Lawrence Livermore National Laboratory View Institution's Website 12 articles published in JoVE Immunology and Infection Cell-Free Scaled Production and Adjuvant Addition to a Recombinant Major Outer Membrane Protein from Chlamydia muridarum for Vaccine Development Sean F. Gilmore*1, Wei He*1, Angela C. Evans1, Delia F. Tifrea2, Sukumar Pal2, Brent Segelke1, Sandra K. G. Peters1, B. Dillon Vannest1, Nicholas O. Fischer1, Amy Rasley1, Luis M. de la Maza2, Matthew A. Coleman1,3 1Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory, 2Department of Pathology and Laboratory Medicine, University of California, 3School of Medicine, Radiation Oncology, University of California Davis This protocol describes using commercial, cell-free protein expression kits to produce membrane proteins supported in nanodisc that can be used as antigens in subunit vaccines. Neuroscience Chronic Implantation of Multiple Flexible Polymer Electrode Arrays Jason E Chung*1,2, Hannah R Joo*1,2, Clay N Smyth2, Jiang Lan Fan3, Charlotte Geaghan-Breiner2, Hexin Liang2, Daniel Fan Liu3, Demetris Roumis2, Supin Chen4,5, Kye Y Lee4, Jeanine A Pebbles4, Angela C Tooker4, Vanessa M Tolosa4,5, Loren M Frank2,6 1Medical Scientist Training Program and Neuroscience Graduate Program, University of California San Francisco, 2Kavli Institute for Fundamental Neuroscience, Center for Integrative Neuroscience, and Department of Physiology, University of California San Francisco, 3Bioengineering Graduate Program, University of California San Francisco, 4Center for Micro- and Nanotechnology, Lawrence Livermore National Laboratory, 5Neuralink Corp., 6Howard Hughes Medical Institute Described below is a method for implantation of multiple polymer electrode arrays across anatomically distant brain regions for chronic electrophysiological recording in freely moving rats. Preparation and surgical implantation are described in detail, with emphasis on design principles to guide adaptation of these methods for use in other species. Chemistry Electrochemical Roughening of Thin-Film Platinum Macro and Microelectrodes Anna N. Ivanovskaya*1, Anna M. Belle*1, Allison Yorita1, Fang Qian2, Supin Chen1, Angela Tooker1, Rose GarcÍa Lozada1, Dylan Dahlquist1, Vanessa Tolosa1 1Engineering Directorate, Lawrence Livermore National Laboratory, 2Physical and Life Science Directorate, Lawrence Livermore National Laboratory This protocol demonstrates a method for electrochemical roughening of thin-film platinum electrodes without preferential dissolution at grain boundaries. The electrochemical techniques of cyclic voltammetry and impedance spectroscopy are demonstrated to characterize these electrode surfaces. Chemistry Negative Additive Manufacturing of Complex Shaped Boron Carbides Ryan Lu1, Dorothy J. Miller1, Wyatt L. Du Frane1, Swetha Chandrasekaran1, Richard L. Landingham1, Marcus A. Worsley1, Joshua D. Kuntz1 1Lawrence Livermore National Laboratory A method called negative additive manufacturing is used to produce near fully dense complex shaped boron carbide parts of various length scales. This technique is possible via the formulation of a novel suspension involving resorcinol-formaldehyde as a unique gelling agent that leaves behind a homogenous carbon sintering aid after pyrolysis. Engineering Using Synchrotron Radiation Microtomography to Investigate Multi-scale Three-dimensional Microelectronic Packages Holly D. Carlton1, John W. Elmer1, Yan Li2, Mario Pacheco2, Deepak Goyal2, Dilworth Y. Parkinson3, Alastair A. MacDowell3 1Materials Engineering Division, Lawrence Livermore National Laboratory, 2Assembly Test and Technology Development Failure Analysis Labs, Intel Corporation, 3Advanced Light Source, Lawrence Berkeley National Laboratory For this study synchrotron radiation micro-tomography, a non-destructive three-dimensional imaging technique, is employed to investigate an entire microelectronic package with a cross-sectional area of 16 x 16 mm. Due to the synchrotron's high flux and brightness the sample was imaged in just 3 min with an 8.7 µm spatial resolution. Bioengineering A Microfluidic Platform for Precision Small-volume Sample Processing and Its Use to Size Separate Biological Particles with an Acoustic Microdevice Erika J. Fong1,2, Chao Huang1, Julie Hamilton1, William J. Benett1, Mihail Bora1, Alison Burklund1, Thomas R. Metz1, Maxim Shusteff1 1Materials Engineering Division, Lawrence Livermore National Laboratory, 2Department of Biomedical Engineering, Boston University This protocol describes a system architecture for performing automated small volume (0.15–1.5 ml) particle separations using a microfluidic device, and discusses methods to optimize acoustofluidic device performance and operation. Engineering Synthesis and Functionalization of 3D Nano-graphene Materials: Graphene Aerogels and Graphene Macro Assemblies Patrick G. Campbell1, Marcus A. Worsley1, Anna M. Hiszpanski1, Theodore F. Baumann1, Juergen Biener1 1Materials Science Division, Lawrence Livermore National Laboratory This video method describes the synthesis of high surface area, monolithic 3D graphene-based materials derived from polymer precursors as well as single layer graphene oxide. Chemistry Fabrication of Large-area Free-standing Ultrathin Polymer Films Michael Stadermann1, Salmaan H. Baxamusa1, Chantel Aracne-Ruddle1, Maverick Chea1, Shuaili Li1, Kelly Youngblood1, Tayyab Suratwala1 1Physical and Life Sciences Directorate, Lawrence Livermore National Laboratory We describe a method for the fabrication of large-area (up to 13 cm diameter) and ultrathin (as thin as 8 nm) polymer films. Instead of using a sacrificial interlayer to delaminate the film from its substrate, we use a self-limiting surface treatment suitable for arbitrarily large areas. Engineering Convergent Polishing: A Simple, Rapid, Full Aperture Polishing Process of High Quality Optical Flats & Spheres Tayyab Suratwala1, Rusty Steele1, Michael Feit1, Rebecca Dylla-Spears1, Richard Desjardin1, Dan Mason1, Lana Wong1, Paul Geraghty1, Phil Miller1, Nan Shen1 1Lasers, Optics, & Targets for the National Ignition Facility, Lawrence Livermore National Laboratory A novel optical polishing process, called “Convergent Polishing”, which enables faster, lower cost polishing, is described. Unlike conventional polishing processes, Convergent Polishing allows a glass workpiece to be polished in a single iteration and with high surface quality to its final surface figure without requiring changes to polishing parameters. Bioengineering The Portable Chemical Sterilizer (PCS), D-FENS, and D-FEND ALL: Novel Chlorine Dioxide Decontamination Technologies for the Military Christopher J. Doona1, Florence E. Feeherry1, Peter Setlow2, Alexander J. Malkin3, Terrence J. Leighton4 1United States Army-Natick Soldier RD&E Center, Warfighter Directorate, 2Department of Molecular Biology and Biophysics, University of Connecticut Health Center, 3Lawrence Livermore National Laboratory, 4 The Portable Chemical Sterilizer (PCS) is a revolutionary, energy-independent, almost waterless sterilization technology for Army medical units. The PCS generates chlorine dioxide from dry reagents mixed with water on-site, at-will, and at point-of-use (PoU) in a plastic suitcase. The Disinfectant-sprayer for Foods and ENvironmentally-friendly Sanitation (D-FENS) and the Disinfectant for ENvironmentally-friendly Decontamination, All-purpose (D-FEND ALL) produce aqueous chlorine dioxide in a collapsible spray bottle and other potential embodiments. These versatile decontamination technologies kill microbes in myriad diverse Dual-use applications for military and civilian consumers. Bioengineering Insertion of Flexible Neural Probes Using Rigid Stiffeners Attached with Biodissolvable Adhesive Sarah H. Felix1, Kedar G. Shah1, Vanessa M. Tolosa1, Heeral J. Sheth1, Angela C. Tooker1, Terri L. Delima1, Shantanu P. Jadhav2, Loren M. Frank2, Satinderpall S. Pannu1 1Materials Engineering Division, Lawrence Livermore National Laboratory, 2UCSF Center for Integrative Neuroscience and the Department of Physiology, University of California, San Francisco Insertion of flexible neural microelectrode probes is enabled by attaching probes to rigid stiffeners with polyethylene glycol (PEG). A unique assembly process ensures uniform and repeatable attachment. After insertion into tissue, the PEG dissolves and the stiffener is extracted. An in vitro test method evaluates the technique in agarose gel. Bioengineering Planar and Three-Dimensional Printing of Conductive Inks Bok Yeop Ahn1, Steven B. Walker1, Scott C. Slimmer1, Analisa Russo1, Ashley Gupta1, Steve Kranz1, Eric B. Duoss1,2, Thomas F. Malkowski1,3, Jennifer A. Lewis1 1Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, 2Center for Micro- and Nanotechnology, Lawrence Livermore National Laboratory, 3Presently at the Interdisciplinary Center for Wide Band-gap Semiconductors, University Of California Santa Barbara Planar and three-dimensional printing of conductive metallic inks is described. Our approach provides new avenues for fabricating printed electronic, optoelectronic, and biomedical devices in unusual layouts at the microscale.
