Skip to main content

An Improved Method for Assessing Antigen Presentation on the Surface of Plasmodium falciparum-Infected Erythrocytes by Immuno-Electron Microscopy

  • Protocol
  • First Online:
Malaria Immunology

Abstract

Immuno-electron microscopy can detect and localize antigens in cells or tissues at a resolution of several nanometers. In the case of P. falciparum-infected erythrocytes, immuno-EM studies are frequently hampered by the electron-dense nature of the hemoglobin and access of antibodies to antigenic sites, particularly if the targeted protein is presented on the host cell surface or lies in proximity to the host cell cytoskeleton. Here, we describe an improved immuno-EM protocol that overcomes these problems. The improved signal to noise ratio and the enhanced access to antigenic sites now allows one to obtain information regarding target density and distribution and, hence, additional insights into the architecture and function of parasite-induced, or -affected, structures.

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 149.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD 199.99
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info
Hardcover Book
USD 249.99
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

Similar content being viewed by others

References

  1. Stierhof YD, Schwarz H (1989) Labeling properties of sucrose-infiltrated cryosections. Scanning Microsc Suppl 3:35–46

    CAS  PubMed  Google Scholar 

  2. Lucocq J (1994) Quantitation of gold labelling and antigens in immunolabelled ultrathin sections. J Anat 184(Pt 1):1–13

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Sanchez CP, Karathanasis C, Sanchez R, Cyrklaff M, Jäger J, Buchholz B et al (2019) Single-molecule imaging and quantification of the immune-variant adhesin VAR2CSA on knobs of Plasmodium falciparum-infected erythrocytes. Commun Biol 2:172

    Google Scholar 

  4. Quadt KA, Barfod L, Andersen D, Bruun J, Gyan B, Hassenkam T et al (2012) The density of knobs on Plasmodium falciparum-infected erythrocytes depends on developmental age and varies among isolates. PLoS One 7:e45658

    Google Scholar 

  5. Griffiths G (1993) Fine structure immunocytochemistry. Springer-Verlag, Berlin

    Book  Google Scholar 

  6. Jäger J, Schwarz US (2019) PfEMP1 distribution. https://github.com/usschwarz/PfEMP1-Distribution

    Google Scholar 

Download references

Acknowledgments

This work was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation)—Project number 240245660—SFB 1129 (M.L. and U.S.S.).

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Michael Lanzer .

Editor information

Editors and Affiliations

Rights and permissions

Reprints and permissions

Copyright information

© 2022 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

About this protocol

Check for updates. Verify currency and authenticity via CrossMark

Cite this protocol

Cyrklaff, M., Sanchez, C.P., Hanebutte, L., Jäger, J., Schwarz, U.S., Lanzer, M. (2022). An Improved Method for Assessing Antigen Presentation on the Surface of Plasmodium falciparum-Infected Erythrocytes by Immuno-Electron Microscopy. In: Jensen, A.T.R., Hviid, L. (eds) Malaria Immunology. Methods in Molecular Biology, vol 2470. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2189-9_33

Download citation

  • DOI: https://doi.org/10.1007/978-1-0716-2189-9_33

  • Published:

  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2188-2

  • Online ISBN: 978-1-0716-2189-9

  • eBook Packages: Springer Protocols

Publish with us

Policies and ethics