Botelho Roberto J. (ed.) Phagocytosis and Phagosomes: Methods and Protocols

2nd edition. — Humana Press, 2017. — 385 p. — (Methods in Molecular Biology 2692). — ISBN 978-1-0716-3337-3.This volume explores updated and entirely new experimental approaches used to investigate phagocytosis and phagosome maturation. In order to aid in the study of engulfment, maturation, resolution, and pathogen manipulation of phagocytes, the book features methodology to quantify uptake and maturation specific to certain phagocytes, particles, or pathogens, while other chapters offer methods that can be applied generically across the field. Methods are presented to study phagocytosis and phagosome maturation in vivo, in cellulo, and through in vitro analyses. Written for the highly successful Methods in Molecular Biology series, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step and readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls. Authoritative and up-to-date, Phagocytosis and Phagosomes: Methods and Protocols, Second Edition serves as a vital resource for both experts in the field as well as for investigators delving into phagocytosis and phagosome maturation for the first time.Bacterial Binding, Phagocytosis, and Killing Capacity:​ Measurements Using Colony Forming Units
Analysis of Human and Mouse Neutrophil Phagocytosis by Flow Cytometry
Quantifying Phagocytosis by Immunofluorescen​ce and Microscopy
Methods for Quantitative Efferocytosis Assays
Analysis of Efferocytic Receptor Dynamics and Synapse Formation in a Frustrated Efferocytosis Model
Super-Resolution Spinning-Disk Confocal Microscopy Using Optical Photon Reassignment (SoRa) to Visualize the Actin Cytoskeleton in acrophages
Filamentous Bacteria as Targets to Study Phagocytosis
The Derivation and Use of HoxB8-Driven Conditionally Immortalized Macrophages
Quantitative Immunofluorescen​ce to Study Phagosome Maturation and Resolution
Approaches to Measuring Reductive and Oxidative Events in Phagosomes
Measuring Phagosomal pH by Fluorescence Microscopy
Multiplexed Phagosomal Assays for the Detection and Quantification of Bidirectional Exchange Between the Phagolysosomal Lumen and Extracellular Space
Quantitative Spatio-temporal Analysis of Phagosome Maturation in Live Cells
Measurement of Salmonella enterica Internalization and Vacuole Lysis in Epithelial Cells
Microscopy-Based Tracking and Quantification Methods to Study Phagosome Resolution
Isolation of Polystyrene Bead-Induced Phagosomes for Western Blotting
Biochemically Reconstituted Fusion of Phagosomes with Endosomes and Lysosomes
An In Vitro System to Analyze Generation and Degradation of Phagosomal Phosphatidylinos​itol Phosphates
Dissecting Phagosomal Pattern Recognition Receptor-Dependent Signaling and Antigen MHC-II Presentation from Phagosomes in Murine Dendritic Cells
Examining the Kinetics of Phagocytosis-Coupled Inflammasome Activation in Murine Bone Marrow-Derived Dendritic Cells
Analysis of LC3-Associated Phagocytosis and Antigen Presentation in Macrophages and B Cells
Visualizing Phagocytic Cargo In Vivo from Engulfment to Resolution in Caenorhabditis elegans
Assessing the Phagosome Proteome by Quantitative Mass Spectrometry
Using Ion Substitution and Fluid Indicators to Monitor Macropinosome Dynamics in Live Cells