The City University of New York View Institution's Website 13 articles published in JoVE Biology Sample Preparation for Rapid Lipid Analysis in Drosophila Brain Using Matrix-Assisted Laser Desorption/Ionization Mass Spectrometry Imaging Yuki X. Chen*1,5,6, Kelly Veerasammy*1,2, Jun Yin*3, Tenzin Choetso1,4, Tiffany Zhong7, Muniyat A. Choudhury1,4,5, Cory Weng1,4,5, Ethan Xu8, Mayan A. Hein9,10, Rinat Abzalimov2, Ye He1 1Advanced Science Research Center, Neuroscience Initiative, the City University of New York, Graduate Center New York, 2Advanced Science Research Center, Structural Biology Initiative, the City University of New York, Graduate Center, 3National Institute of Neurological Disorders and Stroke, National Institutes of Health, 4The City College of New York, CUNY, 5Macaulay Honors College, CUNY, 6Graduate School of Public Health and Health Policy, The City University of New York, 7Princeton University, 8Ardrey Kell High School, 9The Borough of Manhattan Community College, CUNY, 10Gallatin School of Individualized Study, New York University The aim of this protocol is to provide detailed guidance on the proper sample preparation for lipid and metabolite analysis in small tissues, such as the Drosophila brain, using matrix-assisted laser desorption/ionization (MALDI) mass spectrometry imaging. Neuroscience A Plate-Based Assay for the Measurement of Endogenous Monoamine Release in Acute Brain Slices Jose A. Pino*1, Nora Awadallah*2, Alessandra M. Norris3, Gonzalo E. Torres2 1Department of Medicine, School of Medicine, Universidad de Atacama, 2Department of Molecular, Cellular, and Biomedical Sciences, City University of New York School of Medicine at City College, 3Department of Pharmacology and Therapeutics, University of Florida College of Medicine This method introduces a simple technique for the detection of endogenous monoamine release using acute brain slices. The setup uses a 48-well plate containing a tissue holder for monoamine release. Released monoamine is analyzed by HPLC coupled with electrochemical detection. Additionally, this technique provides a screening method for drug discovery. Biology Sample Preparation for Metabolic Profiling using MALDI Mass Spectrometry Imaging Kelly Veerasammy*1,2, Yuki X. Chen*1,2, Sami Sauma*1, Mathilde Pruvost1, David K. Dansu1, Tenzin Choetso1,2, Tiffany Zhong3, Damien Marechal1, Patrizia Casaccia1, Rinat Abzalimov4,5, Ye He1,5 1The Graduate Center - Advanced Science Research Center, Neuroscience Initiative, The City University of New York, 2The City College of New York, CUNY, 3The Bronx High School of Science, 4The Graduate Center - Advanced Science Research Center, Structural Biology Initiative, The City University of New York, 5The Graduate Center - Advanced Science Research Center, MALDI MS Imaging Joint Core Facility, The City University of New York The goal of this protocol is to provide detailed guidance on the sample preparation when planning for experiments using MALDI MSI to maximize metabolic and molecular detection in biological samples. Environment Simulating Impacts of Ice Storms on Forest Ecosystems John L. Campbell1, Lindsey E. Rustad1, Charles T. Driscoll2, Ian Halm3, Timothy J. Fahey4, Habibollah Fakhraei5, Peter M. Groffman6,7, Gary J. Hawley8, Wendy Leuenberger9, Paul G. Schaberg10 1Northern Research Station, U.S. Forest Service, Durham, NH, 2Department of Civil and Environmental Engineering, Syracuse University, 3Northern Research Station, U.S. Forest Service, North Woodstock, NH, 4Department of Natural Resources, Cornell University, 5Department of Civil and Environmental Engineering, Southern Illinois University, 6Advanced Science Research Center at the Graduate Center, City University of New York, 7Cary Institute of Ecosystem Studies, 8Rubenstein School of Environment and Natural Resources, University of Vermont, 9Department of Forestry and Natural Resources, University of Kentucky, 10Northern Research Station, U.S. Forest Service, Burlington, VT Ice storms are important weather events that are challenging to study because of difficulties in predicting their occurrence. Here, we describe a novel method for simulating ice storms that involves spraying water over a forest canopy during sub-freezing conditions. Biology Visualizing and Tracking Endogenous mRNAs in Live Drosophila melanogaster Egg Chambers Irina E. Catrina1, Livia V. Bayer1,2, Omar S. Omar1,2, Diana P. Bratu1,2 1Biological Sciences Department Hunter College, City University of New York, 2Program in Molecular, Cellular, and Developmental Biology, Graduate Center, City University of New York Here, we present a protocol for the visualization, detection, analysis and tracking of endogenous mRNA trafficking in live Drosophila melanogaster egg chamber using molecular beacons, spinning disc confocal microscopy, and open-source analysis software. Genetics Manipulation of Ploidy in Caenorhabditis elegans Erlyana K. Clarke1, Katherine A. Rivera Gomez1,2, Zaki Mustachi1, Mikaela C. Murph1,3, Mara Schvarzstein1,2,3 1Brooklyn College, Biology Department, City University of New York, 2The Graduate Center, Biology Department, City University of New York, 3Advanced Science Research Center, City University of New York This method allows for the generation of tetraploid and triploid Caenorhabditis nematodes from any diploid strain. Polyploid strains generated by this method have been used to study chromosome interactions in meiotic prophase, and this method is useful for examining important basic questions in cell, developmental, evolutionary, and cancer biology. Neuroscience Simultaneous Detection of c-Fos Activation from Mesolimbic and Mesocortical Dopamine Reward Sites Following Naive Sugar and Fat Ingestion in Rats Julie A. D. Dela Cruz*1, Tricia Coke*2, Richard J. Bodnar2,3 1Behavioral and Cognitive Neuroscience Cluster, Psychology Doctoral Program, The Graduate Center, CUNY, New York, NY, 2Department of Psychology, Queens College, CUNY, Flushing, NY, 3Behavioral and Cognitive Neuroscience Cluster, Psychology Doctoral Program, The Graduate Center, CUNY, Flushing, NY The goal of this study is to identify reward-related distributed brain networks by delineating a reliable immunohistological technique using cellular c-fos activation to measure simultaneous changes in dopamine pathways and terminal sites after novel ingestion of fat and sugar in rats. Medicine A "Patient-Like" Orthotopic Syngeneic Mouse Model of Hepatocellular Carcinoma Metastasis Dibash K. Das1,2, Victoria Durojaiye1, Adeodat Ilboudo1, Michelle K. Naidoo1, Olorunseun Ogunwobi1,2 1Department of Biological Sciences, Hunter College of the City University of New York, 2Departments of Biology and Biochemistry, The Graduate Center of the City University of New York The metastatic spread of cancer is the major cause of cancer-related deaths. We provide an in-depth description of our survival surgery methodology for establishing a “patient-like” orthotopic syngeneic mouse model system for studying the mechanisms of metastasis in solid organ tumors. Medicine Isolation and Propagation of Circulating Tumor Cells from a Mouse Cancer Model Dibash K. Das1,2, Michelle K. Naidoo1, Adeodat Ilboudo1, Pascal DuBois1, Victoria Durojaiye1, Chen Liu3, Olorunseun O. Ogunwobi1,2 1Department of Biological Sciences, Hunter College of The City University of New York, 2Departments of Biology and Biochemistry, The Graduate Center of The City University of New York, 3Department of Pathology, Immunology, and Laboratory Medicine, University of Florida Circulating tumor cells (CTCs) have been shown to play an important role in tumor metastasis. Here, a method for the isolation and propagation of CTCs from the whole blood of a syngeneic mouse tumor model of hepatocellular carcinoma (HCC) metastasis is described. Chemistry Diffuse Reflectance Infrared Spectroscopic Identification of Dispersant/Particle Bonding Mechanisms in Functional Inks L. Jay Deiner1, Elaheh Farjami1 1Department of Chemistry, New York City College of Technology, City University of New York (CUNY) Formulation of stable, functional inks is critical to expanding the applications of additive manufacturing. In turn, knowledge of the mechanisms of dispersant/particle bonding is required for effective ink formulation. Diffuse Reflectance Fourier Transform Infrared Spectroscopy (DRIFTS) is presented as a simple, inexpensive way to gain insight into these mechanisms. Immunology and Infection Derivation of T Cells In Vitro from Mouse Embryonic Stem Cells Martina Kučerová-Levisohn1, Jordana Lovett1, Armin Lahiji1, Roxanne Holmes2, Juan Carlos Zúñiga-Pflücker2, Benjamin D. Ortiz1 1Department of Biological Sciences, Hunter College and Graduate Center, City University of New York, 2Sunnybrook Research Institute, Department of Immunology, University of Toronto Mouse embryonic stem cells can be differentiated to T cells in vitro using the OP9-DL1 co-culture system. Success in this procedure requires careful attention to reagent/cell maintenance, and key technique sensitive steps. Here we discuss these critical parameters and provide a detailed protocol to encourage adoption of this technology. Neuroscience A Method to Make a Craniotomy on the Ventral Skull of Neonate Rodents Adrián Rodríguez-Contreras1, Lingyan Shi2, Bingmei M. Fu2 1Department of Biology, The City University of New York, City College, 2Department of Biomedical Engineering, The City University of New York, City College A surgical method is described to expose the ventral skull in neonate rats. Using this approach it is possible to open a craniotomy to perform acute electrophysiology and two-photon microscopy experiments in the brainstem of anesthetized pups. Immunology and Infection An Introduction to Parasitic Wasps of Drosophila and the Antiparasite Immune Response Chiyedza Small*1,2, Indira Paddibhatla*1,2, Roma Rajwani1, Shubha Govind1,2 1Biology Department, The City College of New York, CUNY, 2The Graduate Center, The City University of New York Parasitoid (parasitic) wasps constitute a major class of natural enemies of many insects including Drosophila melanogaster. We will introduce the techniques to propagate these parasites in Drosophila spp. and demonstrate how to analyze their effects on immune tissues of Drosophila larvae.