University of Vermont View Institution's Website 21 articles published in JoVE Bioengineering Dissection and Isolation of Region-Specific Decellularized Lung Tissue Evan T. Hoffman1, Isaac D. Downs2, Brad Young2, Loredana Asarian1, Daniel J. Weiss1 1Larner College of Medicine, University of Vermont, 2College of Engineering and Mathematical Sciences, University of Vermont Presented here is a protocol for the isolation of regional decellularized lung tissue. This protocol provides a powerful tool for studying complexities in the extracellular matrix and cell-matrix interactions. Medicine In Vivo Quantification of Hip Arthrokinematics during Dynamic Weight-bearing Activities using Dual Fluoroscopy Penny R. Atkins1,2, Niccolo M. Fiorentino1,3, Andrew E. Anderson1,2,4,5 1Department of Orthopaedics, University of Utah, 2Scientific Computing and Imaging Institute, University of Utah, 3Department of Mechanical Engineering, University of Vermont, 4Department of Biomedical Engineering, University of Utah, 5Department of Physical Therapy, University of Utah Dual fluoroscopy accurately captures in vivo dynamic motion of human joints, which can be visualized relative to reconstructed anatomy (e.g., arthrokinematics). Herein, a detailed protocol to quantify hip arthrokinematics during weight-bearing activities of daily living is presented, including the integration of dual fluoroscopy with traditional skin marker motion capture. Environment Simulating Impacts of Ice Storms on Forest Ecosystems John L. Campbell1, Lindsey E. Rustad1, Charles T. Driscoll2, Ian Halm3, Timothy J. Fahey4, Habibollah Fakhraei5, Peter M. Groffman6,7, Gary J. Hawley8, Wendy Leuenberger9, Paul G. Schaberg10 1Northern Research Station, U.S. Forest Service, Durham, NH, 2Department of Civil and Environmental Engineering, Syracuse University, 3Northern Research Station, U.S. Forest Service, North Woodstock, NH, 4Department of Natural Resources, Cornell University, 5Department of Civil and Environmental Engineering, Southern Illinois University, 6Advanced Science Research Center at the Graduate Center, City University of New York, 7Cary Institute of Ecosystem Studies, 8Rubenstein School of Environment and Natural Resources, University of Vermont, 9Department of Forestry and Natural Resources, University of Kentucky, 10Northern Research Station, U.S. Forest Service, Burlington, VT Ice storms are important weather events that are challenging to study because of difficulties in predicting their occurrence. Here, we describe a novel method for simulating ice storms that involves spraying water over a forest canopy during sub-freezing conditions. Biology Single Cell Durotaxis Assay for Assessing Mechanical Control of Cellular Movement and Related Signaling Events Kathryn V. Svec1,2,3, Johnathan B Patterson1,2,3, Nyla Naim1,2,3, Alan K Howe1,2,3 1Department of Pharmacology, University of Vermont Larner College of Medicine, 2University of Vermont Cancer Center, 3Department of Molecular Physiology and Biophysics, University of Vermont Larner College of Medicine Mechanical forces are important for controlling cell migration. This protocol demonstrates the use of elastic hydrogels that can be deformed using a glass micropipette and a micromanipulator to stimulate cells with a local stiffness gradient to elicit changes in cell structure and migration. Environment Measuring Phosphorus Release in Laboratory Microcosms for Water Quality Assessment Eric O. Young1, Donald S. Ross2, Jessica Sherman1 1United States Department of Agriculture, Agricultural Research Service (USDA-ARS), Institute for Environmentally Integrated Dairy Management, 2Department of Plant and Soil Science, University of Vermont Accurate quantification of phosphorus (P) desorption potential in saturated soils and sediments is important for P modeling and transport mitigation efforts. To better account for in situ soil-water redox dynamics and P mobilization under prolonged saturation, a simple approach was developed based on repeated sampling of laboratory microcosms. Biology Applications of Spatio-temporal Mapping and Particle Analysis Techniques to Quantify Intracellular Ca2+ Signaling In Situ Bernard T. Drumm1, Grant W. Hennig2, Salah A. Baker1, Kenton M. Sanders1 1Department of Physiology and Cell Biology, University of Nevada Reno School of Medicine, 2Department of Pharmacology, The Robert Larner, M.D. College of Medicine, University of Vermont Genetically encoded Ca2+ indicators (GECIs) have radically changed how in situ Ca2+ imaging is performed. To maximize data recovery from such recordings, appropriate analysis of Ca2+ signals is required. The protocols in this paper facilitate the quantification of Ca2+ signals recorded in situ using spatiotemporal mapping and particle-based analysis. Biology A Plate Competition Assay As a Quick Preliminary Assessment of Disease Suppression Deborah A. Neher1, Thomas R. Weicht1 1Department of Plant and Soil Science, University of Vermont Presented is a protocol for a plate competition assay to identify whether a specific compost is likely to contain bacteria and fungi that suppress growth of Rhizoctonia solani. Neuroscience Ex Vivo Imaging of Cell-specific Calcium Signaling at the Tripartite Synapse of the Mouse Diaphragm Dante J. Heredia1, Grant W. Hennig2, Thomas W. Gould1 1Department of Physiology and Cell Biology, School of Medicine, University of Nevada, 2Department of Pharmacology, Larner College of Medicine, University of Vermont Here we present a protocol to image calcium signaling in populations of individual cell types at the murine neuromuscular junction. Chemistry Deposition of Porous Sorbents on Fabric Supports Brandy J. Johnson1, Brian J. Melde1, Martin H. Moore1, Jenna R. Taft2 1Center for Bio/Molecular Science & Engineering, Naval Research Laboratory, 2Department of Chemistry, University of Vermont This report details a microwave-initiated approach for deposition of porphyrin functionalized porous organosilicate sorbents on a cotton fabric and demonstrates reduction in 2-chloroethyl ethyl sulfide (CEES) transport through the fabric resulting from this treatment. Medicine Cavernous Nerve Stimulation and Recording of Intracavernous Pressure in a Rat Morten Hox1, Travis Mann-Gow2, Lars Lund3,4, Peter Zvara1,4 1Biomedical Laboratory and the Research Unit of Urology, Department of Clinical Research, University of Southern Denmark, 2Department of Surgery, University of Vermont, 3Research Unit of Urology, Department of Clinical Research, University of Southern Denmark, 4Department of Urology, Odense University Hospital This study describes a simplified surgical procedure and technique for performing cavernous nerve stimulation with the isolation of the nerve-electrode complex using silicone glue and intracavernous pressure measurement. Behavior A Semantic Priming Event-related Potential (ERP) Task to Study Lexico-semantic and Visuo-semantic Processing in Autism Spectrum Disorder Emily L. Coderre1 1Department of Communication Sciences and Disorders, University of Vermont This paper describes a semantic priming ERP task using within-modality pairs of pictures and words to investigate semantic processing in individuals with autism spectrum disorder (ASD). Medicine Evaluating the Procedure for Performing Awake Cystometry in a Mouse Model Travis K. Mann-Gow*1, Troy R. Larson*1, Chrissie T. Wøien2, Thomas M. Andersen2, Karl-Erik Andersson3,4, Peter Zvara1,2 1Department of Surgery, University of Vermont, 2Department of Urology and Biomedical Laboratory, University of Southern Denmark, 3Wake Forest Institute for Regenerative Medicine, Wake Forest University, 4Institute for Clinical Medicine, Aarhus University This study describes the surgical procedures and experimental techniques for performing awake cystometry in a freely moving mouse. In addition, it provides experimental evidence to support its optimization and standardization. Environment Methods of Soil Resampling to Monitor Changes in the Chemical Concentrations of Forest Soils Gregory B. Lawrence1, Ivan J. Fernandez2, Paul W. Hazlett3, Scott W. Bailey4, Donald S. Ross5, Thomas R. Villars6, Angelica Quintana7, Rock Ouimet8, Michael R. McHale1, Chris E. Johnson9, Russell D. Briggs10, Robert A. Colter11, Jason Siemion1, Olivia L. Bartlett12, Olga Vargas13, Michael R. Antidormi1, Mary M. Koppers9 1New York Water Science Center, U.S. Geological Survey, 2School of Forest Resources, University of Maine, 3Natural Resources Canada, Canadian Forest Service, 4Northern Research Station, U.S. Forest Service, 5Department of Plant and Soil Science, University of Vermont, 6Ottauquechee NRCD, USDA Natural Resources Conservation Service, 7Green Mountain National Forest, U.S. Forest Service, 8Direction de la Recherche Forestière, Ministère du Québec, 9Department of Civil and Environmental Engineering, Syracuse University, 10Division of Environmental Science, SUNY College of Environmental Science and Forestry, 11White Mountain National Forest, U.S. Forest Service, 12Natural Resources and Earth System Sciences, University of New Hampshire, 13Greenwich, NY Field Office, USDA Natural Resources Conservation Service Repeated soil sampling has recently been shown to be an effective way to monitor forest soil change over years and decades. To support its use, a protocol is presented that synthesizes the latest information on soil resampling methods to aid in the design and implementation of successful soil monitoring programs. Neuroscience Isolation and Cannulation of Cerebral Parenchymal Arterioles Paulo W. Pires1, Fabrice Dabertrand2, Scott Earley1 1Department of Pharmacology, University of Nevada School of Medicine, 2Department of Pharmacology, University of Vermont College of Medicine This manuscript describes a simple and reproducible protocol for isolation of intracerebral arterioles (a group of blood vessels encompassing parenchymal arterioles, penetrating arterioles and pre-capillary arterioles) from mice, to be used in pressure myography, immunofluorescence, biochemistry, and molecular studies. Immunology and Infection Highly Sensitive Assay for Measurement of Arenavirus-cell Attachment Joseph P. Klaus1, Jason Botten1,2 1Department of Medicine, Division of Immunobiology, University of Vermont, 2Department of Microbiology & Molecular Genetics, University of Vermont The first step in the arenavirus life cycle is attachment of viral particles to host cells. We report a quantitative (q)RT-PCR-based assay for ultrasensitive detection and quantitation of arenavirus attachment events. Behavior Place and Response Learning in the Open-field Tower Maze Olga Lipatova1, Matthew M. Campolattaro1, Donna J. Toufexis2, Erin A. Mabry1 1Department of Psychology, Christopher Newport University, 2Department of Psychological Science, University of Vermont The Open-field Tower Maze (OFTM) is a maze created to study the behavioral and neural mechanisms of spatial learning (e.g., place- or response-learning) in rats. This maze is especially useful for experimenters who want to use a non-stressful maze paradigm to investigate spatial learning in their research. Biology Measurement of Smooth Muscle Function in the Isolated Tissue Bath-applications to Pharmacology Research Brian Jespersen1, Nathan R. Tykocki2, Stephanie W. Watts1, Peter J. Cobbett1 1Department of Pharmacology and Toxicology, Michigan State University, 2Department of Pharmacology, University of Vermont College of Medicine This protocol describes the measurement of isometric contraction in an isolated smooth muscle preparation, using an isolated tissue bath system and computer-based data acquisition. Bioengineering Characterizing Bacterial Volatiles using Secondary Electrospray Ionization Mass Spectrometry (SESI-MS) Heather D. Bean1, Jiangjiang Zhu1, Jane E. Hill1 1School of Engineering, University of Vermont Secondary electrospray ionization mass spectrometry (SESI-MS) enables the detection of volatile organic compounds (VOCs) without the need for any sample pretreatment. This protocol provides instructions for the rapid (within minutes) characterization of bacterial VOCs using SESI-MS. Biology High-Throughput Measurement and Classification of Organic P in Environmental Samples Nicholas R. Johnson1, Jane E. Hill1 1School of Engineering, University of Vermont This protocol describes a high-throughput method of enzymatic hydrolysis that utilizes a microplate reader to measure and classify soil phosphorus as P monoesters, P diesters and inorganic P. Up to 96 samples can be measured at one time in a standard laboratory. Biology Microarray Analysis for Saccharomyces cerevisiae Scott Tighe1, Tim Hunter1, Pat Reed1, Janet Murray1 1Vermont Genetics Network, The University of Vermont In this protocol, gene expression in yeast (Saccharomyces cerevisiae) is changed after exposure to oxidative stress induced by the addition of hydrogen peroxide (H2O2), an oxidizing agent. Biology Gastrointestinal Motility Monitor (GIMM) Jill M. Hoffman1, Elice M. Brooks1, Gary M. Mawe1 1Department of Anatomy and Neurobiology, The University of Vermont Evaluation of colonic motility in the guinea pig distal colon with the Gastrointestinal Motility Monitor (GIMM) is a straightforward and simple to learn approach to quantitatively evaluate propulsive motility in the gastrointestinal tract.