Mansoura University 4 articles published in JoVE Medicine Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases Khaled Elmasry1,2,3, Mohamed Moustafa1,2, Mohamed Al-Shabrawey1,2 1Eye Research Center and Department of Foundational Medical Studies, Oakland University William Beaumont School of Medicine, 2Eye Research Institute, Oakland University, 3Human Anatomy and Embryology Department, Mansoura Faculty of Medicine, Mansoura University This protocol presents and describes steps for the isolation, dissection, culturing, and staining of retinal explants obtained from an adult mouse. This method is beneficial as an ex vivo model for studying different retinal neurovascular diseases such as diabetic retinopathy. Developmental Biology Evaluation of the Spindle Assembly Checkpoint Integrity in Mouse Oocytes Mansour Aboelenain1,2,3, Karen Schindler1,2, Cecilia S. Blengini1,2 1Department of Genetics, Rutgers, The State University of New Jersey, 2Human Genetics Institute of New Jersey, 3Department of Theriogenology, Faculty of Veterinary Medicine, Mansoura University Error in chromosome segregation is a common feature in oocytes. Therefore, studying the spindle assembly checkpoint gives important clues about the mechanisms needed to produce healthy eggs. The present protocol describes three complementary assays to evaluate spindle assembly checkpoint integrity in mouse oocytes. Immunology and Infection A Protein Microarray Assay for Serological Determination of Antigen-specific Antibody Responses Following Clostridium difficile Infection Ola H. Negm1,2, Mohamed Hamed1,2, Tanya M. Monaghan3 1Breast Surgery Group, Division of Medical Sciences and Graduate Entry Medicine, School of Medicine, Queen's Medical Centre, University of Nottingham, 2Medical Microbiology and Immunology Department, Faculty of Medicine, Mansoura University, 3Nottingham Digestive Diseases Centre and NIHR Nottingham Digestive Diseases Biomedical Research Centre, School of Medicine, Queen's Medical Centre, University of Nottingham This article describes a simple protein microarray method for profiling humoral immune responses to a 7-plex panel of highly purified Clostridium difficile antigens in human sera. The protocol can be extended for the determination of specific antibody responses in preparations of polyclonal intravenous immunoglobulin. Immunology and Infection Intracellular Staining and Flow Cytometry to Identify Lymphocyte Subsets within Murine Aorta, Kidney and Lymph Nodes in a Model of Hypertension Fanny Laroumanie1, Bethany L. Dale2, Mohamed A. Saleh3, Meena S. Madhur1 1Department of Medicine, Division of Clinical Pharmacology, Vanderbilt University Medical Center, 2Department of Molecular Physiology and Biophysics, Vanderbilt University, 3Department of Pharmacology and Toxicology, Faculty of Pharmacy, Mansoura University This article provides detailed methodology to identify and quantify functional T lymphocyte subsets present within murine kidney, aorta and lymph nodes by intracellular staining and flow cytometry. The model of angiotensin II induced hypertension was chosen to explain, step-by-step, the procedures and fundamental principles of flow cytometry and intracellular staining.