Skip to main content

Visualization and Quantification of Phagocytosis by Neutrophils

  • Protocol
  • First Online:
Neutrophil

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2087))

Abstract

Phagocytosis by phagocytes such as neutrophils is a crucial part of the host innate immune response against invading pathogens. Phagocytosis is a complex process that initiates with the binding of the particles on the cell surface of the phagocytes through the interaction of pattern recognition receptors with ligands on the surface of the pathogens. During this process, phagocytes undergo extensive membrane reorganization and cytoskeleton rearrangement at their cell surface. To gain better insight about the molecular mechanisms of this dynamic cellular process, visualization and quantification in a high-throughput manner is essential. Here, we describe a microscope-based method to visualize and quantify phagocytic uptake of pathogens (such as bacteria and fungi) and model particulates that are larger than 0.5 μm (such as Zymosan A and IgG-coated beads).

This is a preview of subscription content, log in via an institution to check access.

Access this chapter

Protocol
USD 49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD 99.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 129.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 199.99
Price excludes VAT (USA)
  • Durable hardcover edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Institutional subscriptions

References

  1. Aulakh GK (2018) Neutrophils in the lung: the first responders. Cell Tissue Res 371:577–588

    Article  CAS  Google Scholar 

  2. Hauert AB, Martinelli S, Marone C, Niggli V (2002) Differentiated HL-60 cells are a valid model system for the analysis of human neutrophil migration and chemotaxis. Int J Biochem Cell Biol 34:838–854

    Article  CAS  Google Scholar 

  3. Cassatella MA (2017) Human mature neutrophils as atypical APC. Blood 129:1895–1896

    Article  CAS  Google Scholar 

  4. Lee WL, Harrison RE, Grinstein S (2003) Phagocytosis by neutrophils. Microbes Infect 5:1299–1306

    Article  CAS  Google Scholar 

  5. Koup RA, Liang F, Loré K et al (2017) Neutrophils acquire the capacity for antigen presentation to memory CD4 + T cells in vitro and ex vivo. Blood 129:1991–2001

    Article  Google Scholar 

  6. Herant M, Heinrich V, Dembo M (2006) Mechanics of neutrophil phagocytosis: experiments and quantitative models. J Cell Sci 119:1903 LP–1901913

    Article  Google Scholar 

  7. Stuart LM, Ezekowitz RAB (2005) Phagocytosis: elegant complexity. Immunity 22:539–550

    Article  CAS  Google Scholar 

  8. Fleck RA, Romero-Steiner S, Nahm MH (2005) Use of HL-60 cell line to measure opsonic capacity of pneumococcal antibodies. Clin Vaccine Immunol 12:19–27

    Article  CAS  Google Scholar 

  9. Collins SJ (1987) The HL-60 promyelocytic leukemia cell line: proliferation, differentiation, and cellular oncogene expression. Blood 70:1233–1244

    Article  CAS  Google Scholar 

  10. Tafesse FG, Rashidfarrokhi A, Schmidt FI et al (2015) Disruption of Sphingolipid biosynthesis blocks phagocytosis of Candida albicans. PLoS Pathog 11:1–27

    Article  Google Scholar 

  11. Rashidfarrokhi A, Richina V, Tafesse FG (2017) Visualizing the early stages of phagocytosis. J Vis Exp 120:54646

    Google Scholar 

  12. Polylysine-coated tissue culture surfaces. In: Protoc. https://www.protocolsonline.com/recipes/stock-solutions/polylysine-coated-tissue-culture-surfaces/. Accessed 20 May 2019

  13. Protocol for the Preparation and Fluorescent ICC Staining of Cells on Coverslips. https://www.rndsystems.com/resources/protocols/protocol-preparation-and-fluorescent-icc-staining-cells-coverslips. Accessed 20 May 2019

  14. Martin SJ, Bradley JG, Cotter TG (1990) HL-60 cells induced to differentiate towards neutrophils subsequently die via apoptosis. Clin Exp Immunol 79:448–453

    Article  CAS  Google Scholar 

  15. Millius A, Weiner OD (2010) Manipulation of neutrophil-like HL-60 cells for the study of directed cell migration. Methods Mol Biol 591:147–158

    Article  Google Scholar 

  16. Futosi K, Fodor S, Mócsai A (2013) Reprint of neutrophil cell surface receptors and their intracellular signal transduction pathways. Int Immunopharmacol 17:1185–1197

    Article  CAS  Google Scholar 

  17. Agramonte-Hevia J (2002) Gram-negative bacteria and phagocytic cell interaction mediated by complement receptor 3. FEMS Immunol Med Microbiol 34:255–266

    Article  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Fikadu G. Tafesse .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2020 Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Guzman, G., Tafesse, F.G. (2020). Visualization and Quantification of Phagocytosis by Neutrophils. In: Quinn, M., DeLeo, F. (eds) Neutrophil. Methods in Molecular Biology, vol 2087. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-0154-9_11

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-0154-9_11

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-0153-2

  • Online ISBN: 978-1-0716-0154-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics