Main description:

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

Contents:

Dedication...
Preface...
Table of Contents...
Contributing Authors...

Part I: Neutrophils and Neutrophil Disorders: Overviews

1. The Role of Neutrophils in the Immune System: An Overview
Harry L. Malech, Frank R. DeLeo, and Mark T. Quinn

2. Neutrophil Defects and Diagnosis Disorders of Neutrophil Function: An Overview
Mary C. Dinauer

Part II: Neutrophil Isolation

3. Isolation of Human Neutrophils from Venous Blood
Silvie Kremserova and William M. Nauseef

4. Isolation of Neutrophils from Non-Human Species
Daniel W. Siemsen, Lilya N. Kirpotina, Natalia Malachowa, Igor A. Schepetkin, Adeline R. Porter, Benfang Lei, Frank R. DeLeo, and Mark T. Quinn

5. Isolation of Neutrophils from Larval Zebrafish and their Transplantation into Recipient Larvae for Functional Studies
Hannah Darroch, Jonathan W. Astin, and Christopher J. Hall

Part III: Neutrophil Chemotaxis, Phagocytosis, and Bactericidal Activity

6. Analysis of Neutrophil Transmigration through Epithelial Cell Monolayers
Liliya N. Kirpotina, Douglas J. Kominsky, Mark T. Quinn, and Steve D. Swain

7. Quantification of Chemotaxis or Respiratory Burst using Ex Vivo Culture-Derived Murine Neutrophils
Klaudia Szymczak, Margery G.H. Pelletier, and Peter Gaines

8. Ex Vivo Human Neutrophil Swarming against Live Microbial Targets
Alex Hopke and Daniel Irimia

9. Micro-Injection and Micropipette Controlled Phagocytosis Methods for Neutrophils
Maurice B. Hallett, Jennie S. Campbell, Iraj Laffafian, and Sharon Dewitt

10. Using Imaging Flow Cytometry to Quantify Neutrophil Phagocytosis
Asya Smirnov, Michael D. Solga, Joanne Lannigan, and Alison K. Criss

11. Visualization and Quantification of Phagocytosis by Neutrophils
Gaelen Guzman and Fikadu G. Tafesse

12. Analysis of Neutrophil Bactericidal Activity
Nicholas J. Magon, Heather A. Parker, Louisa V. Ashby, Reuben J. Springer, and Mark B. Hampton

Part IV: Biochemistry, Biology, and Signal Transduction of Neutrophils

13. Assessment of Neutrophil Apoptosis
Nicole D. Barth, Marc Vendrell, David A. Dorward, Adriano G. Rossi, and Ian Dransfield

14. Optical Methods for the Measurement and Manipulation of Cytosolic Calcium Signals in Neutrophils
Maurice B. Hallett, Rhiannon E. Roberts, and Sharon Dewitt

15. Labeling Acidic Compartments of Neutrophils with Cresyl Violet
Philip P. Ostrowski, Ziv Roth, and Sergio Grinstein

16. Neutrophil Degranulation of Azurophil and Specific Granules
Samia Bedouhene, Pham My-Chan Dang, Margarita Hurtado-Nedelec, and Jamel El-Benna

17. Influence of Oxygen on Function and Cholesterol Composition of Murine Bone Marrow-Derived Neutrophils
Katja Branitzki-Heinemann, Graham Brogden, and Maren von Koeckritz-Blickwede

18. In Vitro Assay for Sensitive Determination of Human Blood PMN Responses
Noah Fine, William Khoury, and Michael Glogauer

19. Fast and Accurate Quantitative Analysis of Cytokine Gene Expression in Human Neutrophils by Reverse Transcription Real-Time PCR
Nicola Tamassia, Marco A. Cassatella, and Flavia Bazzoni

20. Detection of Intact Transcription Factors in Human Neutrophils
Patrick P. McDonald and Richard D. Ye

21. Genome-Scale Transcript Analyses of Human Neutrophils
Scott D. Kobayashi, Adeline R. Porter, Sarah L. Anzick, Dan E. Sturdevant, and Frank R. DeLeo

Part V: NADPH Oxidase and Production of Reactive Oxygen Species

22. Measurement of Respiratory Burst Products, Released or Retained, During Activation of Professional Phagocytes
Claes Dahlgren, Halla Bjoernsdottir, Martina Sundqvist, Karin Christenson, and Johan Bylund

23. Cell-Free NADPH Oxidase Activation Assays: A Triumph of Reductionism
Edgar Pick

Part VI: Analysis of Neutrophil Extracellular Traps

24. Immunofluorescent Detection of NET Components in Paraffin-Embedded Tissue
Ulrike Abu-Abed and Volker Brinkmann

25. Detection, Visualization, and Quantification of Neutrophil Extracellular Traps (NETs) and NET Markers
Nicole de Buhr and Maren von Koeckritz-Blickwede

26. Imaging of Neutrophils and Neutrophil Extracellular Traps (Nets) with Intravital (In Vivo) Microscopy
Iwona Cichon, Michal Santocki, Weronika Ortmann, and Elzbieta Kolaczkowska