3 articles published in JoVE
Isolation of Primary Murine Retinal Ganglion Cells (RGCs) by Flow Cytometry Sumana R. Chintalapudi1, Need N. Patel1, Zachary K. Goldsmith1, Levon Djenderedjian1, Xiang Di Wang1, Tony N. Marion2, Monica M. Jablonski1,3,4, Vanessa M. Morales-Tirado1,2 1Department of Ophthalmology, Hamilton Eye Institute, University of Tennessee Health Science Center, 2Department of Microbiology, Immunology and Biochemistry, University of Tennessee Health Science Center, 3Department of Anatomy and Neurobiology, University of Tennessee Health Science Center, 4Department of Pharmaceutical Sciences, University of Tennessee Health Science Center Millions of people suffer from retinal degenerative diseases that result in irreversible blindness. A common element of many of these diseases is the loss of retinal ganglion cells (RGCs). This detailed protocol describes the isolation of primary murine RGCs by positive and negative selection with flow cytometry.
Antibody Transfection into Neurons as a Tool to Study Disease Pathogenesis Joshua N. Douglas1,2,3, Lidia A. Gardner1,2, Sangmin Lee1,2, Yoojin Shin1,2, Chassidy J. Groover1,2, Michael C. Levin1,2,3 1Research Service, Veterans Administration Medical Center, Memphis, TN, 2Department of Neurology, University of Tennessee Health Science Center, Memphis, TN, 3Department of Anatomy/Neurobiology, University of Tennessee Health Science Center, Memphis, TN A rapid approach to investigate interactions and effects on molecular mechanisms related to the presence of antibodies in an intracellular environment is described. The method involves transfection of antibodies into live cells using a non-covalent complex formation based on a lipid formulation. The technique is adaptable to immortalized cell lines and primary cells.
Whole Cell Recording from an Organotypic Slice Preparation of Neocortex Robert C. Foehring1, Dongxu Guan1, Tara Toleman1, Angela R. Cantrell1 1Department of Anatomy and Neurobiology, University of Tennessee Health Science Center This is a protocol to prepare and maintain a neocortical slice preparation in organotypic culture for the purpose of making electrical recordings from pyramidal neurons.