Silk Fibroin Film Culture System for In vitro Analysis and Biomaterial Design Brian D. Lawrence1, Zhi Pan2, 1, Michael D. Weber1, David L. Kaplan3, Mark I. Rosenblatt11Margaret M. Dyson Vision Research Institute, Weill Cornell Medical College , 21Margaret M. Dyson Vision Research Institute, Weill Cornell Medical College , 3Department of Biomedical Engineering, Tufts University Silk films are a novel class of biomaterials readily customizable for an array of biomedical applications. The presented silk film culture system is highly adaptable to a variety of in vitro analyses. This system represents a biomaterial design platform offering in vitro optimization before direct translation to in vivo models. Induction of Myocardial Infarction in Adult Zebrafish using Cryoinjury Fabian Chablais, Anna JaźwińskaDepartment of Biology, Unit of Zoology, University of Fribourg, Fribourg, Switzerland Zebrafish represents a valuable model to study the mechanisms of heart regeneration in vertebrates. Here, we present a protocol for induction of a heart infarct in adult zebrafish using cryoinjury. This method results in massive cell death within 20% of the ventricular wall, similar to that observed in mammalian infarcts. Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography Todd A. Sasser1, Sarah E. Chapman2, Shengting Li1, Caroline Hudson2, Sean P. Orton1, Justin M. Diener3, Seth T. Gammon1, Carlos Correcher4, W. Matthew Leevy21Carestream Molecular Imaging , 2Department of Chemistry and Biochemistry, University of Notre Dame , 3Freimann Life Science Center, University of Notre Dame , 4Research and Development, Oncovision, GEM-Imaging S.A. Fat content analysis is routinely conducted in studies utilizing murine obesity models. Emerging methods in small animal CT imaging and analysis are providing for longitudinal detail rich fat content analysis. Here we detail step by step procedures for performing small animal CT imaging, analysis, and visualization. Stereotactic Radiosurgery for Gynecologic Cancer Charles Kunos1, James M. Brindle1, Robert Debernardo21Department of Radiation Oncology, University Hospitals Case Medical Center and Case Western Reserve University School of Medicine, 2Department of Obstetrics and Gynecology, Division of Gynecologic Oncology, University Hospitals Case Medical Center and Case Western Reserve University School of Medicine Stereotactic body radiotherapy (SBRT) involves image-guided, ablative radiation delivered to cancer targets refractory to chemotherapy or to conventional radiation treatment. The robotic-armed Cyberknife SBRT system, using sophisticated target localization, delivers hypofractionated radiation doses capable of sterilizing cancer targets. This article will consider new therapeutic roles of SBRT for gynecological cancers. Establishment of Microbial Eukaryotic Enrichment Cultures from a Chemically Stratified Antarctic Lake and Assessment of Carbon Fixation Potential Jenna M. Dolhi, Nicholas Ketchum, Rachael M. Morgan-KissDepartment of Microbiology, Miami University Microbial eukaryotes are both a source of photosynthetically-derived carbon and top predatory species in permanently ice-covered Antarctic lakes. This report describes an enrichment culture approach to isolate metabolically versatile microbial eukaryotes from the Antarctic lake, Lake Bonney, and assesses inorganic carbon fixation potential using a radioisotope assay for Ribulose-1,5-bisphophate carboxylase oxygenase (RubisCO) activity. Retrograde Loading of Nerves, Tracts, and Spinal Roots with Fluorescent Dyes Dvir Blivis, Michael J. O'DonovanDevelopmental Neurobiology Section, National Institute of Neurological Disorders and Stroke, National Institutes of Health We describe a simple and low cost technique for introducing high concentration of fluorescent and calcium-sensitive dyes into neurons or any neuronal tract using a polyethylene suction pipette. A Primary Neuron Culture System For The Study Of Herpes Simplex Virus Latency And Reactivation Mariko Kobayashi1, Ju-Youn Kim1, Vladimir Camarena2, Pamela C. Roehm3, Moses V. Chao4, 2, 5, 6, 7, Angus C. Wilson1, Ian Mohr11Department of Microbiology, New York University School of Medicine, 2Molecular Neurobiology Program, Skirball Institute for Biomolecular Medicine, New York University School of Medicine, 3Department of Otolaryngology, New York University School of Medicine, 4Department of Cell Biology, New York University School of Medicine, 5Department of Physiology and Neuroscience, New York University School of Medicine, 6Department of Psychiatry, New York University School of Medicine, 7Center for Neural Science, New York University School of Medicine The protocol describes an efficient and reproducible model system to study herpes simplex virus type 1 (HSV-1) latency and reactivation. The assay employs homogenous sympathetic neuron cultures and allows for the molecular dissection of virus-neuron interactions using a variety of tools including RNA interference and expression of recombinant proteins.
