UConn Health 4 articles published in JoVE Immunology and Infection A Flow Cytometry-Based High-Throughput Technique for Screening Integrin-Inhibitory Drugs Ziming Cao1, Matthew J. Garcia2, Larry A. Sklar2,3,4,5, Angela Wandinger-Ness3,4, Zhichao Fan1 1Department of Immunology, School of Medicine, UConn Health, 2Center for Molecular Discovery, University of New Mexico Health Sciences Center, 3Comprehensive Cancer Center, University of New Mexico Health Sciences Center, 4Department of Pathology, University of New Mexico Health Sciences Center, 5Autophagy, Inflammation, & Metabolism (AIM) Center, University of New Mexico This protocol describes a flow cytometry-based, high-throughput screening method to identify small-molecule drugs that inhibit β2 integrin activation on human neutrophils. Immunology and Infection Real-Time Measurement of the Mitochondrial Bioenergetic Profile of Neutrophils Sunitha Pulikkot1, Meng Zhao2,3, Zhichao Fan1 1Department of Immunology, School of Medicine, UConn Health, 2Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 3Department of Microbiology and Immunology, University of Oklahoma Health Science Center We describe stepwise protocols measuring the mitochondrial respiration of mouse and human neutrophils and HL60 cells using the metabolic extracellular flux analyzer. Bioengineering Visualization of Germinosomes and the Inner Membrane in Bacillus subtilis Spores Juan Wen1, Raymond Pasman1, 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. Developmental Biology Conditional Reprogramming of Pediatric Human Esophageal Epithelial Cells for Use in Tissue Engineering and Disease Investigation Todd J. Jensen1, Christopher Foster1, Wael Sayej2, Christine M. Finck3 1Department of Pediatrics, UConn Health, 2 Expansion of human pediatric esophageal epithelial cells utilizing conditional reprogramming provides investigators with a patient-specific population of cells that can be utilized for engineering esophageal constructs for autologous implantation to treat defects or injury and serve as a reservoir for therapeutic screening assays.