Phagocytosis and Phagosomes: Methods and Protocols

ISBN-13: 9781493965793 / Angielski / Twarda / 2016 / 360 str.

Roberto Botelho

Opis

This volume details experimental approaches used to investigate phagocytosis and phagosome maturation. Chapters present methods and protocols on quantifying uptake and phagosome maturation using biophysical and biochemical approaches, proteomics, microscopy, and flow cytometry. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and cutting-edge, Phagocytosis and Phagosomes: Methods and Protocols aims to be an important resource for both experts in the field and for those investigators delving into phagocytosis and phagosome maturation for the first time.

Spis treści

Phagocytosis: Hungry, Hungry Cells.- Analysis of Human and Mouse Neutrophil Phagocytosis by Flow Cytometry.- Quantitative Efferocytosis Assays.- Quantifying Phagocytosis by Immunofluorescence and Microscopy.- Single Cell Analysis of Phagocytosis, Phagosome Maturation, Phagolysosomal Leakage and Cell Death Following Exposure of Macrophages to Silica Particles.- Quantitative Live-cell Fluorescence Microscopy during Phagocytosis.- Intracellular Manipulation of Phagosomal Transport and Maturation Using Magnetic Tweezers.- Quantitative Immunofluorescence to Study Phagosome Maturation.- Using Fluorescence Resonance Energy Transfer-Based Biosensors to Probe Rho Gtpase Activation during Phagocytosis.- Analysis of LC3-associated Phagocytosis and Antigen Presentation.- Quantitative Spatio-Temporal Analysis of Phagosome Maturation in Live Cells.- Measuring Phagosomal pH by Fluorescence Microscopy.- Image-Based Analysis of Phagocytosis – Measuring Engulfment and Internalization.- Fluorometric Approaches to Measuring Reductive and Oxidative Events in Phagosomes.- Simultaneous Analysis of Multiple Lumenal Parameters of Individual Phagosomes Using High-Content Imaging.- Salmonella Enterica Internalization and Vacuole Lysis in Epithelial Cells.- Bacterial Binding, Phagocytosis and Killing: Measurements Using Colony Forming Units.- Filamentous Bacteria as Targets to Study Phagocytosis.- Growing and Handling of Mycobacterium Tuberculosis for Macrophage Infection Assays.- Mycobacterium Tuberculosis: Readouts of Bacterial Fitness and The Environment within the Phagosome.- Using Flow Cytometry to Analyse Cryptococcus Infection in Macrophages.>