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University of Connecticut Health Center

11 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice
Lauren H. Sansing 1,2,3, Scott E. Kasner 2, Louise McCullough 1, Puneet Agarwal 2, Frank A. Welsh 4, Katalin Kariko 4
1Department of Neurology, University of Connecticut Health Center, 2Department of Neurology, School of Medicine, University of Pennsylvania, 3Department of Neurosurgery, Hartford Hospital, 4Department of Neurosurgery, School of Medicine, University of Pennsylvania

The autologous blood injection model of intracerebral hemorrhage in mice described in this protocol uses the double injection technique to minimize risk of blood reflux up the needle track, no anticoagulants in the pumping system, and eliminates all dead space and expandable tubing in the system.

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Bioengineering

The Portable Chemical Sterilizer (PCS), D-FENS, and D-FEND ALL: Novel Chlorine Dioxide Decontamination Technologies for the Military
Christopher J. Doona 1, Florence E. Feeherry 1, Peter Setlow 2, Alexander J. Malkin 3, Terrence J. Leighton 4
1United States Army-Natick Soldier RD&E Center, Warfighter Directorate, 2Department of Molecular Biology and Biophysics, University of Connecticut Health Center, 3Lawrence Livermore National Laboratory, 4Children's Hospital Oakland Research Institute

The Portable Chemical Sterilizer (PCS) is a revolutionary, energy-independent, almost waterless sterilization technology for Army medical units. The PCS generates chlorine dioxide from dry reagents mixed with water on-site, at-will, and at point-of-use (PoU) in a plastic suitcase. The Disinfectant-sprayer for Foods and ENvironmentally-friendly Sanitation (D-FENS) and the Disinfectant for ENvironmentally-friendly Decontamination, All-purpose (D-FEND ALL) produce aqueous chlorine dioxide in a collapsible spray bottle and other potential embodiments. These versatile decontamination technologies kill microbes in myriad diverse Dual-use applications for military and civilian consumers.

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Neuroscience

Calcium Phosphate Transfection of Primary Hippocampal Neurons
Miao Sun *1, Laura P. Bernard *1, Victoria L. DiBona 1, Qian Wu 1, Huaye Zhang 1
1Department of Neuroscience and Cell Biology, Robert Wood Johnson Medical School, Rutgers University

Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this method on hard-to-transfect cells like primary neurons. Here we describe our detailed protocol for calcium phosphate transfection of hippocampal neurons cocultured with astroglial cells.

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Neuroscience

Determination of the Spontaneous Locomotor Activity in Drosophila melanogaster
Jared K. Woods 1, Suzanne Kowalski 1, Blanka Rogina 1
1Genetics and Developmental Biology, School of Medicine, University of Connecticut Health Center

Drosophila melanogaster are useful in studying genetic or environmental manipulations that affect behaviors such as spontaneous locomotor activity.  Here we describe a protocol that utilizes monitors with infrared beams and data analysis software to quantify spontaneous locomotor activity. 

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Neuroscience

An In Vitro Model for the Study of Cellular Pathophysiology in Globoid Cell Leukodystrophy
Kumiko I. Claycomb 1, Kasey M. Johnson 1, Ernesto R. Bongarzone 2, Stephen J. Crocker 1
1Department of Neuroscience, University of Connecticut Health Center, 2Department of Anatomy and Cell Biology, University of Illinois at Chicago

Globoid cells are a defining pathological feature of Krabbe disease, a leukodystrophy currently lacking an effective long-term therapy. We have developed a cell culture model to study the innate biology and pathogenic potential of activated microglia and their transformation into globoid cells.

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Neuroscience

3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse
Rebecca L. Acabchuk 1, Ye Sun 1, Richard Wolferz, Jr. 1, Matthew B. Eastman 1, Jessica B. Lennington 1, Brett A. Shook 1, Qian Wu 2, Joanne C. Conover 1
1Department of Physiology and Neurobiology, University of Connecticut, 2Department of Anatomic Pathology and Laboratory Medicine, University of Connecticut Health Center

Using MRI scans (human), 3D imaging software, and immunohistological analysis, we document changes to the brain’s lateral ventricles. Longitudinal 3D mapping of lateral ventricle volume changes and characterization of periventricular cellular changes that occur in the human brain due to aging or disease are then modeled in mice.

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Biology

High-Throughput, Multi-Image Cryohistology of Mineralized Tissues
Nathaniel A. Dyment 1, Xi Jiang 1, Li Chen 1, Seung-Hyun Hong 2, Douglas J. Adams 3, Cheryl Ackert-Bicknell 4, Dong-Guk Shin 2, David W. Rowe 1
1Department of Reconstructive Sciences, University of Connecticut Health Center, 2Department of Computer Science and Engineering, University of Connecticut, 3Department of Orthopaedic Surgery, University of Connecticut Health Center, 4Department of Orthopaedics, University of Rochester

In this manuscript, we present a high-throughput, semi-automated cryohistology platform to produce aligned composite images of multiple response measures from several rounds of fluorescent imaging on frozen sections of mineralized tissues.

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Biology

A Morphometric and Cellular Analysis Method for the Murine Mandibular Condyle
Eliane H. Dutra 1, Mara H. O'Brien 1, Alexandro Lima 1, Ravindra Nanda 1, Sumit Yadav 1
1Division of Orthodontics, University of Connecticut Health Center

This manuscript presents methods for analyzing morphometric and cellular changes within the mandibular condyle of rodents.

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Bioengineering

Visualization of Germinosomes and the Inner Membrane in Bacillus subtilis Spores
Juan Wen 1, Raymond Pasman 1, Erik M.M. Manders *2,3, Peter Setlow *4, Stanley Brul *1
1Molecular Biology and Microbial Food Safety, Swammerdam Institute for Life Sciences, University of Amsterdam, 2Van Leeuwenhoek Centre for Advanced Microscopy, Swammerdam Institute for Life Sciences, University of Amsterdam, 3Confocal.nl BV., 4Dept. of Molecular Biology and Biophysics, UConn Health

Germinant receptor proteins cluster in ‘germinosomes’ in the inner membrane of Bacillus subtilis spores. We describe a protocol using super resolution microscopy and fluorescent reporter proteins to visualize germinosomes. The protocol also identifies spore inner membrane domains that are preferentially stained with the membrane dye FM4-64.

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Developmental Biology

Isotropic Light-Sheet Microscopy and Automated Cell Lineage Analyses to Catalogue Caenorhabditis elegans Embryogenesis with Subcellular Resolution
Leighton H. Duncan 1,5, Mark W. Moyle 1,5, Lin Shao 1,5, Titas Sengupta 1,5, Richard Ikegami 1,5, Abhishek Kumar 4,5, Min Guo 4,5, Ryan Christensen 4,5, Anthony Santella 2,5, Zhirong Bao 2,5, Hari Shroff 4,5, William Mohler 3,5, Daniel A. Colón-Ramos 1,5,6
1Department of Neuroscience and Department of Cell Biology, Yale University School of Medicine, 2Developmental Biology Program, Sloan Kettering Institute, 3Department of Genetics and Genome Sciences and Center for Cell Analysis and Modeling, University of Connecticut Health Center, 4Section on High Resolution Optical Imaging, National Institute of Biomedical Imaging and Bioengineering, National Institutes of Health, 5WormGUIDES.org, 6Instituto de Neurobiología, Recinto de Ciencias Médicas, Universidad de Puerto Rico

Here, we present a combinatorial approach using high-resolution microscopy, computational tools, and single-cell labeling in living C. elegans embryos to understand single cell dynamics during neurodevelopment.

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Medicine

Tricolor Transgenic Murine Model for Studying Growth Plate Injury
Hui Sun 1, Natasha Patel 1, Sharif M. Ridwan 1, Christy Lottinger 1, Li Chen 1, David Rowe 1, Liisa Kuhn 1
1University of Connecticut Health Center

This protocol describes an improved mouse model for adolescent bone growth plate injuries. Using transgenic mice with tri-lineage fluorescent reporters for collagen types I, II, and X, the primary matrices associated with three different substrata of the growth plate, injury placement is guided by native fluorescence under the microscope.

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