

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Neuroscience

The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytometry
Hassan Azari 1,2, Sharareh Sharififar 1, Jeff M. Fortin 1, Brent A. Reynolds 1
1Department of Neurosurgery, The University of Florida, 2Laboratory for Stem Cell Research, Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

This video protocol demonstrates a novel method for the generation and subsequent purification of neuronal progenitor cells from a renewable source of neural stem cells (NSCs) based on their physical (size and internal granularity) and fluorescent properties using flow cytometry technology.

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Biology

Profiling Thiol Redox Proteome Using Isotope Tagging Mass Spectrometry
Jennifer Parker 1, Ning Zhu 2, Mengmeng Zhu 2, Sixue Chen 1,2,3,4
1Plant Molecular and Cellular Biology Program, University of Florida , 2Department of Biology, University of Florida , 3Interdisciplinary Center for Biotechnology Research, University of Florida , 4Genetics Institute, University of Florida

Reactive oxygen species level is elevated when cells encounter stress conditions. Here we show the example of 3'-3' diaminobenzidine staining as well as cysTMT labeling and mass spectrometry to profile the redox proteome in Pseudomonas syringae treated tomato leaves.

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Biology

GC-based Detection of Aldononitrile Acetate Derivatized Glucosamine and Muramic Acid for Microbial Residue Determination in Soil
Chao Liang 1,2, Harry W. Read 2, Teri C. Balser 2,3
1DOE-Great Lakes Bioenergy Research Center, University of Wisconsin, Madison, 2Department of Soil Science, University of Wisconsin, Madison, 3Department of Soil and Water Science, University of Florida

We describe a method protocol for the GC-based analysis of the aldonitrile acetate derivatives of glucosamine and muramic acid extracted from soil. For elucidation of the chemical mechanism, we also present a strategy to confirm the structure of the derivative and the ion fragments formed upon electron ionization.

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Neuroscience

Single Cell Measurement of Dopamine Release with Simultaneous Voltage-clamp and Amperometry
Kaustuv Saha 1, Jarod Swant 1, Habibeh Khoshbouei 1,2
1Department of Neuroscience, University of Florida , 2Department of Psychiatry, University of Florida

The amperometric technique measures dopamine release from a single cell by detecting the oxidative current produced by spontaneous dopamine oxidization. Simultaneous voltage clamp and amperometry methodology reveal the mechanistic relationship between the overall "activity" of dopamine transporter and the regulatory role of this activity on the reverse transport of dopamine.

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Medicine

Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE)
Loic P. Deleyrolle 1, Mark R. Rohaus 1, Jeff M. Fortin 1, Brent A. Reynolds 1, Hassan Azari 1,2
1Department of Neurosurgery, The University of Florida, 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

This video protocol demonstrates the application of the fluorescent dye carboxyfluorescein succinimidyl ester (CFSE) for the identification and separation of different sub-populations of cells in human glioblastoma based on frequency of cell division.

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Medicine

Collection Protocol for Human Pancreas
Martha L. Campbell-Thompson 1, Emily L. Montgomery 1, Robin M. Foss 1, Kerwin M. Kolheffer 1, Gerald Phipps 1, Lynda Schneider 1, Mark A. Atkinson 1
1Department of Pathology, Immunology, and Laboratory Medicine, University of Florida

This video demonstrates a dissection procedure for processing human pancreas into multiple storage formats. Anatomical orientation is maintained throughout the pancreatic regions to allow definition of regional islet composition and density.

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Medicine

Staining Protocols for Human Pancreatic Islets
Martha L. Campbell-Thompson 1, Tiffany Heiple 1, Emily Montgomery 1, Li Zhang 1, Lynda Schneider 1
1Department of Pathology, Immunology, and Laboratory Medicine, University of Florida

This video demonstrates procedures for characterization of human pancreatic islets using hematoxylin and eosin (H&E) and immunohistochemistry (IHC). Pancreatic sections from head, body, and tail regions are stained by both H&E and IHC to determine islet endocrine composition (insulin, glucagon, and pancreatic polypeptide), cell replication (Ki67), and inflammatory infiltrates (H&E, CD3). The uncinate region is localized using IHC for pancreatic polypeptide.

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Biology

Transmitting Plant Viruses Using Whiteflies
Jane E. Polston 1, H. Capobianco 1
1Deptartment of Plant Pathology, University of Florida

Improving our understanding of and our ability to manage many of the insect-transmitted plant viruses requires the use of the vector. Insect transmission of plant viruses is a tritrophic interaction, and as such requires the manipulation of insects, virus, and plant. Vectors must be reared in large numbers and manipulated in such a way as to insure high rates of transmission to test plants. The basics of rearing and manipulating the whitefly, Bemisa tabaci (Hemiptera: Aleyrodidae), a vector of plant viruses belonging to several emerging plant virus genera, representing a large number of economically significant viruses is presented.

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Medicine

Endothelin-1 Induced Middle Cerebral Artery Occlusion Model for Ischemic Stroke with Laser Doppler Flowmetry Guidance in Rat
Saeed Ansari 1, Hassan Azari 2, Kenneth J. Caldwell 1, Robert W. Regenhardt 3, Vishnumurthy S. Hedna 4, Micheal F. Waters 4, Brian L. Hoh 1, Adam P. Mecca 3
1Department of Neurosurgery, University of Florida , 2Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran , 3Department of Physiology and Functional Genomics, University of Florida , 4Department of Neurology, University of Florida

Several animal models of cerebral ischemia have been developed to simulate the human condition of stroke. This protocol describes the endothelin-1 (ET-1) induced middle cerebral artery occlusion (MCAO) model for ischemic stroke in rats. In addition, important considerations, advantages, and shortcomings of this model are discussed.

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Medicine

Utilizing a Cranial Window to Visualize the Middle Cerebral Artery During Endothelin-1 Induced Middle Cerebral Artery Occlusion
Robert W. Regenhardt 1, Saeed Ansari 2, Hassan Azari 3, Kenneth J. Caldwell 2, Adam P. Mecca 1
1Department of Physiology and Functional Genomics, University of Florida , 2Department of Neurosurgery, McKnight Brain Institute, University of Florida , 3Department of Anatomical Sciences, Shiraz University of Medical Sciences, Shiraz, Iran

This article describes a method for visualizing rat cerebral arteries through a cranial window using temporal craniectomy in order to view proximal portions of the middle cerebral artery (Figure 1). This versatile method can be combined with various techniques of drug delivery to measure cerebral artery reactivity in vivo.

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JoVE Journal

Using High Resolution Computed Tomography to Visualize the Three Dimensional Structure and Function of Plant Vasculature
Andrew J. McElrone 1,2, Brendan Choat 3, Dilworth Y. Parkinson 4, Alastair A. MacDowell 4, Craig R. Brodersen 5
1U.S. Department of Agriculture, 2Department of Viticulture and Enology, University of California - Davis, 3Hawkesbury Institute for the Environment, University of Western Sydney, 4Advanced Light Source, Lawrence Berkeley National Lab, 5Citrus Research & Education Center, University of Florida

High resolution x-ray computed tomography (HRCT) is a non-destructive diagnostic imaging technique that can be used to study the structure and function of plant vasculature in 3D. We demonstrate how HRCT facilitates exploration of xylem networks across a wide range of plant tissues and species.

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Neuroscience

Vertical T-maze Choice Assay for Arthropod Response to Odorants
Lukasz Stelinski 1, Siddharth Tiwari 1
1Entomology and Nematology Department, University of Florida

A vertical, T-maze olfactometer is described for assaying the behavioral response of arthropods. The olfactometer allows the experimenter to measure choices performed by test subjects when subjected to two potential odor fields. Both attraction to and repulsion from odorants can be measured with this device.

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Biology

Purification of Transcripts and Metabolites from Drosophila Heads
Kurt Jensen 1, Jonatan Sanchez-Garcia 1, Caroline Williams 2, Swati Khare 1, Krishanu Mathur 1, Rita M. Graze 3, Daniel A. Hahn 2, Lauren M. McIntyre 3, Diego E. Rincon-Limas 1,4, Pedro Fernandez-Funez 1,4
1Department of Neurology, McKnight Brain Institute, University of Florida , 2Department of Entomology and Nematology, University of Florida , 3Genetics Institute, Department of Molecular Genetics and Microbiology, University of Florida , 4McKnight Brain Institute, Department of Neuroscience, Genetics Institute, Center for Translational Research on Neurodegenerative Diseases, and Center for Movement Disorders and Neurorestoration, University of Florida

We describe here the procedures for the extraction and purification of mRNA and metabolites from Drosophila heads. We are applying these techniques to better understand the cellular perturbations underlying neuronal degeneration. These methodologies can be easily scaled and adapted for other "omic" projects.

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Bioengineering

Protocol for Relative Hydrodynamic Assessment of Tri-leaflet Polymer Valves
Sharan Ramaswamy 1, Manuel Salinas 1, Rob Carrol 2, Karla Landaburo 1, Xavier Ryans 1, Cynthia Crespo 1, Ailyn Rivero 1, Faris Al-Mousily 3,4, Curt DeGroff 3, Mark Bleiweis 3, Hitomi Yamaguchi 2
1Tissue Engineered Mechanics, Imaging and Materials Laboratory, Department of Biomedical Engineering, Florida International University, 2Department of Mechanical and Aerospace Engineering, University of Florida , 3College of Medicine, University of Florida , 4King Faisal Specialty Hospital and Research Center, Jeddah, Saudi Arabia

There has been renewed interest in developing polymer valves. Here, the objectives are to demonstrate the feasibility of modifying a commercial pulse duplicator to accommodate tri-leaflet geometries and to define a protocol to present polymer valve hydrodynamic data in comparison to native and prosthetic valve data collected under near-identical conditions.

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Behavior

Use of the Operant Orofacial Pain Assessment Device (OPAD) to Measure Changes in Nociceptive Behavior
Ethan M. Anderson 1,2, Richard Mills 3, Todd A. Nolan 4, Alan C. Jenkins 4, Golam Mustafa 1, Chris Lloyd 3, Robert M. Caudle 1,2, John K. Neubert 4
1Department of Oral and Maxillofacial Surgery, University of Florida College of Dentistry, 2Department of Neuroscience, McKnight Brain Institute, University of Florida College of Medicine , 3Stoelting Co., 4Department of Orthodontics, University of Florida

We present a user-friendly, high-throughput operant system for the evaluation of pain behaviors in awake, conscious rodents. The Orofacial Pain Assessment Device (OPAD) can assess pain through a reward/conflict paradigm thus providing a more humane way of testing. This protocol will yield more clinically relevant and translational data from rodents.

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Biology

Measurement of Heme Synthesis Levels in Mammalian Cells
Jagmohan Hooda 1, Maksudul Alam 1, Li Zhang 1
1Department of Molecular and Cell Biology, Center for Systems Biology, University of Texas at Dallas

Altered intracellular heme levels are associated with common diseases such as cancer. Thus, there is a need to measure heme biosynthesis levels in diverse cells. The goal of this protocol is to provide a fast and sensitive method to measure and compare the levels of heme synthesis in different cells.

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Biology

Stomata Tape-Peel: An Improved Method for Guard Cell Sample Preparation
Sheldon Lawrence II 1,2,3, Quiying Pang 2,3,4, Wenwen Kong 2,3, Sixue Chen 1,2,3,5
1Plant Molecular and Cellular Biology Program, University of Florida, 2Department of Biology, University of Florida, 3Genetics Institute (UFGI), University of Florida, 4Alkali Soil Natural Environmental Science Center, Northeast Forestry University, 5Interdisciplinary Center for Biotechnology Research (ICBR), University of Florida

This protocol describes a method of preparing enriched stomatal guard cells that is useful for physiological and other biological studies.

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Immunology and Infection

High-Efficiency Generation of Antigen-Specific Primary Mouse Cytotoxic T Cells for Functional Testing in an Autoimmune Diabetes Model
Howard W. Davidson 1, Joseph Ray Cepeda 2, Nitin S. Sekhar 2, Junying Han 2, Ling Gao 3, Tomasz Sosinowski 1, Li Zhang 2
1Barbara Davis Center for Childhood Diabetes, University of Colorado Denver, 2Department of Medicine, Endocrinology, Diabetes & Metabolism, Baylor College of Medicine, 3Scientific Center, Shandong Provincial Hospital affiliated to Shandong University

This article describes a protocol for the generation of antigen-specific CD8 T cells, and their expansion in vitro, with the aim of yielding high numbers of functional T cells for use in vitro and in vivo.

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JoVE Core

International Expert Consensus and Recommendations for Neonatal Pneumothorax Ultrasound Diagnosis and Ultrasound-guided Thoracentesis Procedure
Jing Liu 1,2, Dalibor Kurepa 3, Francesco Feletti 4,5, Almudena Alonso-Ojembarrena 6, Jovan Lovrenski 7, Roberto Copetti 8, Erich Sorantin 9, Javier Rodriguez-Fanjul 10, Karishma Katti 3, Andrea Aliverti 4, Huayan Zhang 11,12, Misun Hwang 13, Tsu F. Yeh 14, Cai-Bao Hu 15, Xing Feng 16, Ru-Xin Qiu 1,2, Jing-Han Chi 17, Li-Li Shang 18, Guo-Rong Lyu 19, Shao-Zheng He 20, Yan-Fen Chai 21, Zhan-Jun Qiu 22, Hai-Ying Cao 2,23, Yue-Qiao Gao 1,2, Xiao-Ling Ren 1,2, Guo Guo 1,24, Li Zhang 1,2, Ying Liu 1,2, Wei Fu 1,2, Zu-Lin Lu 1,2, Hong-Lei Li 1,2
1Department of Neonatology and NICU, Beijing Chaoyang District Maternal and Child Healthcare Hospital, 2The National Neonatal Lung Ultrasound Training Base, 3Division of Neonatal-Perinatal Medicine, Cohen Children's Medical Center, 4Department of Electronics, Information and Bioengineering, Politecnico di Milano, 5Dipartimento di Diagnostica per Immagini, Ausl della Romagna, S. Maria delle Croci Hospital, 6Neonatal Intensive Care Unit, Puerta del Mar University Hospital, 7Faculty of Medicine, University of Novi Sad, Serbia, Institute for Children and Adolescents Health Care of Vojvodina, 8Emergency Department, University Hospital of Cattinara, 9Division of Pediatric Radiology, Department of Radiology, Medical University Graz, 10Pediatric Intensive Care Unit, Pediatric Service Hospital Joan XXIII Tarragona, University Rovira i Virgil, 11Center for Newborn Care, Guangzhou Women and Children's Medical Center, 12Division of Neonatology, Children's Hospital of Philadelphia, 13Section of Neonatal Imaging, Department of Radiology, Children's Hospital of Philadelphia, 14Maternal Child Health Research institute, Taipei Medical University and China Medical University, 15Intensive Care Unit, Zhejiang Hospital, 16Department of Neonatology, Children's Hospital of Soochow University, 17Department of Neonatology and NICU, Bayi Children's Hospital Affiliated to the Seventh Medical Center of Chinese PLA General Hospital, 18Intensive Care Unit, Second Affiliated Hospital of Heilongjiang University of Chinese Medicine, 19Collaborative Innovation Center for Maternal and Infant Health Service Application Technology, Quanzhou Medical College, 20Department of Ultrasound, Second Affiliated Hospital of Fujian Medical University, 21Department of Emergency Medicine, Tianjin Medical University General Hospital, 22Department of Emergency and Critical Care Medicine, Affiliated Hospital of Traditional Chinese Medicine, 23Department of Ultrasound, GE Healthcare, 24The Neonatal Intensive Care Unit, Fifth Medical Center of Chinese PLA General Hospital

Pneumothorax is a common emergency and critical disease in newborn infants that needs rapid, clear diagnosis and timely treatment. Diagnosis and treatment based on chest X-rays are associated with delayed management and radiation damage. Lung ultrasound (US) provides useful guidance for rapid, accurate diagnosis and the precise thoracentesis of pneumothorax.

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Immunology and Infection

Observing Islet Function and Islet-Immune Cell Interactions in Live Pancreatic Tissue Slices
Mollie K. Huber 1, Denise M. Drotar 2,3,4, Helmut Hiller 1, Maria L. Beery 1, Paul Joseph 1, Irina Kusmartseva 1, Stephan Speier 2,3,4, Mark A. Atkinson 1, Clayton E. Mathews 1, Edward A. Phelps 5
1Department of Pathology, Immunology, and Laboratory Medicine, University of Florida, 2Paul Langerhans Institute Dresden (PLID) of the Helmholtz Zentrum München at the University Clinic Carl Gustav Carus of Technische Universität Dresden, 3Institute of Physiology, Faculty of Medicine, Technische Universität Dresden, 4German Center for Diabetes Research (DZD), 5J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida

This study presents the application of live pancreatic tissue slices to the study of islet physiology and islet-immune cell interactions.

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Biology

Preparing Irradiated and Marked Male Aedes aegypti Mosquitoes for Release in an Operational Sterile Insect Technique Program
Bianca J. Moreno 1, Robert L. Aldridge 1, Seth C. Britch 1, Barbara E. Bayer 1, Jedidiah Kline 1, Daniel A. Hahn 2, Chao Chen 2, Kenneth J. Linthicum 1
1US Department of Agriculture, Agricultural Research Service Center for Medical, Agricultural, & Veterinary Entomology, 2Department of Entomology and Nematology, University of Florida

The sterile insect technique (SIT) is used to control specific, medically important mosquito populations that may be resistant to chemical controls. Here, we describe a method of mass rearing and preparation of sterile male mosquitoes for release in an operational SIT program targeting the Aedes aegypti mosquito.

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Biology

Isolation of Primary Rat Hepatocytes with Multiparameter Perfusion Control
Inn Chuan Ng *1, Li Zhang *2, Narelle Nichola Yi Ying Shen 3, Yun Ting Soong 4, Chan Way Ng 5, Phoebe Kang Sheing Koh 1, Yan Zhou 3, Hanry Yu 1,3,4,5,6
1Department of Physiology & The Institute for Digital Medicine (WisDM), National University of Singapore, 2College of Agriculture and Biology, Zhongkai University of Agriculture and Engineering, 3Mechanobiology Institute, National University of Singapore, 4Institute of Bioengineering and Nanotechnology, A*STAR, 5NUS Graduate School for Integrative Sciences and Engineering, 6CAMP, Singapore-MIT Alliance for Research and Technology

This protocol details the use of a special intravenous catheter, standardized sterile disposable tubing, temperature control complemented by real-time monitoring, and an alarm system for two-step collagenase perfusion procedure to improve the consistency in the viability, yield, and functionality of isolated primary rat hepatocytes.

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JoVE Journal

Genome-wide Analysis of Histone Modifications Distribution using the Chromatin Immunoprecipitation Sequencing Method in Magnaporthe oryzae
Zechi Wu *1, Wanyu Sun *1, Sida Zhou 1, Li Zhang 1, Xinyu Zhao 1, Yang Xu 1, Weixiang Wang 1
1Beijing Key Laboratory of New Technology in Agricultural Application, National Demonstration Center for Experimental Plant Production Education, Department of Agronomy, Beijing University of Agriculture

Here, we present a protocol to analyze the genome-wide distribution of histone modifications, which can identify new target genes in the pathogenesis of M. oryzae and other filamentous fungi.

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Measuring Cerebral Vasospasm after Subarachnoid Hemorrhage in Mice
William S. Dodd 1, Brandon Lucke-Wold 1, Koji Hosaka 1, Brian L. Hoh 1
1Department of Neurosurgery, College of Medicine, University of Florida

The present protocol provides a detailed procedure for inducing subarachnoid hemorrhage in mice via autologous blood injection to the anterior circulation and measuring delayed cerebral vasospasm by vascular gel casting.

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Biology

Combining 3D Magnetic Force Actuator and Multi-Functional Fluorescence Imaging to Study Nucleus Mechanobiology
Miao Huang 1,5, Heyang Wang 1,5, Alfredo A. Delgado 1, Tyler A. Reid 1, Julian Long 2, Shu Wang 3,5, Hayley Sussman 4, Juan Guan 5,6,7, Hitomi Yamaguchi 1, Xin Tang 1,5,8,9
1Department of Mechanical and Aerospace Engineering, Herbert Wertheim College of Engineering, University of Florida, 2Department of Materials Science and Engineering, University of Florida, 3Department of Biostatistics, University of Florida, 4Department of Biomedical Engineering, College of Engineering (COE), University of Delaware (UD), 5UF Health Cancer Center, University of Florida, 6Department of Physics, College of Liberal Arts and Sciences, University of Florida, 7Department of Anatomy and Cell Biology, College of Medicine, University of Florida, 8J. Crayton Pruitt Family Department of Biomedical Engineering, University of Florida, 9Department of Physiology and Functional Genomics, University of Florida

This study presents a new protocol to directly apply mechanical force on the cell nucleus through magnetic microbeads delivered into the cytoplasm and to conduct simultaneous live-cell fluorescent imaging.

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Behavior

Assessment of Nerve Injury-Induced Mechanical Hypersensitivity in Rats Using an Orofacial Operant Pain Assay
Basak Donertas-Ayaz 1, Ariana C. Brice-Tutt 2, Wendi L. Malphurs 2, Darrice Montgomery 2, Richard H. Mills 3, John K. Neubert 2, Robert M. Caudle 1
1Department of Oral and Maxillofacial Surgery, University of Florida, Gainesville, 2Department of Orthodontics, University of Florida, Gainesville, 3Stoelting Co. Wood Dale

This protocol describes the assessment of mechanical hypersensitivity in a rat model of neuropathic orofacial pain using an operant-based orofacial pain assessment device.

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