Naval Research Laboratory View Institution's Website 16 articles published in JoVE Biology High-throughput Confocal Imaging of Quantum Dot-Conjugated SARS-CoV-2 Spike Trimers to Track Binding and Endocytosis in HEK293T Cells Bruce Nguyen Tran1, Eunkeu Oh2, Kimihiro Susumu2,3, Mason Wolak2, Kirill Gorshkov1 1National Center for Advancing Translational Sciences, 2Optical Sciences Division, Code 5600, Naval Research Laboratory, 3Jacobs Corporation In this protocol, quantum dots conjugated to recombinant SARS-CoV-2 spike enable cell-based assays to monitor spike binding to hACE2 at the plasma membrane and subsequent endocytosis of the bound proteins into the cytoplasm. Immunology and Infection Analysis of Group IV Viral SSHHPS Using In Vitro and In Silico Methods Xin Hu1, Jaimee R. Compton2, Patricia M. Legler2 1National Center for Advancing Translational Sciences, National Institutes of Health, 2United States Naval Research Laboratory We present a general protocol for identifying short stretches of homologous host-pathogen protein sequences (SSHHPS) embedded in the viral polyprotein. SSHHPS are recognized by viral proteases and direct the targeted destruction of specific host proteins by several Group IV viruses. 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. Chemistry Characterizing Electron Transport through Living Biofilms Matthew Yates1, Sarah Strycharz-Glaven1, Joel Golden1, Jared Roy1,2, Stanislav Tsoi3, Jeffrey Erickson1, Mohamed El-Naggar4, Scott Calabrese Barton5, Leonard Tender1 1Center for Bio/Molecular Science and Engineering, Naval Research Laboratory, 2George Mason University, 3Chemistry Division, Naval Research Laboratory, 4Departments of Physics, Biological Sciences, and Chemistry, University of Southern California, 5Department of Chemical Engineering and Materials Science, Michigan State University A protocol for measuring electrical conductivity of living microbial biofilms under physiologically relevant conditions is presented. Engineering Atomic Layer Deposition of Vanadium Dioxide and a Temperature-dependent Optical Model Marc Currie1, Michael A. Mastro1, Virginia D. Wheeler1 1Naval Research Laboratory Thin films (100-1000 Å) of vanadium dioxide (VO2) were created by atomic-layer deposition (ALD) on sapphire substrates. Following this, the optical properties were characterized through the metal-insulator transition of VO2. From the measured optical properties, a model was created to describe the tunable refractive index of VO2. Engineering A Novel Method for In Situ Electromechanical Characterization of Nanoscale Specimens Russell C. Reid1,2, Alberto Piqué1, Wonmo Kang1,3 1Materials Science and Technology Division, US Naval Research Laboratory, 2American Society for Engineering Education—Naval Research Laboratory (ASEE-NRL), 3Leidos Corporation Isolating electrical and thermal effects on electrically assisted deformation (EAD) is very difficult using macroscopic samples. Metallic sample micro- and nanostructures together with a custom test procedure have been developed to evaluate the impact of applied current on the formation without joule heating and evolution of dislocations on these samples. Bioengineering Targeted Plasma Membrane Delivery of a Hydrophobic Cargo Encapsulated in a Liquid Crystal Nanoparticle Carrier Okhil K. Nag1, Jawad Naciri1, Eunkeu Oh2,3, Christopher M. Spillmann1, James B. Delehanty1 1Center for Biomolecular Science and Engineering, Naval Research Laboratory, 2Optical Sciences Division, Naval Research Laboratory, 3Sotera Defense Solutions A liquid crystal nanoparticle (LCNP) nanocarrier is exploited as a vehicle for the controlled delivery of a hydrophobic cargo to the plasma membrane of living cells. Engineering Plasma-assisted Molecular Beam Epitaxy of N-polar InAlN-barrier High-electron-mobility Transistors Matthew T. Hardy1, David F. Storm2, D. Scott Katzer2, Brian P. Downey2, Neeraj Nepal2, David J. Meyer2 1NRC Postdoctoral Scholar, Naval Research Laboratory, 2Electronics Science and Technology Division, Naval Research Laboratory Molecular beam epitaxy is used to grow N-polar InAlN-barrier high-electron-mobility transistors (HEMTs). Control of the wafer preparation, layer growth conditions and epitaxial structure results in smooth, compositionally homogeneous InAlN layers and HEMTs with mobility as high as 1,750 cm2/V∙sec. Biochemistry Directed Protein Packaging within Outer Membrane Vesicles from Escherichia coli: Design, Production and Purification Nathan J. Alves1,2, Kendrick B. Turner1, Scott A. Walper1 1Center for Bio/Molecular Science & Engineering, Naval Research Laboratory, 2Department of Emergency Medicine, Indiana University of School of Medicine A protocol for the production, purification, and use of enzyme packaged outer membrane vesicles (OMV) providing for enhanced enzyme stability for implementation across diverse applications is presented. Engineering Fabrication of Fully Solution Processed Inorganic Nanocrystal Photovoltaic Devices Troy K. Townsend1, Dario Durastanti1, William B. Heuer2, Edward E. Foos3, Woojun Yoon4, Joseph G. Tischler4 1 This protocol describes the synthesis and solution deposition of inorganic nanocrystals layer by layer to produce thin film electronics on non-conductive surfaces. Solvent-stabilized inks can produce complete photovoltaic devices on glass substrates via spin and spray coating following post-deposition ligand exchange and sintering. Engineering Laser-induced Forward Transfer of Ag Nanopaste Eric Breckenfeld1, Heungsoo Kim2, Raymond C. Y. Auyeung2, Alberto Piqué2 1National Research Council Research Associates Program, Naval Research Laboratory, 2Materials Science and Technology Division, Naval Research Laboratory We demonstrate the use of the Laser-induced forward transfer technique (LIFT) for the printing of high-viscosity Ag paste. This technique offers a simple, low temperature, robust process for non-lithographically printing microscale 2D and 3D structures. Bioengineering A Label-free Technique for the Spatio-temporal Imaging of Single Cell Secretions Deepa Raghu1, Joseph A. Christodoulides1, James B. Delehanty2, Jeff M. Byers1, Marc P. Raphael1 1Materials Science and Technology, Naval Research Laboratory, 2Center for Bio/Molecular Science and Engineering, Naval Research Laboratory Inter-cellular communication is critical for controlling various physiological activities within and outside the cell. This paper describes a protocol for measuring the spatio-temporal nature of single cell secretions. To achieve this, a multidisciplinary approach is used which integrates label-free nanoplasmonic sensing with live cell imaging. Engineering Formation of Thick Dense Yttrium Iron Garnet Films Using Aerosol Deposition Scooter D. Johnson1, Evan R. Glaser2, Fritz J. Kub1, Charles R. Eddy, Jr.1 1Power Electronics Branch, Naval Research Laboratory, 2Physics of Electronic Materials Branch, Naval Research Laboratory This report describes the use of a custom-built system to perform aerosol deposition of thick films of yttrium iron garnet onto sapphire substrates at RT. The deposited films are characterized using scanning electron microscopy, profilometry, and ferromagnetic resonance to give a representative overview of the capabilities of the technique. Chemistry Quantitative Detection of Trace Explosive Vapors by Programmed Temperature Desorption Gas Chromatography-Electron Capture Detector Christopher R. Field1, Adam Lubrano2, Morgan Woytowitz2, Braden C. Giordano3, Susan L. Rose-Pehrsson4 1Chemical Sensing & Fuel Technology, Chemistry Division, U.S. Naval Research Laboratory, 2NOVA Research, Inc., 3Bio/Analytical Chemistry, Chemistry Division, U.S. Naval Research Laboratory, 4Navy Technology Center for Safety and Survivability, Chemistry Division, U.S. Naval Research Laboratory Trace explosive vapors of TNT and RDX collected on sorbent-filled thermal desorption tubes were analyzed using a programmed temperature desorption system coupled to GC with an electron capture detector. The instrumental analysis is combined with direct liquid deposition method to reduce sample variability and account for instrumentation drift and losses. Chemistry A Technique to Functionalize and Self-assemble Macroscopic Nanoparticle-ligand Monolayer Films onto Template-free Substrates Jake Fontana1, Christopher Spillmann1, Jawad Naciri1, Banahalli R. Ratna1 1Naval Research Laboratory A simple, robust and scalable technique to functionalize and self-assemble macroscopic nanoparticle-ligand monolayer films onto template-free substrates is described in this protocol. Immunology and Infection Locked Nucleic Acid Flow Cytometry-fluorescence in situ Hybridization (LNA flow-FISH): a Method for Bacterial Small RNA Detection Kelly L. Robertson1, Gary J. Vora1 1Center for Bio/Molecular Science and Engineering, Naval Research Laboratory A novel high-throughput method is described that enables the detection and relative quantitation of small RNA and mRNA expression from single bacterial cells using locked nucleic acid probes and flow cytometry-fluorescence in situ hybridization.
