University of Connecticut

21 articles published in JoVE

• Bioengineering

  
  Fabrication and Characterization of Layer-By-Layer Janus Base Nano-Matrix to Promote Cartilage Regeneration Maxwell Landolina*¹, Anne Yau*¹, Yupeng Chen^{1 1}Department of Biomedical Engineering, University of Connecticut This protocol describes the assembly of a layer-by-layer Janus base nano-matrix (JBNm) scaffold by adding Janus base nanotubes (JBNts), matrilin-3, and Transforming Growth Factor Beta-1 (TGF-β1) sequentially. The JBNm was fabricated and characterized; additionally, it displayed excellent bioactivity, encouraging cell functions such as adhesion, proliferation, and differentiation.

• Biology

  
  Using a Thermal Camera to Measure Heat Loss Through Bird Feather Coats Jordana M.F. Graveley¹, Kevin R. Burgio¹, Margaret Rubega^{1 1}Department of Ecology and Evolutionary Biology, University of Connecticut This protocol describes the quantification of heat transmission through a flat skinned avian specimen using a thermal camera and hot water bath. The method allows obtaining of quantitative, comparative data about the thermal performance of feather coats across species using dried flat skin specimens.

• Bioengineering

  
  Fabrication of a Biomimetic Nano-Matrix with Janus Base Nanotubes and Fibronectin for Stem Cell Adhesion Libo Zhou¹, Anne Yau¹, Wuxia Zhang¹, Yupeng Chen^{1 1}Department of Biomedical Engineering, University of Connecticut The goal of this protocol is to show the assembly of a biomimetic nanomatrix (NM) with Janus base nanotubes (JBNTs) and fibronectin (FN). When co-cultured with human mesenchymal stem cells (hMSCs), the NMs exhibit excellent bioactivity in encouraging hMSCs adhesion.

• Biology

  
  A Finite Element Approach for Locating the Center of Resistance of Maxillary Teeth Bill Luu¹, Edward Anthony Cronauer², Vaibhav Gandhi¹, Jonathan Kaplan³, David M. Pierce^3,4, Madhur Upadhyay^{1 1}Division of Orthodontics, University of Connecticut Health, ²Private Practice, Miami, FL, ³Department of Biomedical Engineering, University of Connecticut, ⁴Department of Mechanical Engineering, University of Connecticut This study outlines the necessary tools for utilizing low-dose three-dimensional cone beam-based patient images of the maxilla and maxillary teeth to obtain finite element models. These patient models are then used to accurately locate the CRES of all the maxillary teeth.

• Chemistry

  
  Development and Validation of Chromium Getters for Solid Oxide Fuel Cell Power Systems Ashish Aphale¹, Junsung Hong¹, Boxun Hu^1,2, Prabhakar Singh^{1 1}Department of Materials Science and Engineering, University of Connecticut, ²Center for Clean Energy Engineering, University of Connecticut Cathode poisoning from airborne contaminants in trace levels remains a major concern for long-term stability of high-temperature electrochemical systems. We provide a novel method to mitigate the cathode degradations using getters, which capture airborne contaminants at high temperature before entering electrochemically active stack area.

• Genetics

  
  Genetic Engineering of Dictyostelium discoideum Cells Based on Selection and Growth on Bacteria Peggy Paschke¹, David A. Knecht², Thomas D. Williams¹, Peter A. Thomason³, Robert H. Insall³, Jonathan R. Chubb^4,5, Robert R. Kay¹, Douwe M. Veltman^{1 1}MRC Laboratory of Molecular Biology, ²Department of Molecular and Cell Biology, University of Connecticut, ³Cancer Research UK Beatson Institute Glasgow, ⁴MRC Laboratory for Molecular Cell Biology, University College London, ⁵Department of Cell and Developmental Biology, University College London Dictyostelium discoideum is a popular model organism to study complex cellular processes such as cell migration, endocytosis, and development. The utility of the organism is dependent on the feasibility of genetic manipulation. Here, we present methods to transfect Dictyostelium discoideum cells that overcome existing limitations of culturing cells in liquid media.

• Neuroscience

  
  A Familiarization Protocol Facilitates the Participation of Children with ASD in Electrophysiological Research Jacqueline Turcios^1,2, Barbara Cook¹, Julia Irwin^2,3, Taylor Rispoli⁴, Nicole Landi^{2,5 1}Department of Communication Sciences and Disorders, Southern Connecticut State University, ²Haskins Laboratories, ³Department of Psychology, Southern Connecticut State University, ⁴Department of Social Work, Southern Connecticut State University, ⁵Department of Psychology, University of Connecticut Electrophysiological research is an important tool for identifying biomarkers of developmental disorders, including Autism Spectrum Disorders (ASD), but data collection in these populations remains challenging. This work presents a familiarization protocol to accompany research that includes electroencephalography (EEG) to improve the likelihood of collecting EEG data from children with ASD.

• Bioengineering

  
  Magnetic Levitation Coupled with Portable Imaging and Analysis for Disease Diagnostics Stephanie M. Knowlton¹, Bekir Yenilmez², Reza Amin², Savas Tasoglu^{1,2 1}Biomedical Engineering Department, University of Connecticut, ²Mechanical Engineering Department, University of Connecticut We present a magnetic levitation technique coupled with automated imaging and analysis in both a smartphone-compatible device and a device with embedded imaging and processing. This is applied to measure the density distribution of cells with two demonstrated biomedical applications: sickle cell disease diagnosis and separating white and red blood cells.

• Biology

  
  High-Throughput, Multi-Image Cryohistology of Mineralized Tissues Nathaniel A. Dyment¹, Xi Jiang¹, Li Chen¹, Seung-Hyun Hong², Douglas J. Adams³, Cheryl Ackert-Bicknell⁴, Dong-Guk Shin², David W. Rowe^{1 1}Department 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.

• Chemistry

  
  Real-time Monitoring of Reactions Performed Using Continuous-flow Processing: The Preparation of 3-Acetylcoumarin as an Example Trevor A. Hamlin¹, Nicholas E. Leadbeater^{1 1}Department of Chemistry, University of Connecticut Real-time monitoring allows for fast optimization of reactions performed using continuous-flow processing. Here the preparation of 3-acetylcoumarin is used as an example. The apparatus for performing in-situ Raman monitoring is described, as are the steps required to optimize the reaction.

• Neuroscience

  
  3D Modeling of the Lateral Ventricles and Histological Characterization of Periventricular Tissue in Humans and Mouse Rebecca L. Acabchuk¹, Ye Sun¹, Richard Wolferz, Jr.¹, Matthew B. Eastman¹, Jessica B. Lennington¹, Brett A. Shook¹, Qian Wu², Joanne C. Conover^{1 1}Department 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.

• Neuroscience

  
  Organotypic Slice Cultures to Study Oligodendrocyte Dynamics and Myelination Robert A. Hill^1,3, Jelena Medved¹, Kiran D. Patel¹, Akiko Nishiyama^{1,2 1}Department of Physiology and Neurobiology, University of Connecticut, ²Stem Cell Institute, University of Connecticut, ³Department of Neurology, Yale University School of Medicine A technique to study NG2 cells and oligodendrocytes using a slice culture system of the forebrain and cerebellum is described. This method allows examination of the dynamics of proliferation and differentiation of cells within the oligodendrocyte lineage where the extracellular environment can be easily manipulated while maintaining tissue cytoarchitecture.

• Neuroscience

  
  The Specification of Telencephalic Glutamatergic Neurons from Human Pluripotent Stem Cells Erin M. Boisvert^1,2, Kyle Denton¹, Ling Lei¹, Xue-Jun Li^{1,3 1}Department of Neuroscience, The University of Connecticut Health Center, ²Department of Genetics and Developmental Biology, The University of Connecticut Health Center, ³Stem Cell Institute, The University of Connecticut Health Center This procedure yields telencephalic neurons by going through checkpoints which are similar to those observed during human development. The cells are allowed to spontaneously differentiate, are exposed to factors which push them towards the neural lineage, are isolated, and are plated onto coverslips to allow for terminal differentiation and maturation.

• Medicine

  
  Portable Intermodal Preferential Looking (IPL): Investigating Language Comprehension in Typically Developing Toddlers and Young Children with Autism Letitia R. Naigles¹, Andrea T. Tovar^{1 1}Department of Psychology, University of Connecticut A reliable home-based way to assess the language comprehension of very young typically developing children, as well as those with autism, is described. The method analyzes children's eye gaze while viewing side-by-side images but hearing an audio that matches only one image. Stimuli are designed with young participants in mind.

• Biology

  
  Measurement of Cellular Chemotaxis with ECIS/Taxis Kathryn M. Pietrosimone¹, Xiuyin Yin², David A. Knecht¹, Michael A. Lynes^{1 1}Molecular and Cell Biology, University of Connecticut, ²University of Connecticut The ECIS/Taxis system is an automated, real-time assay that measures cellular chemotaxis. In this assay, cells move beneath a layer of agarose to arrive at a target electrode. Cellular movement is measured by the onset of resistance to AC current 0.

• Neuroscience

  
  Autologous Blood Injection to Model Spontaneous Intracerebral Hemorrhage in Mice Lauren H. Sansing^1,2,3, Scott E. Kasner², Louise McCullough¹, Puneet Agarwal², Frank A. Welsh⁴, Katalin Kariko^{4 1}Department 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.

• Biology

  
  Insulin Injection and Hemolymph Extraction to Measure Insulin Sensitivity in Adult Drosophila melanogaster Aaron T. Haselton¹, Yih-Woei C. Fridell^{2 1}Department of Biology, State University of New York, ²Allied Health Sciences, University of Connecticut Conserved insulin signaling pathways found in the fruit fly Drosophila melanogaster make this organism a potential tool for modeling metabolic disorders including type II diabetes. To this end, it is critical to establish physiological assays to effectively measure systemic insulin action in peripheral glucose disposal in the adult fly.

• Biology

  
  Detection of Post-translational Modifications on Native Intact Nucleosomes by ELISA Bo Dai¹, Farida Dahmani², Joseph A. Cichocki³, Lindsey C. Swanson², Theodore P. Rasmussen^{3 1}Institute for Stem Cell Biology and Regenerative Medicine, Stanford University, ²Department of Molecular and Cell Biology, University of Connecticut, ³Department of Pharmaceutical Sciences, University of Connecticut Nucleosome ELISA (NU-ELISA) is a sensitive and quantitative method to detect global patterns of post-translational modifications in preparations of native, intact nucleosomes. These modifications include methylations, acetylations, and phosphorylations at specific histone amino acid residues, and hence NU-ELISA provides a global proteomic assay of the overall chromatin modification states of specific cell types.

• Neuroscience

  
  In utero Electroporation followed by Primary Neuronal Culture for Studying Gene Function in Subset of Cortical Neurons Heather Rice¹, Seiyam Suth¹, William Cavanaugh¹, Jilin Bai², Tracy L. Young-Pearse^{1 1}Center for Neurologic Diseases, Brigham and Woman's Hospital and Harvard Medical School, ²Department of Physiology and Neurobiology, University of Connecticut In utero electroporation is a valuable method for transfecting neuronal progenitor cells in vivo. Depending upon the placement of the electrodes and the developmental timepoint of electroporation, certain subsets of cortical cells can be targeted. Targeted cells can then be analyzed in vivo or in vitro for effects of genetic alteration.

• Biology

  
  Window on a Microworld: Simple Microfluidic Systems for Studying Microbial Transport in Porous Media Dmitry A. Markov^1,2, Philip C. Samson¹, David K. Schaffer¹, Adit Dhummakupt¹, John P. Wikswo^1,2,3,4, Leslie M. Shor^{5,6 1}Vanderbilt Institute for Integrative Biosystems Research and Education, Vanderbilt University, ²Department of Biomedical Engineering, Vanderbilt University, ³Department of Molecular Physiology and Biophysics, Vanderbilt University, ⁴Department of Physics and Astronomy, Vanderbilt University, ⁵Department of Chemical, Materials and Biomolecular Engineering, University of Connecticut, ⁶Center for Environmental Sciences and Engineering, University of Connecticut Microfluidic devices can be used to visualize complex natural processes in real time and at the appropriate physical scales. We have developed a simple microfluidic device that mimics key features of natural porous media for studying growth and transport of bacteria in the subsurface.

• Biology

  
  Obtaining Hemocytes from the Hawaiian Bobtail Squid Euprymna scolopes and Observing their Adherence to Symbiotic and Non-Symbiotic Bacteria Andrew J. Collins¹, Spencer V. Nyholm^{1 1}Department of Molecular and Cell Biology, University of Connecticut This video will demonstrate how to obtain hemocytes (blood cells) from the Hawaiian bobtail squid, Euprymna scolopes for use in cell biological and bacterial adhesion assays. Hemocytes will be stained with a fluorescent dye and exposed to GFP-labeled bacteria.