University of Texas, Arlington View Institution's Website 9 articles published in JoVE Biology Generating Transposon Insertion Libraries in Gram-Negative Bacteria for High-Throughput Sequencing Misha I. Kazi1, Richard D. Schargel1, Joseph M. Boll1 1Department of Biology, University of Texas at Arlington We describe a method to generate saturating transposon mutant libraries in Gram-negative bacteria and subsequent preparation of DNA amplicon libraries for high-throughput sequencing. As an example, we focus on the ESKAPE pathogen, Acinetobacter baumannii, but this protocol is amenable to a wide range of Gram-negative organisms. Bioengineering Light-sheet Fluorescence Microscopy for the Study of the Murine Heart Yichen Ding1, Zachary Bailey2, Victoria Messerschmidt2, Jun Nie3, Richard Bryant2, Sandra Rugonyi4, Peng Fei3, Juhyun Lee1,2, Tzung K. Hsiai1 1Department of Bioengineering, University of California Los Angeles, 2Department of Bioengineering, University of Texas at Arlington, 3School of Optical and Electronic Information, Huazhong University of Science and Technology, 4Department of Biomedical Engineering, OSHU This study uses a dual-sided illumination light-sheet fluorescence microscopy (LSFM) technique combined with optical clearing to study the murine heart. Bioengineering Light-sheet Fluorescence Microscopy to Capture 4-Dimensional Images of the Effects of Modulating Shear Stress on the Developing Zebrafish Heart Victoria Messerschmidt*1, Zachary Bailey*1, Kyung In Baek2, Yichen Ding2, Jeffrey J. Hsu2, Richard Bryant1, Rongsong Li3, Tzung K. Hsiai2, Juhyun Lee1 1Department of Bioengineering, The University of Texas at Arlington, 2Department of Medicine (Cardiology) and Bioengineering, UCLA, 3College of Health Science and Environmental Engineering, Shenzhen Technology University Here, we present a protocol to visualize developing hearts in zebrafish in 4-Dimensions (4-D). 4-D imaging, via light-sheet fluorescence microscopy (LSFM), takes 3-Dimensional (3-D) images over time, to reconstruct developing hearts. We show qualitatively and quantitatively that shear stress activates endocardial Notch signaling during chamber development, which promotes cardiac trabeculation. Medicine Skeletal Muscle Neurovascular Coupling, Oxidative Capacity, and Microvascular Function with 'One Stop Shop' Near-infrared Spectroscopy Ryan Rosenberry1, Susie Chung1, Michael D. Nelson1 1Applied Physiology and Advanced Imaging Laboratory, Department of Kinesiology, University of Texas at Arlington Here, we describe a simple, non-invasive approach using near-infrared spectroscopy to assess reactive hyperemia, neurovascular coupling and skeletal muscle oxidative capacity in a single clinic or laboratory visit. Bioengineering Patterning Bioactive Proteins or Peptides on Hydrogel Using Photochemistry for Biological Applications Taylor B. Dorsey1,2,3, Alexander Grath1,2, Cancan Xu4, Yi Hong4, Guohao Dai1,2,3 1Department of Biomedical Engineering, Rensselaer Polytechnic Institute, 2Center for Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute, 3Department of Bioengineering, Northeastern University, 4Department of Bioengineering, University of Texas at Arlington In this method, we use photopolymerization and click chemistry techniques to create protein or peptide patterns on the surface of polyethylene glycol (PEG) hydrogels, providing immobilized bioactive signals for the study of cellular responses in vitro. Immunology and Infection Detection of Human Leukocyte Antigen Biomarkers in Breast Cancer Utilizing Label-free Biosensor Technology Jon A. Weidanz1, Krysten L. Doll1, Soumya Mohana-Sundaram1, Timea Wichner1, Devin B. Lowe1, Susanne Gimlin2, Debra Wawro Weidanz2, Robert Magnusson2,3, Oriana E. Hawkins1 1Experimmune, A Center for Immunotherapeutic Development, Department of Immunotherapeutics and Biotechnology, Texas Tech University Health Sciences Center, 2Resonant Sensors Incorporated, 3University of Texas Arlington Intact class I HLA/peptide complexes are shed by cancer cells, representing a potential relevant cancer biomarker. Utilizing label-free sensor technology and T-cell receptor mimicking monoclonal antibodies, detection of shed MIF/HLA-A*02:01 complexes in MDA-MB-231 cell supernatants, spiked human serum, and patient plasma is demonstrated, enabling development of a novel cancer diagnostic platform. Engineering Micropunching Lithography for Generating Micro- and Submicron-patterns on Polymer Substrates Anirban Chakraborty1, Xinchuan Liu1, Cheng Luo1 1Mechanical and Aerospace Engineering, University of Texas at Arlington A micropunching lithography approach is developed to generate micro- and submicron-patterns on top, sidewall and bottom surfaces of polymer substrates. It overcomes the obstacles of patterning conducting polymers and generating sidewall patterns. This method allows rapid fabrication of multiple features and is free of aggressive chemistry. Biology An Introduction to Worm Lab: from Culturing Worms to Mutagenesis Jyotiska Chaudhuri*1, Manish Parihar*1, Andre Pires-daSilva1 1Department of Biology, University of Texas at Arlington Screening for mutants with phenotypic defects is a straightforward method for identifying genes that function in a given biological process. In this article we describe how to culture free living worms (e.g., Pristionchus pacificus) in the laboratory and show two different mutagenesis methods, EMS and TMP/UV. Immunology and Infection Assessing Stomatal Response to Live Bacterial Cells using Whole Leaf Imaging Reejana Chitrakar1, Maeli Melotto1 1Biology, University of Texas at Arlington We have developed a simple and reproducible protocol to access stomatal response to live bacteria. This method minimizes wounding and manipulation of the leaf as compared to the use of epidermal peels reported previously.
