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Rutgers New Jersey Medical School

9 ARTICLES PUBLISHED IN JoVE

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Biology

Using Laser Tweezers For Manipulating Isolated Neurons In Vitro
Robert Clarke 1, Jianfeng Wang 1, Ellen Townes-Anderson 1
1Department of Neurology and Neuroscience, School of Medicine, University of Medicine and Dentistry of New Jersey - UMDNJ

This video describes the manipulation of cultured neurons using laser tweezers in vitro.

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Neuroscience

Organotypic Culture of Full-thickness Adult Porcine Retina
Jianfeng Wang 1, Anton M. Kolomeyer 2, Marco A. Zarbin 2, Ellen Townes-Anderson 1
1Neurology and Neurosciences, University of Medicine and Dentistry of New Jersey - UMDNJ, 2Institute of Ophthalmology and Visual Science, University of Medicine and Dentistry of New Jersey - UMDNJ

Here we describe a cost-effective technique for organotypic culture of adult porcine retina for seven days. Briefly, a sterile filter paper was used to lift the neural retina off from the RPE and place photoreceptor side up on an insert raised by a custom-made stand.

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Neuroscience

Membrane Potential Dye Imaging of Ventromedial Hypothalamus Neurons From Adult Mice to Study Glucose Sensing
Reema P. Vazirani 1, Xavier Fioramonti 2, Vanessa H. Routh 1
1Department of Pharmacology and Physiology, Rutgers New Jersey Medical School, 2Center for Taste and Feeding Behavior, Universite de Bourgogne

The activity of single neurons from adult-aged mice can be studied by dissociating neurons from specific brain regions and using fluorescent membrane potential dye imaging. By testing responses to changes in glucose, this technique can be used to study the glucose sensitivity of adult ventromedial hypothalamic neurons.

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Medicine

An In Vitro Dormancy Model of Estrogen-sensitive Breast Cancer in the Bone Marrow: A Tool for Molecular Mechanism Studies and Hypothesis Generation
Samir Tivari 1, Reju Korah 1, Michael Lindy 1, Robert Wieder 1
1Department of Medicine and New Jersey Medical School Cancer Center, Rutgers New Jersey Medical School

We developed an in vitro model of dormancy in the bone marrow for estrogen-sensitive breast cancer cells. The goal of this protocol is to demonstrate use of the model for the study of the molecular and cellular biology of dormancy and for generation of hypotheses for subsequent testing in vivo.

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Bioengineering

Culturing Mouse Cardiac Valves in the Miniature Tissue Culture System
Boudewijn P.T. Kruithof 1, Samuel C. Lieber 2, Marianna Kruithof-de Julio 3, Vincian Gaussin 4, Marie José Goumans 1
1Department of Molecular Cell Biology, Leiden University Medical Center, 2Department of Engineering Technology, New Jersey Institute of Technology, 3Department of Urology, Leiden University Medical Center, 4Cardiovascular Research Institute, Department of Cell Biology and Molecular Medicine, Rutgers New Jersey Medical School

Here, we present an ex vivo flow model in which murine cardiac valves can be cultured allowing the study of the biology of the valve.

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Neuroscience

Immunostaining of Biocytin-filled and Processed Sections for Neurochemical Markers
Bogumila Swietek 1, Akshay Gupta 2, Archana Proddutur 2, Vijayalakshmi Santhakumar 2
1Graduate School of Biomedical Sciences, Rutgers New Jersey Medical School, 2Department of Pharmacology, Physiology and Neuroscience, Rutgers New Jersey Medical School

This protocol presents a method for the morphological recovery of neurons patched during electrophysiological recordings using biocytin filling and subsequent immunohistochemical postprocessing. We show that thick biocytin-filled sections that were stained and coverslipped can be restained with a second primary antibody days or months later.

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Bioengineering

A Versatile Method of Patterning Proteins and Cells
Anil B. Shrirao *1, Frank H. Kung *2, Derek Yip 3, Bonnie L. Firestein 2, Cheul H, Cho 3, Ellen Townes-Anderson 4
1Department of Biomedical Engineering, Rutgers University, 2Department of Cell Biology and Neuroscience, Rutgers University, 3Department of Biomedical Engineering, New Jersey Institute of Technology, 4Department of Pharmacology, Physiology, and Neuroscience, Rutgers New Jersey Medical School

This report describes a simple, easy to perform technique, using low pressure vacuum, to fill microfluidic channels with cells and substrates for biological research.

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Cancer Research

Characterization of Cell Membrane Extensions and Studying Their Roles in Cancer Cell Adhesion Dynamics
Taylor C. Brown 1, Norman G. Nicolson 2, Joyce Cheng 1, Reju Korah 1, Tobias Carling 1
1Department of Surgery & Yale Endocrine Neoplasia Laboratory, Yale University School of Medicine, 2Yale New Haven Hospital

This study demonstrates the utility and ease of quantitative cell membrane extension measurement and its correlation to adhesive capacity of cells. As a representative example, we show here that Dickkopf-related protein 3 (DKK3) promotes increased lobopodia formation and cell adhesiveness in adrenocortical carcinoma cells in vitro.

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Immunology and Infection

Intravital Imaging of Intraepithelial Lymphocytes in Murine Small Intestine
Luo Jia 1, Karen L. Edelblum 1
1Center for Immunity and Inflammation, Department of Pathology, Immunology and Laboratory Medicine, Rutgers New Jersey Medical School

We describe a method to visualize GFP-labeled γδ IELs using intravital imaging of murine small intestine by inverted spinning disk confocal microscopy. This technique enables the tracking of live cells within the mucosa for up to 4 h and can be used to investigate a variety of intestinal immune-epithelial interactions.  

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