National Institutes of Health (NIH) 4 articles published in JoVE Immunology and Infection Real-Time High Throughput Technique to Quantify Neutrophil Extracellular Traps Formation in Human Neutrophils Shuichiro Nakabo1, Mariana J. Kaplan1, Sarthak Gupta1 1Systemic Autoimmunity Branch, National Institute of Arthritis and Musculoskeletal and Skin Diseases (NIAMS), National Institutes of Health (NIH) We present an automated high-throughput method to quantify neutrophil extracellular traps (NETs) utilizing the live cell analysis system, coupled with a membrane permeability-dependent dual-dye approach. Biology LipidUNet-Machine Learning-Based Method of Characterization and Quantification of Lipid Deposits Using iPSC-Derived Retinal Pigment Epithelium Zander Esh*1, Sharanya Suresh*1, Davide Ortolan1, Mitra Farnoodian1, Devika Bose1, Jiwon Ryu1, Andrei Volkov1, Kapil Bharti1, Ruchi Sharma1 1Ocular and Stem Cell Translational Research Section, National Eye Institute, National Institutes of Health (NIH) Degenerative eye diseases that affect the retinal pigment epithelium layer of the eye have monogenic and polygenic origins. Several disease models and a software application, LipidUNet, have been developed to study mechanisms of disease, as well as potential therapeutic interventions. Biology Efficient and Consistent Generation of Retinal Pigment Epithelium/Choroid Flatmounts from Human Eyes for Histological Analysis Davide Ortolan1, Andrei Volkov1, Arvydas Maminishkis1, Ruchi Sharma1, Kapil Bharti1 1Ocular and Stem Cell Translational Research Section, National Eye Institute, National Institutes of Health (NIH) We describe a method to efficiently separate retinal pigment epithelium (RPE) from the retina in human eyes and generate whole RPE/choroid flatmounts for histological and morphometric analyses of the RPE. Developmental Biology FACS-Isolation and Culture of Fibro-Adipogenic Progenitors and Muscle Stem Cells from Unperturbed and Injured Mouse Skeletal Muscle Giulia Riparini1, James M. Simone2, Vittorio Sartorelli1 1Laboratory of Muscle Stem Cells and Gene Regulation, National Institute of Arthritis, Musculoskeletal, and Skin Diseases (NIAMS), National Institutes of Health (NIH), 2Flow Cytometry Section, National Institute of Arthritis, Musculoskeletal, and Skin Diseases (NIAMS), National Institutes of Health (NIH) The precise identification of fibro-adipogenic progenitor cells (FAPs) and muscle stem cells (MuSCs) is critical to studying their biological function in physiological and pathological conditions. This protocol provides guidelines for the isolation, purification, and culture of FAPs and MuSCs from adult mouse muscles.
