City University of New York Graduate Center 9 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. 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. Cancer Research Surface-enhanced Resonance Raman Scattering Nanoprobe Ratiometry for Detecting Microscopic Ovarian Cancer via Folate Receptor Targeting Chrysafis Andreou1, Anton Oseledchyk1, Fay Nicolson1, Naxhije Berisha1,2, Suchetan Pal1, Moritz F. Kircher1,3,4,5,6,7 1Department of Radiology, Memorial Sloan Kettering Cancer Center, 2Department of Chemistry, The Graduate Center of the City University of New York, 3Molecular Pharmacology Program, Memorial Sloan Kettering Cancer Center, 4Center for Molecular Imaging and Nanotechnology (CMINT), Memorial Sloan Kettering Cancer Center, 5Gerstner Sloan Kettering Graduate School of Biomedical Sciences, 6Department of Radiology, Weill Cornell Medical College of Cornell University, 7Dana-Farber Cancer Institute and Harvard Medical Center Ovarian cancer forms metastases throughout the peritoneal cavity. Here, we present a protocol to make and use folate-receptor targeted surface-enhanced resonance Raman scattering nanoprobes that reveal these lesions with high specificity via ratiometric imaging. The nanoprobes are administered intraperitoneally to living mice, and the derived images correlate well with histology. Genetics Characterizing Histone Post-translational Modification Alterations in Yeast Neurodegenerative Proteinopathy Models Seth A. Bennett1,2, Samantha N. Cobos1,3, Marcella Meykler1, Michel Fallah1, Navin Rana1, Karen Chen1, Mariana P. Torrente1,4 1Chemistry Department, Brooklyn College, 2Ph.D. Program in Biochemistry, Graduate Center of the City University of New York, 3Ph.D. Program in Chemistry, Graduate Center of the City University of New York, 4Ph.D. Programs in Chemistry, Biochemistry, and Biology, Graduate Center of the City University of New York This protocol outlines experimental procedures to characterize genome-wide changes in the levels of histone post-translational modifications (PTM) occurring in connection with the overexpression of proteins associated with ALS and Parkinson's disease in Saccharomyces cerevisiae models. After SDS-PAGE separation, individual histone PTM levels are detected with modification-specific antibodies via Western blotting. Chemistry Synthesis and Bioconjugation of Thiol-Reactive Reagents for the Creation of Site-Selectively Modified Immunoconjugates Maria Davydova1, Guillaume Dewaele Le Roi1,2, Pierre Adumeau1, Brian M. Zeglis1,2,3,4 1Department of Chemistry, Hunter College of the City University of New York, 2Ph.D. Program in Chemistry, Graduate Center of the City University of New York, 3Department of Radiology, Memorial Sloan Kettering Cancer Center, 4Department of Radiology, Weill Cornell Medical College In this protocol, we will describe the synthesis of PODS, a phenyoxadiazolyl methyl sulfone-based reagent for the site-selective attachment of cargos to the thiols of biomolecules, particularly antibodies. In addition, we will describe the synthesis and characterization of a PODS-bearing bifunctional chelator and its conjugation to a model antibody. Bioengineering A Tripeptide-Stabilized Nanoemulsion of Oleic Acid Sylwia A. Dragulska1, Marek T. Wlodarczyk1,2, Mina Poursharifi1,3, John A. Martignetti4,5,6, Aneta J. Mieszawska1,2,3 1Department of Chemistry, Brooklyn College, The City University of New York, 2Ph.D. Program in Chemistry, The Graduate Center of The City University of New York, 3Ph.D. Program in Biochemistry, The Graduate Center of The City University of New York, 4Department of Genetics and Genomic Sciences, Icahn School of Medicine at Mount Sinai, 5Women's Health Research Institute, Icahn School of Medicine at Mount Sinai, 6Laboratory for Translational Research, Western Connecticut Health Network This protocol describes an efficient method to synthesize a nanoemulsion of an oleic acids-platinum(II) conjugate stabilized with a lysine-tyrosine-phenylalanine (KYF) tripeptide. The nanoemulsion forms under mild synthetic conditions via self-assembly of the KYF and the conjugate. Chemistry Pretargeted Radioimmunotherapy Based on the Inverse Electron Demand Diels-Alder Reaction Rosemery Membreno1,2, Brendon E. Cook1,2,3, Brian M. Zeglis1,2,3,4 1Department of Chemistry, Hunter College of the City University of New York, 2Ph.D. Program in Chemistry, Graduate Center of the City University of New York, 3Department of Radiology, Memorial Sloan Kettering Cancer Center, 4Department of Radiology, Weill Cornell Medical College This protocol describes the synthesis and characterization of a trans-cyclooctene (TCO)-modified antibody and a 177Lu-labeled tetrazine (Tz) radioligand for pretargeted radioimmunotherapy (PRIT). In addition, it details the use of these two constructs for in vivo biodistribution and longitudinal therapy studies in a murine model of colorectal cancer. Bioengineering A Novel Technique for Generating and Observing Chemiluminescence in a Biological Setting Gabriel E. Büchel1,2, Brandon Carney1,3, Jun Tang1, Brian M. Zeglis1,3, Jörg Eppinger2, Thomas Reiner1,4 1Department of Radiology, Memorial Sloan Kettering Cancer Center, 2KAUST Catalysis Center, King Abdullah University of Science and Technology, 3Department of Chemistry, Hunter College, and PhD Program in Chemistry, Graduate Center of City University of New York, 4Department of Radiology, Weill Cornell Medical College This protocol describes a new intraoperative imaging technique that uses a ruthenium complex as a source of chemiluminescent light emission, thereby producing high signal-to-noise ratios during in vivo imaging. Intraoperative imaging is an expanding field that could revolutionize the way that surgical procedures are performed. Behavior Ultrasound Images of the Tongue: A Tutorial for Assessment and Remediation of Speech Sound Errors Jonathan L. Preston1,2, Tara McAllister Byun3, Suzanne E. Boyce2,4, Sarah Hamilton4, Mark Tiede2, Emily Phillips2, Ahmed Rivera-Campos4, Douglas H. Whalen2,5,6 1Department of Communication Sciences and Disorders, Syracuse University, 2Haskins Laboratories, 3Department of Communicative Sciences and Disorders, New York University, 4Department of Communication Sciences and Disorders, University of Cincinnati, 5Program in Speech-Language-Hearing Sciences, City University of New York Graduate Center, 6Department of Linguistics, Yale University Ultrasound imaging can be used to display the shape and movements of the tongue in real time during speech. The images can be used to determine the nature of speech sound errors. Visual feedback of the tongue can be used to facilitate improvements in speech sound production in clinical populations.
