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Mansoura University

4 ARTICLES PUBLISHED IN JoVE

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.

Immunology and Infection

A Protein Microarray Assay for Serological Determination of Antigen-specific Antibody Responses Following Clostridium difficile Infection

Ola H. Negm^1,2, Mohamed Hamed^1,2, 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.

Medicine

Retinal Explant of the Adult Mouse Retina as an Ex Vivo Model for Studying Retinal Neurovascular Diseases

Khaled Elmasry^1,2,3, Mohamed Moustafa^1,2, Mohamed Al-Shabrawey^1,2

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 Aboelenain^1,2,3, Karen Schindler^1,2, Cecilia S. Blengini^1,2

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.