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Ohio State University

15 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Stereotaxic Surgery for Excitotoxic Lesion of Specific Brain Areas in the Adult Rat
Elizabeth D. Kirby 1, Kelly Jensen 2, Ki A. Goosens 3, Daniela Kaufer 1,4
1Helen Wills Neuroscience Institute, University of California Berkeley, 2Office of Laboratory Animal Care, University of California Berkeley, 3McGovern Institute for Brain Research & The Department of Brain and Cognitive Science, Massachusetts Institute of Technology, 4Integrative Biology Department, University of California Berkeley

Targeted ablation of specific brain region(s) by infusion of an excitotoxin using stereotaxic coordinates is described. This technique could also be adapted for infusion of other chemicals into the rat brain.

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JoVE Journal

Bead Aggregation Assays for the Characterization of Putative Cell Adhesion Molecules
Michelle R. Emond 1, James D. Jontes 1
1Department of Neuroscience, Ohio State University

Here we present a simple, rapid method for characterizing the intrinsic adhesive properties of putative cell adhesion molecules. The secreted, epitope-tagged ectodomain of a cell adhesion molecule is captured from the culture medium on small, uniform functionalized beads. These beads can then be used immediately in simple bead aggregation assays.

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Biology

Intravenous Injections in Neonatal Mice
Sara E. Gombash Lampe 1, Brian K. Kaspar 2, Kevin D. Foust 1
1Department of Neuroscience, Ohio State University, 2Center for Gene Therapy, Nationwide Children's Hospital Research Institute, Ohio State University

Animal models of pediatric disease can experience early onset and aggressive disease progression. Clinically relevant therapy delivery to young mouse models can be technically difficult. This protocol describes a non-invasive intravenous injection method for newborn mice within the first two postnatal days of life.

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Medicine

Method of Isolated Ex Vivo Lung Perfusion in a Rat Model: Lessons Learned from Developing a Rat EVLP Program
Kevin Nelson 1,2, Christopher Bobba 1,2, Emre Eren 3, Tyler Spata 4, Malak Tadres 2, Don Hayes, Jr. 5,6, Sylvester M. Black 3,7, Samir Ghadiali *1,2,3, Bryan A. Whitson *2,3,4
1Department of Biomedical Engineering, Ohio State University Wexner Medical Center, 2Davis Heart & Lung Research Institute, Ohio State University Wexner Medical Center, 3The Collaboration for Organ Perfusion, Protection, Engineering and Regeneration (COPPER) Laboratory, Ohio State University Wexner Medical Center, 4Division of Cardiac Surgery, Department of Surgery, Ohio State University Wexner Medical Center, 5Departments of Pediatrics and Internal Medicine, Ohio State University, 6Advanced Lung Disease Program, Lung and Heart-Lung Transplant Programs, Nationwide Children's Hospital, 7Division of Transplantation, Department of Surgery, Ohio State University Wexner Medical Center

Ex-Vivo Lung Perfusion (EVLP) has allowed lung transplantation in humans to become more readily available by enabling the ability to assess organs and expand the donor pool. Here, we describe the development of a rat EVLP program and refinements that allow for a reproducible model for future expansion.

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Engineering

Electron Channeling Contrast Imaging for Rapid III-V Heteroepitaxial Characterization
Julia I. Deitz 1, Santino D. Carnevale 2, Steven A. Ringel 3, David W. McComb 3, Tyler J. Grassman 1,2
1Department of Materials Science and Engineering, The Ohio State University, 2Department of Electrical and Computer Engineering, The Ohio State University, 3Institute of Materials Research, The Ohio State University

The use of electron channeling contrast imaging in a scanning electron microscope to characterize defects in III-V/Si heteroexpitaxial thin films is described. This method yields similar results to plan-view transmission electron microscopy, but in significantly less time due to lack of required sample preparation.

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Biochemistry

Isolation of Cognate RNA-protein Complexes from Cells Using Oligonucleotide-directed Elution
Gatikrushna Singh *1, Sarah M. Fritz *2, Arnaz Ranji 2, Deepali Singh 3, Kathleen Boris-Lawrie 1,2
1Department of Veterinary & Biomedical Sciences, University of Minnesota, 2Department of Veterinary Biosciences, Ohio State University, 3School of Biotechnology, Gautam Buddha University

This manuscript describes an approach to isolate select cognate RNPs formed in eukaryotic cells via a specific oligonucleotide-directed enrichment. We demonstrate the applicability of this approach by isolating a cognate RNP bound to the retroviral 5' untranslated region that is composed of DHX9/RNA helicase A.

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Environment

Reliable Method for Assessing Seed Germination, Dormancy, and Mortality under Field Conditions
Brian A. Pace 1, Helen M. Alexander 2, D. Jason Emry 3, Kristin L. Mercer 1
1Department of Horticulture and Crop Science, Ohio State University, 2Department of Ecology and Evolutionary Biology, University of Kansas, 3Department of Biology, Washburn University

Here we present a protocol for assessing seed survivorship, germination and dormancy under field conditions using buried, labeled seed strips and tetrazolium chloride (TZ) viability testing.

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Environment

The Calibration and Use of Capacitance Sensors to Monitor Stem Water Content in Trees
Ashley M. Matheny 1, Steven R. Garrity 2, Gil Bohrer 3
1Department of Geological Sciences, Jackson School of Geosciences, University of Texas at Austin, 2METER Group, Inc., USA, 3Department of Civil, Environmental and Geodetic Engineering, Ohio State University

The hydraulic capacitance of biomass is a key component of the vegetation water budget, which serves as a buffer against short and long-term drought stresses. Here, we present a protocol for the calibration and use of soil moisture capacitance sensors to monitor water content in the stems of large trees.

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Medicine

Ferric Chloride-induced Canine Carotid Artery Thrombosis: A Large Animal Model of Vascular Injury
Allyson L. Huttinger 1, Debra G. Wheeler 1, Surya Gnyawali 2, David Dornbos III 1, Juliana M. Layzer 3, Nicholas Venetos 1, Spencer Talentino 1, Nicholas J. Musgrave 1, Cheyenne Jones 1, Camille Bratton 1, Matthew E. Joseph 4, Chandan Sen 2, Bruce A. Sullenger 3, Shahid M. Nimjee 1
1Department of Neurological Surgery, Ohio State University, 2Department of Surgery, Ohio State University, 3Department of Surgery, Duke University, 4Department of Internal Medicine, Ohio State University

Here, we present the modifications necessary to a well characterized and commonly used small animal ferric chloride-induced (FeCl3) carotid artery injury model for use in a large animal vascular injury model. The resulting model can be utilized for pre-clinical trial assessment of both prophylactic and thrombolytic pharmacological and mechanical interventions.

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Behavior

Novel Object Recognition and Object Location Behavioral Testing in Mice on a Budget
Jiyeon K. Denninger 1, Bryon M. Smith 1, Elizabeth D. Kirby 1,2,3
1Department of Psychology, Ohio State University, 2Department of Neuroscience, Ohio State University, 3Center for Chronic Brain Injury, Ohio State University

Here we provide a protocol which includes comprehensive instructions for the economical establishment of murine object location and novel object recognition behavioral testing, including the design, cost, and construction of required equipment as well as execution of behavioral testing, data collection, and analysis.

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Cancer Research

Detection of Protease Activity by Fluorescent Peptide Zymography
Ameya A. Deshmukh 1,2, Jessica L. Weist 1, Jennifer L. Leight 1,2
1Comprehensive Cancer Center, James Cancer Hospital and Solove Research Center, Ohio State University, 2Department of Biomedical Engineering, College of Engineering, Ohio State University

Here, we present a detailed protocol for a modified zymographic technique in which fluorescent peptides are used as the degradable substrate in place of native proteins. Electrophoresis of biological samples in fluorescent peptide zymograms enables detection of a wider range of proteases than previous zymographic techniques.

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Bioengineering

Measuring Global Cellular Matrix Metalloproteinase and Metabolic Activity in 3D Hydrogels
Abdulaziz S. Fakhouri 1,2,3, Jennifer L. Leight 1,2
1Department of Biomedical Engineering, The Ohio State University, 2The Ohio State University Comprehensive Cancer Center, Arthur G. James Cancer Hospital and Richard J. Solove Research Institute, 3Biomedical Technology Department, King Saud University

Here, a protocol is presented for encapsulating and culturing cells in poly(ethylene glycol) (PEG) hydrogels functionalized with a fluorogenic matrix metalloproteinase (MMP)-degradable peptide. Cellular MMP and metabolic activity are measured directly from the hydrogel cultures using a standard microplate reader.

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JoVE Journal

Quantification of Metal Leaching in Immobilized Metal Affinity Chromatography
Coleman M. Swaim 1, Tyler J. Brittain 1, Daniel R. Marzolf 2, Oleksandr Kokhan 1
1Department of Chemistry and Biochemistry, James Madison University, 2Biophysics Graduate Program, Ohio State University

We present an assay for easy quantification of metals introduced to samples prepared using immobilized metal affinity chromatography. The method uses hydroxynaphthol blue as the colorimetric metal indicator and a UV-Vis spectrophotometer as the detector.

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Engineering

Surgery and Sample Processing for Correlative Imaging of the Murine Pulmonary Valve
Yifei Liu 1, Yong-Ung Lee 2, Tai Yi 2, Ken Wu 3, Cedric Bouchet-Marquis 3, Han Chan 3, Christopher K. Breuer 2, David W. McComb 1
1Center for Electron Microscopy and Analysis, The Department of Materials Science and Engineering, Ohio State University, 2Center for Regenerative Medicine, Nationwide Children’s Hospital, 3Thermo Fisher Scientific

Here, we describe a correlative workflow for the excision, pressurization, fixation, and imaging of the murine pulmonary valve to determine the gross conformation and local extracellular matrix structures.

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Medicine

Targeted Muscle Reinnervation: Surgical Protocol for a Randomized Controlled Trial in Postamputation Pain
Emily Pettersen 1,2,3,4, Paolo Sassu 5, Francesca Alice Pedrini 1,5, Hannes Granberg 1,2, Carina Reinholdt 2,6, Juan Manuel Breyer 7, Aidan Roche 8, Andrew Hart 9,10, Adil Ladak 11, Hollie A. Power 11, Michael Leung 12, Michael Lo 12, Ian Valerio 13, Kyle R. Eberlin 13, Theodore A. Kung 14, Paul Cederna 14, Jason M. Souza 15,16, Oskar Aszmann 17, Jason Ko 18, Gregory A. Dumanian 18, Max Ortiz-Catalan 1,4,19,20
1Center for Bionics and Pain Research, 2Center for Advanced Reconstruction of Extremities, Sahlgrenska University Hospital, 3Department of Electrical Engineering, Chalmers University of Technology, 4Bionics Institute, 5IV Clinica Ortoplastica, IRCCS Istituto Ortopedico Rizzoli, 6Department of Hand Surgery, Institute of Clinical Sciences, Sahlgrenska Academy, University of Gothenburg, Sahlgrenska University Hospital, 7Department of Orthopedic, Hand Unit, Worker Hospital, 8College of Medicine and Veterinary Medicine, The Queen's Medical Research Institute, The University of Edinburgh, 9Canniesburn Plastic Surgery Unit, Glasgow Royal Infirmary, 10College of Medicine, Veterinary & Life Sciences, The University of Glasgow, 11Division of Plastic Surgery, Department of Surgery, Faculty of Medicine and Dentistry, University of Alberta, 12Plastics and Reconstructive Surgery, Dandenong Hospital, Monash Health, 13Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital &, Harvard Medical School, 14Section of Plastic Surgery, Department of Surgery, Michigan Medicine, 15Department of Surgery, Uniformed Services University of the Health Sciences and Walter Reed National Military Medical Center, 16Department of Plastic and Reconstructive Surgery, Ohio State University, 17Clinical Laboratory for Bionic Extremity Reconstruction, Department of Plastic, Reconstructive and Aesthetic Surgery, Medical University Vienna, 18Division of Plastic Surgery, Department of Surgery, Northwestern Feinberg School of Medicine, 19Medical Bionics Department, University of Melbourne, 20Prometei Pain Rehabilitation Center

The protocol outlines the surgical procedure for the treatment of postamputation pain using Targeted Muscle Reinnervation (TMR). TMR will be compared with two other surgical techniques, specifically Regenerative Peripheral Nerve Interface (RPNI) and neuroma excision, followed by immediate burying within muscle under the context of an international, randomized controlled trial.

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