Oklahoma State University

10 ARTICLES PUBLISHED IN JoVE

Immunology and Infection

Katherine M. Kocan¹, Edmour Blouin¹, José de la Fuente^1,2

¹Department of Veterinary Pathobiology, Center for Veterinary Health Sciences, Oklahoma State University, ²(CSIC-UCLM-JCCM), Instituto de Investigación en Recursos Cinegéticos IREC

A method for RNA interference (RNAi) by injection of dsRNA into unfed ticks is described. RNAi is the most widely used gene-silencing technique in ticks where the use of other methods of genetic manipulation has been limited.

Neuroscience

Electrophysiological Recording in the Brain of Intact Adult Zebrafish

Lindsey Johnston¹, Rebecca E. Ball², Seth Acuff³, John Gaudet⁴, Andrew Sornborger⁵, James D. Lauderdale²

¹Department of Genetics, University of Georgia, ²Department of Cellular Biology, University of Georgia, ³College of Engineering, Architecture and Technology, Oklahoma State University, ⁴University of Georgia, ⁵Department of Mathematics, University of California, Davis

This paper describes how an adult zebrafish can be immobilized, intubated, and used for in vivo electrophysiological experiments to allow recordings and manipulation of neural activity in an intact animal.

Biology

Single-cell Microfluidic Analysis of Bacillus subtilis

Matthew T. Cabeen¹, Richard Losick²

¹Department of Microbiology and Molecular Genetics, Oklahoma State University, ²Department of Molecular and Cellular Biology, Harvard University

We present a method for the microfluidic analysis of individual bacterial cell lineages using Bacillus subtilis as an example. The method overcomes shortcomings of traditional analytical methods in microbiology by allowing observation of hundreds of cell generations under tightly controllable and uniform growth conditions.

Immunology and Infection

Combining Analysis of DNA in a Crude Virion Extraction with the Analysis of RNA from Infected Leaves to Discover New Virus Genomes

Jeanmarie Verchot¹, Aastha Thapa², Dulanjani Wijayasekara³, Peter R. Hoyt⁴

¹Texas A&M Agrilife Center at Dallas, ²Noble Research Center, Oklahoma State University, ³Department of Biology, College of Engineering and Natural Sciences, The University of Tulsa, ⁴Bioinformatics and Genomics Core Facility, Department of Biochemistry and Molecular Biology, Oklahoma State University

Here we present a new approach to identify plant viruses with double-strand DNA genomes. We use standard methods to extract DNA and RNA from infected leaves and carry out next-generation sequencing. Bioinformatic tools assemble sequences into contigs, identify contigs representing virus genomes and assign genomes to taxonomic groups.

Immunology and Infection

Three-dimensional Imaging of Bacterial Cells for Accurate Cellular Representations and Precise Protein Localization

Benjamin P. Bratton², Brody Barton¹, Randy M. Morgenstein¹

¹Department of Microbiology and Molecular Genetics, Oklahoma State University, ²Department of Molecular Biology and Lewis-Sigler Institute of Integrative Genomics, Princeton University

This protocol explains how to prepare and mount bacterial samples for live three-dimensional imaging and how to reconstruct the three-dimensional shape of E. coli from those images.

Engineering

Terahertz Imaging and Characterization Protocol for Freshly Excised Breast Cancer Tumors

Nagma Vohra^*1, Tyler Bowman^*1, Keith Bailey², Magda El-Shenawee¹

¹Department of Electrical Engineering, University of Arkansas, ²Oklahoma Animal Disease Diagnostic Laboratory, Oklahoma State University

Freshly excised human breast cancer tumors are characterized with terahertz spectroscopy and imaging following fresh tissue handling protocols. Tissue positioning is taken into consideration to enable effective characterization while providing analysis in a timely manner for future intraoperative applications.

JoVE Journal

Interactive Molecular Model Assembly with 3D Printing

Elham Fazelpour¹, Christopher J. Fennell¹

¹Department of Chemistry, Oklahoma State University

Physical modeling of microscopic systems helps obtain insights that are difficult to gain by other means. To facilitate the construction of physical molecular models, we demonstrate how 3D printing can be used to assemble functional macroscopic models that capture qualities of molecular systems in a tactile way.

Neuroscience

Coherent Anti-Stokes Raman Spectroscopy (CARS) Application for Imaging Myelination in Brain Slices

Elizabeth A. McCullagh¹, Shani Poleg², Dominik Stich³, Radu Moldovan^*3, Achim Klug^*2

¹Department of Integrative Biology, Oklahoma State University, ²Department of Physiology and Biophysics, University of Colorado Anschutz, ³Advanced Light Microscopy Core, University of Colorado Anschutz

Visualizing myelination is an important goal for many researchers studying the nervous system. CARS is a technique that is compatible with immunofluorescence that can natively image lipids within tissue such as the brain illuminating specialized structures such as myelin.

Immunology and Infection

Zebrafish Animal Model for the Study of Allergic Reactions in Response to Tick Saliva Biomolecules

Marinela Contreras¹, Almudena González-García¹, José de la Fuente^1,2

¹SaBio. Instituto de Investigación en Recursos Cinegéticos IREC (CSIC-UCLM-JCCM), ²Department of Veterinary Pathobiology, College of Veterinary Medicine, Oklahoma State University

Here, zebrafish (Danio rerio) is used as a model to study allergic reactions and immune responses related to alpha-Gal syndrome (AGS) by evaluating allergic reactions to tick saliva and mammalian meat consumption.

Biology

High-Resolution Fluoro-Respirometry of Equine Skeletal Muscle

Michael Scott Davis¹, Montana Renae Barrett¹

¹Department of Physiological Sciences, Oklahoma State University

Horses have an exceptional aerobic exercise capacity, making equine skeletal muscle an important tissue for both the study of equine exercise physiology as well as mammalian mitochondrial physiology. This article describes techniques for the comprehensive assessment of mitochondrial function in equine skeletal muscle.