Technion View Institution's Website 6 articles published in JoVE Bioengineering Stepwise Cell Seeding on Tessellated Scaffolds to Study Sprouting Blood Vessels Ariel A. Szklanny1, Dylan B. Neale2, Joerg Lahann2, Shulamit Levenberg1 1Faculty of Biomedical Engineering, Technion, 2Department of Chemical Engineering and Biointerfaces Institute, University of Michigan, Ann Arbor Engineered tissues heavily rely on proper vascular networks to provide vital nutrients and gases and remove metabolic waste. In this work, a stepwise seeding protocol of endothelial cells and support cells creates highly organized vascular networks in a high-throughput platform for studying developing vessel behavior in a controlled 3D environment. Biology Live Imaging of Apoptotic Cell Clearance during Drosophila Embryogenesis Boris Shklyar1, Jeny Shklover1, Estee Kurant1 1Department of Anatomy and Cell Biology and the Rappaport Institute for Research in the Medical Sciences, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology Here we describe an effective method for studying dynamics of apoptotic cell clearance in vivo. This method employs live Drosophila embryos as a powerful model for monitoring phagocytosis of apoptotic cells using specific labeling of apoptotic cells and phagocytes. Biology Preparation of Cell-lines for Conditional Knockdown of Gene Expression and Measurement of the Knockdown Effects on E4orf4-Induced Cell Death Anna Brestovitsky1, Rakefet Sharf1, Tamar Kleinberger1 1Department of Molecular Microbiology, Faculty of Medicine, Technion - Israel Institute of Technology Contribution of the ACF chromatin remodeling factor to E4orf4-induced cell death was measured. The protocol includes selection of cell clones in which doxycycline treatment induces conditional knockdown of the ACF subunits Acf1 and SNF2h, and use of the DAPI assay to measure E4orf4-induced cell death in the inducible cell lines. Neuroscience Visualization of Proprioceptors in Drosophila Larvae and Pupae Naomi Halachmi1, Atalya Nachman1, Adi Salzberg1 1Department of Genetics and the Rappaport Institute for Research in the Medical Sciences, Rappaport Faculty of Medicine, Technion-Israel Institute of Technology A method to immunostain and visualize chordotonal organs in larvae and pupae of Drosophila melanogaster is described. Immunology and Infection In vitro tRNA Methylation Assay with the Entamoeba histolytica DNA and tRNA Methyltransferase Dnmt2 (Ehmeth) Enzyme Ayala Tovy1, Benjamin Hofmann2, Mark Helm2, Serge Ankri1 1Faculty of Medicine, Rappaport Institute, Technion - Israel Institute of Technology, 2The Pharmacy and Biochemistry Institute, Johannes Gutenberg University This protocol describes the preparation of a synthetic tRNA substrate for the Entamoeba histolytica DNA/tRNA methyltransferase 2 (Dnmt2) homolog Ehmeth and the measure of its methyltransferase activity. This experimental approach can be used for investigating the activity of other Dnmt2 proteins. Biology Electron Spin Resonance Micro-imaging of Live Species for Oxygen Mapping Revital Halevy1, Lazar Shtirberg1, Michael Shklyar1, Aharon Blank1 1Schulich Faculty of Chemistry, The Technion, Israel Institute of Technology This protocol describes a method for micron-scale three-dimensional imaging of oxygen concentration in the immediate environment of live cells by electron spin resonance microscopy.