Columbia University Medical Center 12 articles published in JoVE Bioengineering An Ex Vivo Porcine Model for Hydrodynamic Testing of Experimental Aortic Valve Procedures and Novel Medical Devices V. Reed LaSala1, Halil Beqaj2, Mingze Sun1, Sabrina Castagnini3, Senay Ustunel1, Elizabeth Cordoves2, Kavya Rajesh2, Sophia Jackman2, David Kalfa1,4 1Department of Surgery, Columbia University Medical Center, 2College of Physicians and Surgeons, Columbia University, 3School of Medicine and Surgery, University of Bologna, 4Division of Cardiac, Thoracic, and Vascular Surgery, Section of Pediatric and Congenital Cardiac Surgery, New York-Presbyterian Morgan Stanley Children’s Hospital, Columbia University Medical Center We present a method for mounting a porcine aortic valve on a pulse duplicator to test its hydrodynamic properties. This method can be used to determine the change in hydrodynamics after the application of an experimental procedure or novel medical device prior to use in a large animal model. Biology Assessment of Open Probability of the Mitochondrial Permeability Transition Pore in the Setting of Coenzyme Q Excess Keren K. Griffiths1, Aili Wang1, Richard J. Levy1 1Department of Anesthesiology, Columbia University Medical Center This method exploits the contribution of the mitochondrial permeability transition pore to low-conductance proton leak to determine the voltage threshold for pore opening in neonatal fragile X syndrome mice with increased cardiomyocyte mitochondrial coenzyme Q content compared to wildtype control. Developmental Biology A Protocol for Immunohistochemistry and RNA In-situ Distribution within Early Drosophila Embryo Wei Zhang*1, Xinjuan Lei*1, Xin Zhou*2,3, Boling He1, Liqin Xiao1, Huimin Yue1, Shulin Wang1, Yuting Sun1, Yajun Wu1, Liyang Wang1,4, George Ghartey-Kwansah1, Odell D. Jones5, Joseph L. Bryant6, MengMeng Xu7, Jianjie Ma3, Xuehon Xu1 1National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China/CGDB, Shaanxi Normal University College of Life Sciences, 2 Here, we describe a protocol for detection and localization of Drosophila embryo protein and RNA from collection to pre-embedding and embedding, immunostaining, and mRNA in situ hybridization. Medicine Modified Technique for the Use of Neonatal Murine Hearts in the Langendorff Preparation Matthew B. Barajas1, Richard J. Levy2 1Department of Anesthesiology, Vanderbilt University Medical Center, 2Department of Anesthesiology, Columbia University Medical Center The present protocol describes aortic cannulation and retrograde perfusion of the ex-vivo neonatal murine heart. A two-person strategy, using a dissecting microscope and a blunted small gauge needle, permits reliable cannulation. Quantification of longitudinal contractile tension is achieved using a force transducer connected to the apex of the left ventricle. Bioengineering A Large Animal Model for Pulmonary Hypertension and Right Ventricular Failure: Left Pulmonary Artery Ligation and Progressive Main Pulmonary Artery Banding in Sheep Rei Ukita1, John W. Stokes1, W. Kelly Wu1, Jennifer Talackine1, Nancy Cardwell1, Yatrik Patel1, Clayne Benson5, Caitlin T. Demarest1, Erika B. Rosenzweig3, Keith Cook2, Emily J. Tsai4, Matthew Bacchetta1,6 1Department of Thoracic Surgery, Vanderbilt University Medical Center, 2Department of Biomedical Engineering, Carnegie Mellon University, 3Department of Pediatrics, Columbia University Medical Center, Columbia University Irving Medical Center, 4Department of Medicine, Columbia University Medical Center, Columbia University Irving Medical Center, 5Department of Anesthesiology, Vanderbilt University Medical Center, 6Department of Biomedical Engineering, Vanderbilt University This manuscript describes the surgical technique and experimental approach to develop severe right ventricular pressure overload to model their adaptive and maladaptive phenotypes. Genetics Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant Harpegnathos saltator Kayli Sieber1, Maya Saar1, Comzit Opachaloemphan2, Matthew Gallitto3, Huan Yang2, Hua Yan1,4 1Department of Biology, University of Florida, 2Department of Biochemistry and Molecular Pharmacology, New York University School of Medicine, 3Department of Radiation Oncology, Columbia University Medical Center, 4Center for Smell and Taste, University of Florida Many characteristics of insect eusociality rely on within-colony communication and division of labor. Genetic manipulation of key regulatory genes in ant embryos via microinjection and CRISPR-mediated mutagenesis provides insights into the nature of altruistic behavior in eusocial insects. Medicine Ex Vivo Corneal Organ Culture Model for Wound Healing Studies Nileyma Castro*1, Stephanie R. Gillespie*2, Audrey M. Bernstein1 1Department of Ophthalmology, SUNY Upstate Medical University, 2Department of Dermatology, Columbia University Medical Center A protocol for an ex vivo corneal organ culture model useful for wound healing studies is described. This model system can be used to assess the effects of agents to promote regenerative healing or drug toxicity in an organized 3D multicellular environment. Immunology and Infection Assessing Cellular Stress and Inflammation in Discrete Oxytocin-secreting Brain Nuclei in the Neonatal Rat Before and After First Colostrum Feeding Benjamin Y. Klein1, Hadassah Tamir1,2,3, Muhammad Anwar1, Robert J. Ludwig1, Jasmine H. Kaidbey1, Sara B. Glickstein4, Martha G. Welch1,2 1Department of Psychiatry, Columbia University, 2Department of Pathology & Cell Biology, Columbia University Medical Center, 3Department of Psychiatry, Division of Molecular Imaging and Neuropathology, New York State Psychiatric Institute, 4EB Sciences Here, we present a protocol to isolate brain nuclei in the neonatal rat brain in conjunction with first colostrum feeding. This technique allows the study of nutrient insufficiency stress in the brain as modulated by enterocyte signaling. Behavior A Within-subjects Experimental Protocol to Assess the Effects of Social Input on Infant EEG Ashley M. St. John1, Katie Kao1, Meia Chita-Tegmark1, Jacqueline Liederman1, Philip G. Grieve2, Amanda R. Tarullo1 1Department of Psychological and Brain Sciences, Boston University, 2Department of Pediatrics, Columbia University Medical Center This novel protocol is designed to assess the neural bases of social interaction in infants. The paradigm is designed to tease apart how various social inputs such as language, joint attention, and face-to-face interaction relate to infant neural activation. Infant EEG power is recorded during both social and nonsocial conditions. Neuroscience Stereotaxic Infusion of Oligomeric Amyloid-beta into the Mouse Hippocampus Ying Y. Jean1, Jimena Baleriola2, Mauro Fà2, Ulrich Hengst1,2, Carol M. Troy1,2,3 1Department of Pathology and Cell Biology, Columbia University Medical Center, 2The Taub Institute for Research on Alzheimer's Disease and the Aging Brain, Columbia University Medical Center, 3Department of Neurology, Columbia University Medical Center Here, we present a protocol for direct stereotaxic brain infusion of amyloid-beta. This methodology provides an alternative in vivo mouse model to address the short-term effects of amyloid-beta on brain neurons. Neuroscience Investigating the Function of Deep Cortical and Subcortical Structures Using Stereotactic Electroencephalography: Lessons from the Anterior Cingulate Cortex Robert A. McGovern1,3, Tarini Ratneswaren4, Elliot H. Smith1,3, Jennifer F. Russo3, Amy C. Jongeling2,3, Lisa M. Bateman2,3, Catherine A. Schevon2,3, Neil A. Feldstein1,3, Guy M. McKhann, II1,3, Sameer Sheth1,3 1Department of Neurosurgery, Columbia University Medical Center, New York Presbyterian Hospital, 2Department of Neurology, Columbia University Medical Center, New York Presbyterian Hospital, 3Columbia University Medical Center, New York Presbyterian Hospital, 4 Stereotactic Electroencephalography (SEEG) is an operative technique used in epilepsy surgery to help localize seizure foci. It also affords a unique opportunity to investigate brain function. Here we describe how SEEG can be used to investigate cognitive processes in human subjects. Immunology and Infection In vivo Imaging Method to Distinguish Acute and Chronic Inflammation Jen-Chieh Tseng1, Andrew L. Kung2 1Lurie Family Imaging Center, Dana-Farber Cancer Institute, Harvard Medical School, 2Division of Pediatric Hematology/Oncology/Stem Cell Transplantation, Columbia University Medical Center We describe a non-invasive imaging method for distinguishing inflammatory stages. Systemic delivery of luminol reveals areas of acute inflammation dependent upon MPO activity in neutrophils. In contrast, injection of lucigenin allows for visualization of chronic inflammation dependent upon Phox activity in macrophages.
