Neutrophils, the most abundant white cells in humans, serve as the primary cellular defense against infection. This volume provides a concise set of protocols for assessing basic neutrophil functions and investigating specialized areas in neutrophil biology. Each of the protocols is written by leading researchers in the field and includes hints for success, as well as guidance for troubleshooting. In the third edition of Neutrophils: Methods and Protocols , chapters retained from previous editions have been have been updated and include many new approaches. In addition, the Third Edition contains a number of new chapters that were not included in the first or second editions. Part I contains overviews of neutrophil biology, function, and disorders. Part II describes commonly used methods to isolate neutrophils from humans and other animal species. Part III details methods for investigating chemotaxis, transmigration, phagocytosis, and bactericidal activity. Part IV includes protocols that measure neutrophil apoptosis, calcium signal transduction, degranulation and detection of cytoplasmic granules, gene expression, transcription factors, and apoptosis. Part V provides multiple assays for measuring production of intracellular and/or extracellular reactive oxygen species, and a method that details use of the cell-free NADPH oxidase assay, an iconic assay for studies of the neutrophil NADPH oxidase. Part VI provides chapters that describe how to analyze formation and function of neutrophil extracellular traps. Neutrophil Methods and Protocols: Third Edition is a comprehensive source for detailed explanations and applications of the most modern methodological advances in neutrophil biology. Both basic scientists and clinicians will find a collection of this caliber to be an invaluable aid in their work with neutrophils.

Features

• Provides neutrophil protocols that are broadly applicable to leukocyte biology

• Includes protocols for isolation of neutrophils from humans and animal species

• Presents an interesting and informative overview of the field of neutrophil biology

• Facilitates a better understanding of the role of neutrophils in host defense

Readership

Molecular and Cell Biologists, Immunologists, Infectious Disease Specialists, Microbiologists

Includes cutting-edge methods and protocols Provides step-by-step detail essential for reproducible results Contains key notes and implementation advice from the experts

Inhalt
The Role of Neutrophils in the Immune System: An Overview.- Neutrophil Defects and Diagnosis Disorders of Neutrophil Function: An Overview.- Isolation of Human Neutrophils from Venous Blood.- Isolation of Neutrophils from Non-Human Species.- Isolation of Neutrophils from Larval Zebrafish and their Transplantation into Recipient Larvae for Functional Studies.- Analysis of Neutrophil Transmigration through Epithelial Cell Monolayers.- Quantification of Chemotaxis or Respiratory Burst using Ex Vivo Culture-Derived Murine Neutrophils.- Ex Vivo Human Neutrophil Swarming against Live Microbial Targets.- Micro-Injection and Micropipette Controlled Phagocytosis Methods for Neutrophils.- Using Imaging Flow Cytometry to Quantify Neutrophil Phagocytosis.- Visualization and Quantification of Phagocytosis by Neutrophils.- Analysis of Neutrophil Bactericidal Activity.- Assessment of Neutrophil Apoptosis.- Optical Methods for the Measurement and Manipulation of Cytosolic Calcium Signals in Neutrophils.- Labeling Acidic Compartments of Neutrophils with Cresyl Violet.- Neutrophil Degranulation of Azurophil and Specific Granules.- Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils.- In Vitro Assay for Sensitive Determination of Human Blood PMN Responses.- Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils by Reverse Transcription Real-Time PCR.- Detection of Intact Transcription Factors in Human Neutrophils.- Genome-Scale Transcript Analyses of Human Neutrophils.- Measurement of Respiratory Burst Products, Released or Retained, During Activation of Professional Phagocytes.- Cell-Free NADPH Oxidase Activation Assays: A Triumph of Reductionism.- Immunofluorescent Detection of NET Components in Paraffin-Embedded Tissue.- Detection, Visualization, and Quantification of Neutrophil Extracellular Traps (NETs) and NET Markers.- Imaging of Neutrophils and Neutrophil Extracellular Traps (Nets) with Intravital (In Vivo) Microscopy.