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Yale University School of Medicine

12 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Engineering Biological-Based Vascular Grafts Using a Pulsatile Bioreactor
Angela H. Huang 1, Laura E. Niklason 1,2
1Department of Biomedical Engineering, Yale University, 2Department of Anesthesiology, Yale University School of Medicine

Our group has developed a bioreactor culture system that mimics the physiological pulsatile stresses of the cardiovascular system to regenerate implantable small-diameter vascular grafts.

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Bioengineering

Procedure for Lung Engineering
Elizabeth A. Calle *1, Thomas H. Petersen *2, Laura E. Niklason 1,3
1Department of Biomedical Engineering, Yale University, 2Department of Biomedical Engineering, School of Medicine, Duke University, 3Department of Anesthesia, Yale University

We have developed a decellularized lung extracellular matrix and novel biomimetic bioreactor that can be used to generate functional lung tissue. By seeding cells into the matrix and culturing in the bioreactor, we generate tissue that demonstrates effective gas exchange when transplanted in vivo for short periods of time.

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Neuroscience

In vivo Laser Axotomy in C. elegans
Alexandra B. Byrne *1, Tyson J. Edwards *1, Marc Hammarlund 1
1Department of Genetics, Program in Cellular Neuroscience, Neurodegeneration and Repair, Yale University School of Medicine

A protocol to cut neurons in C. elegans with a MicroPoint pulsed laser is presented. We describe setting up the system, immobilizing worms, and severing labeled neurons. Advantages include a relatively low-cost system and the ability to sever neuronal processes or ablate cells in vivo.

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Medicine

Real-time fMRI Biofeedback Targeting the Orbitofrontal Cortex for Contamination Anxiety
Michelle Hampson 1, Teodora Stoica 1, John Saksa 2, Dustin Scheinost 1, Maolin Qiu 1, Jitendra Bhawnani 1, Christopher Pittenger 2,3,4, Xenophon Papademetris 1, Todd Constable 1
1Department of Diagnostic Radiology, Yale University School of Medicine , 2Department of Psychiatry, Yale University School of Medicine , 3Yale Child Study Center, Yale University School of Medicine , 4Interdepartmental Neuroscience Program, Yale University School of Medicine

Here we present a method for training people to control a brain area involved in contamination anxiety and for probing the relationship between contamination anxiety and brain connectivity patterns.

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Immunology and Infection

Quantitative Imaging of Lineage-specific Toll-like Receptor-mediated Signaling in Monocytes and Dendritic Cells from Small Samples of Human Blood
Feng Qian 1, Ruth R. Montgomery 1
1Department of Internal Medicine, Yale University School of Medicine

We describe use of ImageStream technology (www.amnis.com), which combines quantitative flow cytometry with simultaneous high-resolution digital imaging, to quantify cellular mechanisms of primary immune cells from well-defined patient cohorts. Our studies provide a blueprint for translational investigations to quantify lineage specific cellular responses in small samples from subject cohorts.

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Neuroscience

Preparation of Acute Subventricular Zone Slices for Calcium Imaging
Benjamin Lacar 1, Stephanie Z. Young 1, Jean-Claude Platel 1, Angélique Bordey 1
1Department of Neurosurgery and Cellular & Molecular Physiology, Yale University School of Medicine

A method to load subventricular zone (SVZ) cells with calcium indicator dyes for recording calcium activity is described. The postnatal SVZ contains tightly packed cells including neural progenitor cells and neuroblasts. Rather than using bath loading we injected the dye by pressure inside the tissue allowing better dye diffusion.

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Neuroscience

Voltage-sensitive Dye Recording from Axons, Dendrites and Dendritic Spines of Individual Neurons in Brain Slices
Marko Popovic 1, Xin Gao 1, Dejan Zecevic 1
1Department of Cellular and Molecular Physiology, Yale University School of Medicine

An imaging technique for monitoring of membrane potential changes with sub-micrometer spatial and sub-millisecond temporal resolution is described. The technique, based on laser excitation of voltage-sensitive dyes, allows measurements of signals in axons and axon collaterals, terminal dendritic branches, and individual dendritic spines.

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Biology

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization
Julie Chaumeil 1, Mariann Micsinai 1,2,3,4, Jane A. Skok 1
1Department of Pathology, New York University School of Medicine, 2New York University Center for Health Informatics and Bioinformatics, 3NYU Cancer Institute, 4Department of Pathology and Yale Cancer Center, Yale University School of Medicine

Here we describe a protocol for simultaneous detection of histone modifications by immunofluorescence and DNA sequences by DNA FISH followed by 3D microscopy and analyses (3D immuno-DNA FISH).

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Neuroscience

Neonatal Subventricular Zone Electroporation
David M. Feliciano 1, Carlos A. Lafourcade 1, Angélique Bordey 1
1Department of Neurosurgery and Cellular & Molecular Physiology, Yale University School of Medicine

We demonstrate a minimally invasive technique referred to as neonatal subventricular zone electroporation. The technique consists of injecting plasmid DNA into the lateral ventricles of neonatal pups and applying electrical current to deliver and genetically manipulate neural stem cells

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Medicine

Mouse Models for Graft Arteriosclerosis
Lingfeng Qin 1, Luyang Yu 2, Wang Min 2
1Department of Surgery, Yale University School of Medicine , 2Department of Pathology, Yale University School of Medicine

We describe protocols for our mouse graft arteriosclerois (GA) models which involve interposition of a mouse vessel segment into a recipient of the same inbred strain. By backcrossing additional genetic changes into the vessel donor, the model can assess the effect of specific genes on GA.

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JoVE Core

Imaging and Analysis of Oil Red O-Stained Whole Aorta Lesions in an Aneurysm Hyperlipidemia Mouse Model
Pei-Yu Chen 1, Lingfeng Qin 2, Michael Simons 1,3
1Yale Cardiovascular Research Center, Department of Internal Medicine, Yale University School of Medicine, 2Department of Surgery, Yale University School of Medicine, 3Department of Cell Biology, Yale University School of Medicine

This protocol provides a step-by-step procedure to analyze atherosclerotic burden in mice. Investigators can use this protocol to compare the abundance, location, and size of atherosclerotic lesions in different animals.

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Bioengineering

Engineered Lung Tissues Prepared from Decellularized Lung Slices
Katherine L. Leiby 1,2, Ronald Ng 1, Stuart G. Campbell 1,3, Laura E. Niklason 1,4
1Department of Biomedical Engineering, Yale University, 2Yale School of Medicine, 3Department of Cellular and Molecular Physiology, Yale School of Medicine, 4Department of Anesthesiology, Yale School of Medicine

This protocol describes a method to generate reproducible, small-scale engineered lung tissues, by repopulating decellularized precision-cut lung slices with alveolar epithelial type 2 cells, fibroblasts, and endothelial cells.

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