Skip to main content

Using Flow Cytometry to Analyze Cryptococcus Infection of Macrophages

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1519))

Abstract

Flow cytometry is a powerful analytical technique, which is increasingly being used to study the interaction between host cells and intracellular pathogens. Flow cytometry is capable of measuring a greater number of infected cells within a sample compared to alternative techniques such as fluorescence microscopy. This means that robust quantification of rare events during infection is possible. Our lab and others have developed flow cytometry methods to study interactions between host cells and intracellular pathogens, such as Cryptococcus neoformans, to quantify phagocytosis, intracellular replication, and non-lytic expulsion or “vomocytosis” from the phagosome. Herein we describe these methods and how they can be applied to the study of C. neoformans as well as other similar intracellular pathogens.

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

Buying options

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   179.99
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

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Feldmesser M, Kress Y, Novikoff P, Casadevall A (2000) Cryptococcus neoformans is a facultative intracellular pathogen in murine pulmonary infection. Infect Immun 68:4225–4237, PMC101732

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  2. Ma H, Croudace JE, Lammas DA, May RC (2006) Expulsion of live pathogenic yeast by macrophages. Curr Biol 16:2156–2160

    Article  CAS  PubMed  Google Scholar 

  3. Nicola AM, Robertson EJ, Albuquerque P, Derengowski Lda S, Casadevall A (2011) Nonlytic exocytosis of Cryptococcus neoformans from macrophages occurs in vivo and is influenced by phagosomal pH. MBio 2(4). doi:10.1128/mBio.00167,11. Print 2011

  4. Alvarez M, Casadevall A (2006) Phagosome extrusion and host-cell survival after Cryptococcus neoformans phagocytosis by macrophages. Curr Biol 16:2161–2165

    Article  CAS  PubMed  Google Scholar 

  5. Alanio A, Desnos-Ollivier M, Dromer F (2011) Dynamics of Cryptococcus neoformans-macrophage interactions reveal that fungal background influences outcome during cryptococcal meningoencephalitis in humans. MBio 2(4). doi:10.1128/mBio.00158,11. Print 2011

  6. Ma H, Hagen F, Stekel DJ, Johnston SA, Sionov E, Falk R et al (2009) The fatal fungal outbreak on Vancouver Island is characterized by enhanced intracellular parasitism driven by mitochondrial regulation. Proc Natl Acad Sci U S A 106:12980–12985, Pmc2722359

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  7. Voelz K, Johnston SA, Rutherford JC, May RC (2010) Automated analysis of cryptococcal macrophage parasitism using GFP-tagged cryptococci. PLoS One 5(12), e15968

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Robin C. May .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2017 Springer Science+Business Media New York

About this protocol

Cite this protocol

Evans, R.J., Voelz, K., Johnston, S.A., May, R.C. (2017). Using Flow Cytometry to Analyze Cryptococcus Infection of Macrophages. In: Botelho, R. (eds) Phagocytosis and Phagosomes. Methods in Molecular Biology, vol 1519. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6581-6_24

Download citation

  • DOI: https://doi.org/10.1007/978-1-4939-6581-6_24

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6579-3

  • Online ISBN: 978-1-4939-6581-6

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics