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Columbia University Medical Center

15 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

In vivo Imaging Method to Distinguish Acute and Chronic Inflammation
Jen-Chieh Tseng 1, Andrew L. Kung 2
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.

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JoVE Journal

Static Adhesion Assay for the Study of Integrin Activation in T Lymphocytes
Marianne Strazza 1, Inbar Azoulay-Alfaguter 1, Ariel Pedoeem 1, Adam Mor 1,2
1Department of Medicine, New York University School of Medicine, 2Departments of Pathology, New York University School of Medicine

Static adhesion assay is a powerful tool that can be used to model the interactions between T lymphocytes and other cell types. Interactions are generated by injecting labeled T cells into wells coated with adhesion molecules, while a plate reader is used to quantify the number of adherent cells following serial washes.

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Neuroscience

Detection of Axonally Localized mRNAs in Brain Sections Using High-Resolution In Situ Hybridization
Jimena Baleriola 1, Ying Jean 1, Carol Troy 1, Ulrich Hengst 1
1College of Physicians and Surgeons, Columbia University

RNA in situ hybridization (ISH) enables the visualization of RNAs in cells and tissues. Here we show how combination of RNAscope ISH with immunohistochemistry or histological dyes can be successfully used to detect mRNAs localized to axons in sections of mouse and human brains.

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Neuroscience

Stereotaxic Infusion of Oligomeric Amyloid-beta into the Mouse Hippocampus
Ying Y. Jean 1, Jimena Baleriola 2, Mauro Fà 2, Ulrich Hengst 1,2, Carol M. Troy 1,2,3
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.

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Immunology and Infection

Assay of Adhesion Under Shear Stress for the Study of T Lymphocyte-Adhesion Molecule Interactions
Marianne Strazza 1, Inbar Azoulay-Alfaguter 1, Michael Peled 1, Adam Mor 1,2
1Department of Medicine, New York University School of Medicine, 2Department of Pathology, New York University School of Medicine

This flow adhesion assay provides a simple, high impact model of T cell-epithelial cell interactions. A syringe pump is used to generate shear stress, and confocal microscopy captures images for quantification. The goal of these studies is to effectively quantify T cell adhesion using flow conditions.

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Behavior

A Within-subjects Experimental Protocol to Assess the Effects of Social Input on Infant EEG
Ashley M. St. John 1, Katie Kao 1, Meia Chita-Tegmark 1, Jacqueline Liederman 1, Philip G. Grieve 2, Amanda R. Tarullo 1
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.

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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. Klein 1, Hadassah Tamir 1,2,3, Muhammad Anwar 1, Robert J. Ludwig 1, Jasmine H. Kaidbey 1, Sara B. Glickstein 4, Martha G. Welch 1,2
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.

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Immunology and Infection

Co-immunoprecipitation Assay for Studying Functional Interactions Between Receptors and Enzymes
Michael Peled 1, Marianne Strazza 2, Adam Mor 2
1Pulmonary Department, The Chaim Sheba Medical Center, 2Columbia Center for Translational Immunology, Department of Medicine, Columbia University Medical Center

Here, we present a protocol for co-immunoprecipitation and an on-bead enzymatic activity assay to simultaneously study the contribution of specific protein domains of plasma membrane receptors to both enzyme recruitment and enzyme activity.

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Medicine

Ex Vivo Corneal Organ Culture Model for Wound Healing Studies
Nileyma Castro *1, Stephanie R. Gillespie *2, Audrey M. Bernstein 1
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.

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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 He 1, Liqin Xiao 1, Huimin Yue 1, Shulin Wang 1, Yuting Sun 1, Yajun Wu 1, Liyang Wang 1,4, George Ghartey-Kwansah 1, Odell D. Jones 5, Joseph L. Bryant 6, MengMeng Xu 7, Jianjie Ma 3, Xuehon Xu 1
1National Engineering Laboratory for Resource Development of Endangered Crude Drugs in Northwest of China/CGDB, Shaanxi Normal University College of Life Sciences, 2Shaanxi Key Laboratory of Ischemic Cardiovascular Disease, Shaanxi Key Laboratory of Brain disorders, Institute of Basic & Translational Medicine, Xi'an Medical University, 3Ohio State University College of Medicine, 4Hematology-Oncology Division, Department of Medicine, Beth Israel Deaconess Medical Center, Harvard Medical School, 5University of Pennsylvania ULAR, 6University of Maryland School of Medicine, 7Columbia University Medical Center

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.

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Genetics

Embryo Injections for CRISPR-Mediated Mutagenesis in the Ant Harpegnathos saltator
Kayli Sieber 1, Maya Saar 1, Comzit Opachaloemphan 2, Matthew Gallitto 3, Huan Yang 2, Hua Yan 1,4
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.

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Medicine

Modified Technique for the Use of Neonatal Murine Hearts in the Langendorff Preparation
Matthew B. Barajas 1, Richard J. Levy 2
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.

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Biology

Assessment of Open Probability of the Mitochondrial Permeability Transition Pore in the Setting of Coenzyme Q Excess
Keren K. Griffiths 1, Aili Wang 1, Richard J. Levy 1
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.

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Neuroscience

Quantification of Immunostained Caspase-9 in Retinal Tissue
Crystal K. Colón Ortiz 1, Anna M. Potenski 2, Kendra V. Johnson 1, Claire W. Chen 1, Scott J. Snipas 3, Ying Y. Jean 1, Maria I. Avrutsky *1, Carol M. Troy *1,4,5
1Department of Pathology & Cell Biology, Vagelos College of Physicians and Surgeons, Columbia University, 2Department of Molecular Pharmacology and Therapeutics, Vagelos College of Physicians and Surgeons, Columbia University, 3NCI-designated Cancer Center, Sanford Burnham Prebys Medical Discovery Institute, 4Department of Neurology, Vagelos College of Physicians and Surgeons, Columbia University, 5The Taub Institute for Research on Alzheimer’s Disease and the Aging Brain, Vagelos College of Physicians and Surgeons, Columbia University

Presented here is a detailed immunohistochemistry protocol to identify, validate, and target functionally relevant caspases in complex tissues.

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Bioengineering

An Ex Vivo Porcine Model for Hydrodynamic Testing of Experimental Aortic Valve Procedures and Novel Medical Devices
V. Reed LaSala 1, Halil Beqaj 2, Mingze Sun 1, Sabrina Castagnini 3, Senay Ustunel 1, Elizabeth Cordoves 2, Kavya Rajesh 2, Sophia Jackman 2, David Kalfa 1,4
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.

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