Utah State University 16 articles published in JoVE Biology Isolation of Primary Porcine Retinal Pigment Epithelial Cells for In Vitro Modeling Chase A. Paterson1, Dillon Weatherston1, Teren Teeples1, Elizabeth Vargis1 1Department of Biological Engineering, Utah State University This protocol outlines the procedure for obtaining and culturing primary retinal pigment epithelial (RPE) cells from locally sourced porcine eyes. These cells serve as a high-quality alternative to stem cells and are suitable for in vitro retinal research. Environment Big Box Biochar Kiln Operation and Best Practices Darren J. McAvoy1 1Utah State University The Utah Biomass Resources Group (UBRG) has scaled-up simple biochar kilns in an innovative approach to hazardous fuel reduction and biochar production using metal boxes, called Big Box kilns, that allow for in-woods biochar production. This article outlines Big Box biochar kiln operation and best practices. Developmental Biology Use of Bisection to Reduce Mitochondrial DNA in the Bovine Oocyte Laura Adams*1, Ying Liu*1, Barbara Durrant2, Elena Ruggeri2, Carly Young2, Rebecca Krisher3, Irina Polejaeva1 1Department of Animal, Dairy, and Veterinary Sciences, Utah State University, 2Reproductive Sciences, Beckman Center for Conservation Research, San Diego Zoo Wildlife Alliance, 3Genus PLC Here, we present a protocol to significantly reduce the mitochondrial DNA copy numbers in a bovine oocyte (P < 0.0001). This method utilizes centrifugation and bisection to substantially reduce oocyte mitochondria and may allow for an increased chance of development in the reconstructed interspecies somatic cell nuclear transfer embryos. Biology A 3D Printed Pollen Trap for Bumble Bee (Bombus) Hive Entrances Houston J. Judd1,2, Craig Huntzinger2, Ricardo Ramirez1, James P. Strange2,3 1Department of Biology, Utah State University, 2USDA-ARS Pollinating Insect Research Unit, 3Department of Entomology, The Ohio State University We present a non-lethal and automated mechanism to collect pollen from bumble bee (Bombus) workers returning to a hive. Instructions for producing, preparing, installing and using the devices are included. By using 3D-printed objects, modification to the design was timely, efficient and allowed for quick turnaround for testing. Behavior A Modified Lean and Release Technique to Emphasize Response Inhibition and Action Selection in Reactive Balance David A.E. Bolton1, Mahmoud Mansour2 1Department of Kinesiology & Health Science, Utah State University, 2Department of Electrical & Computer Engineering, Utah State University Here we offer a protocol that allows the user to selectively change affordances and/or constraints on movements that are relevant for recovering balance after postural perturbation. Behavior A Cross-Disciplinary and Multi-Modal Experimental Design for Studying Near-Real-Time Authentic Examination Experiences Idalis Villanueva1, Jenefer Husman3, Darcie Christensen1, Kate Youmans1, Md Tarique Khan1, Paul Vicioso2, Shawn Lampkins3, Matthew C. Graham3 1Department of Engineering Education, College of Engineering, Utah State University, 2Department of Electrical and Computer Engineering, College of Engineering, Utah State University, 3Department of Educational Psychology, College of Education, University of Oregon An experimental design was developed to investigate the real-time influences of an examination experience to assess the emotional realities students experience in higher education settings and tasks. This design is the result of a cross-disciplinary (e.g., educational psychology, biology, physiology, engineering) and multi-modal (e.g., salivary markers, surveys, electrodermal sensor) approach. Bioengineering Material Formation of Recombinant Spider Silks through Aqueous Solvation using Heat and Pressure Justin A. Jones1, Thomas I. Harris1, Brianne E. Bell1, Paula F. Oliveira1 1Department of Biology, Utah State University Here, we present a protocol to produce water soluble recombinant spider silk protein solutions and the material forms that can be formed from those solutions. Engineering Large Area Substrate-Based Nanofabrication of Controllable and Customizable Gold Nanoparticles Via Capped Dewetting Amir Behbahanian1, Nicholas A. Roberts1 1Mechanical and Aerospace Engineering, Utah State University This protocol details a novel nano-manufacturing technique that can be used to make controllable and customizable nanoparticle films over large areas based on the self-assembly of dewetting of capped metal films. Developmental Biology Orthotopic Ovarian Transplantation Procedures to Investigate the Life- and Health-span Influence of Ovarian Senescence in Female Mice Jeffrey B. Mason1, Kate C. Parkinson1, Tracy L. Habermehl1 1Department of Animal, Dairy and Veterinary Sciences, Center for Integrated BioSystems, School of Veterinary Medicine, Utah State University Described here are orthotopic ovarian transplantation protocols for modifying ovarian influence on the physiology of the recipient mouse. This surgical procedure is used to expose female mice to various types of ovarian influence and may include heterochronic transplantations and transplantation of ovaries that have undergone various ex vivo treatments. Bioengineering Production of Genetically Engineered Golden Syrian Hamsters by Pronuclear Injection of the CRISPR/Cas9 Complex Rong Li*1, Jinxin Miao*1,2, Zhiqiang Fan1, SeokHwan Song3, Il-keun Kong3,4, Yaohe Wang2,5, Zhongde Wang1 1Department of Animal, Dairy, and Veterinary Sciences, Utah State University, 2National Centre for International Research in Cell and Gene Therapy, Sino-British Research Centre, School of Basic Medical Sciences, Zhengzhou University, 3Department of Animal Science Division of Applied Life Science (BK21 Plus), Gyeongsang National University, 4Institute of Agriculture and Life Science, Gyeongsang National University, 5Centre for Molecular Oncology, Barts Cancer Institute, Queen Mary University of London Pronuclear (PN) injection of the clustered regularly interspaced short palindromic repeats (CRISPR) and CRISPR-associated protein-9 nuclease (CRISPR/Cas9) system is a highly efficient method for producing genetically engineered golden Syrian hamsters. Herein, we describe the detailed PN injection protocol for the production of gene knockout hamsters with the CRISPR/Cas9 system. Biochemistry Fabricating a UV-Vis and Raman Spectroscopy Immunoassay Platform Cynthia Hanson*1, Nathan D. Israelsen*1, Michael Sieverts1, Elizabeth Vargis1 1Biological Engineering Department, Utah State University Nanoparticle-based optical probes have been designed as a vehicle for detecting antigens using Raman and UV-Vis spectroscopy. Here we describe a protocol for preparing such probes for a UV-Vis/Raman spectroscopy immunoassay in such a way to incorporate future multiplexing capabilities. Behavior Utilizing Electroencephalography Measurements for Comparison of Task-Specific Neural Efficiencies: Spatial Intelligence Tasks Benjamin J. Call1, Wade Goodridge1, Idalis Villanueva1, Nicholas Wan2, Kerry Jordan2 1Department of Engineering Education, Utah State University, 2Department of Psychology, Utah State University This manuscript describes an approach to measure neural activity of humans while solving spatially focused engineering problems. The electroencephalogram methodology helps interpret beta brain wave measurements in terms of neural efficiency, with the aim of ultimately enabling comparisons of task performance both between problem types and between participants. Behavior Use of Galvanic Skin Responses, Salivary Biomarkers, and Self-reports to Assess Undergraduate Student Performance During a Laboratory Exam Activity Idalis Villanueva1, Maria Valladares1, Wade Goodridge1 1Department of Engineering Education, Utah State University Research on undergraduate students' academic achievement emotions have primarily relied on self-reports in laboratory settings. Studies rarely include bio-physiological measures to these self-reports. This protocol will describe a methodology that integrates self-reports with and bio-physiological measures to assess student response and performance during a laboratory examination activity. Behavior Measuring Delay Discounting in Humans Using an Adjusting Amount Task Charles C.J. Frye1, Ann Galizio1, Jonathan E. Friedel1, W. Brady DeHart1, Amy L. Odum1 1Department of Psychology, Utah State University Delay discounting refers to a decline in the value of a reward when it is delayed relative to when it is immediately available. We outline a computer-based delay discounting task that is easy to implement and allows for the quantification of the degree of delay discounting in human participants. Immunology and Infection Bacterial Artificial Chromosomes: A Functional Genomics Tool for the Study of Positive-strand RNA Viruses Sang-Im Yun1, Byung-Hak Song1, Jin-Kyoung Kim1, Young-Min Lee1 1Department of Animal, Dairy, and Veterinary Sciences, Utah Science Technology and Research, College of Agriculture and Applied Sciences, Utah State University Here, a protocol is presented for creating an infectious bacterial artificial chromosome containing a full-length cDNA of the positive-strand genomic RNA of Japanese encephalitis virus. This protocol can be used to construct a functional cDNA of other positive-strand RNA viruses, making it a powerful genomic tool for studying virus biology. Environment Quantification of Heavy Metals and Other Inorganic Contaminants on the Productivity of Microalgae Katerine Napan1, Derek Hess1, Brian McNeil1, Jason C. Quinn1 1Mechanical and Aerospace Engineering, Utah State University Integration of microalgal cultivation with industrial flue gas will ultimately introduce heavy metals and other inorganic compounds into the growth media. This study presents a procedure used to determine the end fate and impact of heavy metals and inorganic contaminants on the growth of Nannochloropsis salina grown in photobioreactors.
